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:
Despite rapid clinical translation, induced pluripotent stem cell (iPSC)-derived therapies face limited global adoption. Harmonized quality control (QC) remains absent, with even fundamental parameters evaluated inconsistently across laboratories. To address this, we conducted two international Quality Assessment Rounds (QARs): QAR 2019 (18 sites, 11 countries) and QAR 2023 (23 sites, 12 countries), evaluating flow cytometry-based assessment of the undifferentiated state and qPCR-based genomic integrity testing. QAR 2019 showed high consistency in genomic integrity testing, while uncovering substantial variability in flow cytometry, prompting QAR 2023 to introduce standardized workflows. These improvements enabled systematic, cross-site evaluation of marker performance across cell states, identifying OCT3/4, TRA-1-60, and SSEA5 as consistently robust pluripotency-associated markers. This global benchmarking effort provides the first empirical multi-site evidence for reproducible iPSC QC and marker-level reliability. Together, these findings establish a foundation for harmonized QC supporting interoperable iPSC banks, regulatory alignment, and scalable manufacturing of globally accessible regenerative therapies. Keywords: induced pluripotent stem cells, regenerative medicine, cell therapy manufacturing, quality control, genomic integrity, flow cytometry, qPCR, inter-laboratory reproducibility, standardization, global harmonization

Candidates:
A. To address this, we conducted two international Quality Assessment Rounds (QARs): QAR 2020 (18 sites, 11 countries) and QAR 2023 (23 sites, 12 countries), evaluating flow cytometry-based assessment of the undifferentiated state and qPCR-based genomic integrity testing.
B. QAR 2019 showed high consistency in genomic integrity testing, while uncovering substantial variability in flow cytometry, prompting QAR 2023 to introduce standardized workflows.
C. Despite rapid clinical translation, induced pluripotent stem cell (iPSC)-derived therapies face limited global adoption.
D. The evidence does not state that despite rapid clinical translation, induced pluripotent stem cell (iPSC)-derived therapies face limited global adoption.
E. To address this, we conducted two international Quality Assessment Rounds (QARs): QAR 2019 (18 sites, 11 countries) and QAR 2023 (23 sites, 12 countries), evaluating flow cytometry-based assessment of the undifferentiated state and qPCR-based genomic integrity testing.
F. Harmonized quality control (QC) remains absent, with even fundamental parameters evaluated inconsistently across laboratories.
G. QAR 2020 showed high consistency in genomic integrity testing, while uncovering substantial variability in flow cytometry, prompting QAR 2023 to introduce standardized workflows.
H. The evidence does not state that harmonized quality control (QC) remains absent, with even fundamental parameters evaluated inconsistently across laboratories.


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:
Human induced pluripotent stem cells (hiPSCs) possess broad differentiation potential; however, efficient maturation into functional hepatocytes remains challenging, in part because conventional differentiation strategies rely on 2D culture or partially defined 3D systems that fail to recapitulate key developmental microenvironmental cues. Although 3D culture has been shown to promote hepatic specification and maturation, a fully defined platform that supports the entire differentiation process from hiPSCs in 3D has not been established. Here, we report a scalable synthetic peptide hydrogel (PepGel) platform that enables hiPSCs to self-organize into long-term, proliferative luminal cavity (LC) architectures within 3D colonies (LC-hiPSCs). These LC-hiPSCs exhibited markedly enhanced differentiation efficiency compared to hiPSC aggregates generated via scaffold-free suspension methods. Using a shear-thinning, self-healing hydrogel configuration optimized for suspension culture, we produced large quantities of LC-hiPSCs and directed their differentiation into PepGel-derived hiPSC-derived hepatocyte-like cells (PG-hiHs). PG-hiHs formed polarized, multi-luminal organoid-like structures and exhibited robust expression of hepatocyte-specific genes and proteins, high cytochrome P450 activity, and in vitro metabolic functions comparable to primary human hepatocytes (PHHs). Following live shipment and transplantation into immunocompromised mouse livers, PG-hiHs engrafted efficiently and maintained human-specific albumin and alpha-1 antitrypsin secretion at PHH-equivalent levels. Furthermore, both LC-hiPSCs and PG-hiHs retained viability, structural organization, and phenotype in complex 3D bioprinted constructs for at least 14 days. Together, this work establishes a fully defined, reproducible, and scalable 3D platform that supports the entire differentiation process, overcoming limitations in hiPSC-derived hepatic maturation and enables physiologically relevant hepatocyte-like tissues for disease modeling, regenerative medicine, and biofabrication.

Candidates:
A. Human induced pluripotent stem cells (hiPSCs) possess broad differentiation potential; however, efficient maturation into functional hepatocytes remains challenging, in part because conventional differentiation strategies rely on 3D culture or partially defined 3D systems that fail to recapitulate key developmental microenvironmental cues.
B. Human induced pluripotent stem cells (hiPSCs) possess broad differentiation potential; however, efficient maturation into functional hepatocytes remains challenging, in part because conventional differentiation strategies rely on 2D culture or partially defined 3D systems that fail to recapitulate key developmental microenvironmental cues.
C. Although 4D culture has been shown to promote hepatic specification and maturation, a fully defined platform that supports the entire differentiation process from hiPSCs in 3D has not been established.
D. Here, we report a scalable synthetic peptide hydrogel (PepGel) platform that enables hiPSCs to self-organize into long-term, proliferative luminal cavity (LC) architectures within 4D colonies (LC-hiPSCs).
E. Here, we report a scalable synthetic peptide hydrogel (PepGel) platform that enables hiPSCs to self-organize into long-term, proliferative luminal cavity (LC) architectures within 3D colonies (LC-hiPSCs).
F. Although 3D culture has been shown to promote hepatic specification and maturation, a fully defined platform that supports the entire differentiation process from hiPSCs in 3D has not been established.
G. The evidence does not state that these LC-hiPSCs exhibited markedly enhanced differentiation efficiency compared to hiPSC aggregates generated via scaffold-free suspension methods.
H. These LC-hiPSCs exhibited markedly enhanced differentiation efficiency compared to hiPSC aggregates generated via scaffold-free suspension 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:
New enzymatic sources need to be explored to meet the increasing demand for bioactives used in the supplementation of non-ruminant animals. Phytase and xylanase are widely used in animal nutrition due to their ability to degrade phytic acid and xylan, respectively. This study aimed to (i) produce phytase and xylanase by culturing Aspergillus japonicus on medium containing wheat bran + soybean bran (1:1 w/w) as alternative carbon sources in a stirred-tank bioreactor and (ii) compare this process with that conducted in Erlenmeyer flasks. The resulting enzymatic cocktails and commercial enzymes were evaluated for their ability to hydrolyze poultry diets (D1–D9). The production peaks of phytase and xylanase in the bioreactor occurred at 120 h (842.30 ± 43.62 U/L) and 144 h (3211.29 ± 274.98 U/L), respectively. After 144 h of cultivation, protein concentration in the bioreactor was approximately twice that observed in Erlenmeyer flasks. ECPAjBr displayed the highest hydrolytic activity for inorganic phosphate in diet D6 (42.34 ± 2.20 mg/g), followed by diet D7 (38.67 ± 0.46 mg/g). Regarding reducing sugars, ECXAjBr exhibited the highest activity in diets D1 and D8, releasing 147.71 ± 18.18 mg/g and 145.83 ± 11.39 mg/g, respectively. All tested diets, regardless of their composition, indicated a synergistic interaction between commercial enzymes and those present in the A. japonicus enzyme cocktail, which exhibits a broad spectrum of hydrolytic activities, including high levels of accessory enzymes. The application of this cocktail in animal diets holds great potential for increasing nutrient bioavailability in non-ruminant animal production.

Candidates:
A. The evidence does not state that new enzymatic sources need to be explored to meet the increasing demand for bioactives used in the supplementation of non-ruminant animals.
B. Phytase and xylanase are not widely used in animal nutrition due to their ability to degrade phytic acid and xylan, respectively.
C. New enzymatic sources need to be explored to meet the increasing demand for bioactives used in the supplementation of non-ruminant animals.
D. The resulting enzymatic cocktails and commercial enzymes were evaluated for their ability to hydrolyze poultry diets (D2–D9).
E. This study aimed to (i) produce phytase and xylanase by culturing Aspergillus japonicus on medium containing wheat bran + soybean bran (2:1 w/w) as alternative carbon sources in a stirred-tank bioreactor and (ii) compare this process with that conducted in Erlenmeyer flasks.
F. The resulting enzymatic cocktails and commercial enzymes were evaluated for their ability to hydrolyze poultry diets (D1–D9).
G. Phytase and xylanase are widely used in animal nutrition due to their ability to degrade phytic acid and xylan, respectively.
H. This study aimed to (i) produce phytase and xylanase by culturing Aspergillus japonicus on medium containing wheat bran + soybean bran (1:1 w/w) as alternative carbon sources in a stirred-tank bioreactor and (ii) compare this process with that conducted in Erlenmeyer flasks.


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:
Duckweeds ( Lemnaceae ), the smallest and fastest-growing flowering plants, have emerged as a transformative platform for sustainable biotechnology. This review synthesizes recent advances that underpin their potential as a next-generation plant chassis. We discuss duckweed's unique biology, characterized by reductive evolution, extreme phenotypic plasticity, and a simplified epigenome that favors transgene expression. The decoding of its minimalist genome, along with the establishment of efficient genetic tools including optimized transformation and CRISPR-Cas9 editing, enables precise genetic and metabolic engineering. While traditional uses in phytoremediation and animal feed validate its utility, duckweed's rapid growth in contained, soil-free culture and its edibility offer distinct advantages for molecular farming over established systems like tobacco. We highlight progress in engineering duckweeds to produce vaccines, therapeutic proteins, and high-value metabolites. To transition from proof-of-concept to an industrial workhorse, future efforts must focus on integrated omics databases, universal genetic toolkits, and scalable cultivation. Converging fundamental insights with synthetic biology principles positions duckweed as a versatile and powerful chassis for the bioeconomy.

Candidates:
A. This review synthesizes recent advances that underpin their potential as a next-generation plant chassis.
B. We discuss duckweed's unique biology, characterized by reductive evolution, extreme phenotypic plasticity, and a simplified epigenome that favors transgene expression.
C. The decoding of its minimalist genome, along with the establishment of efficient genetic tools including optimized transformation and CRISPR-Cas9 editing, enables precise genetic and metabolic engineering.
D. The evidence does not state that this review synthesizes recent advances that underpin their potential as a next-generation plant chassis.
E. The decoding of its minimalist genome, along with the establishment of efficient genetic tools including optimized transformation and CRISPR-Cas10 editing, enables precise genetic and metabolic engineering.
F. Duckweeds ( Lemnaceae ), the smallest and fastest-growing flowering plants, have emerged as a transformative platform for sustainable biotechnology.
G. The evidence does not state that we discuss duckweed's unique biology, characterized by reductive evolution, extreme phenotypic plasticity, and a simplified epigenome that favors transgene expression.
H. The evidence does not state that duckweeds ( Lemnaceae ), the smallest and fastest-growing flowering plants, have emerged as a transformative platform for sustainable 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:
Optical coherence tomography velocimetry (OCTV) was demonstrated with in-line processing of biologics for the first time. OCTV allowed the velocity of concentrated monoclonal antibodies (mAbs at 39.5–84.7 mg mL –1 ) to be probed in 3.4 pL volumes over distances 0–5 mm from the pipe walls. The large penetration depth is facilitated by the relatively low turbidity of mAbs at near-infrared wavelengths (1300 nm). The mAb solutions could be concentrated in situ and the changes to the viscoelasticity measured. Higher concentration mAb solutions became shear thinning (following the power law fluid model) and the amplitude of their velocity fluctuations decreased. Furthermore, dropping the pH of the mAb solutions induced a gelation phase transition and complex changes to the mAb rheology could be observed with OCTV e.g. thixotropy and the formation of a stationary boundary layer. Thus, in situ formulation of mAbs could be explored with OCTV under industrially relevant conditions.

Candidates:
A. The large penetration depth is facilitated by the relatively low turbidity of mAbs at near-infrared wavelengths (1300 nm).
B. The mAb solutions could be concentrated in situ and the changes to the viscoelasticity measured.
C. The evidence does not state that the mAb solutions could be concentrated in situ and the changes to the viscoelasticity measured.
D. Optical coherence tomography velocimetry (OCTV) was demonstrated with in-line processing of biologics for the first time.
E. OCTV allowed the velocity of concentrated monoclonal antibodies (mAbs at 40.5–84.7 mg mL –1 ) to be probed in 3.4 pL volumes over distances 0–5 mm from the pipe walls.
F. The large penetration depth is facilitated by the relatively low turbidity of mAbs at near-infrared wavelengths (1301 nm).
G. OCTV allowed the velocity of concentrated monoclonal antibodies (mAbs at 39.5–84.7 mg mL –1 ) to be probed in 3.4 pL volumes over distances 0–5 mm from the pipe walls.
H. Optical coherence tomography velocimetry (OCTV) was not demonstrated with in-line processing of biologics for the first time.


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:
Antimicrobial peptides (AMPs) such as pediocin PA-1 are attractive for food biopreservation and infection control, but their broader use is limited by low recombinant yields and high production costs. Corynebacterium glutamicum has emerged as a robust GRAS chassis for heterologous peptide and protein production, yet commonly used shuttle vectors provide only moderate plasmid copy numbers and expression capacities. In particular, existing pediocin PA-1 processes in C. glutamicum rely on standard pBL1 - or pCG1-family vectors that do not yet leverage replication-origin engineering. We rationally redesigned the replication control region of the widely used pClik 5α (pCG1-family) backbone by introducing targeted mutations in the repA gene, an antisense RNA ( cgrI ) promoter, and putative partitioning genes parAB , and constructed a systematic panel of high-copy variants. Using a P tuf -driven mCherry reporter as a quantitative readout, we identified plasmids that supported several-fold higher fluorescence than the parental backbone while maintaining robust growth. Fluorescence-based gene-dosage estimation indicated a strong increase in apparent plasmid copy number. Independent qPCR-based plasmid copy number determination using two plasmid loci confirmed that the lead variant pClik 5α repA mut reached approximately 28–30 copies per chromosome equivalent, compared to approximately 2–3 copies for the parental plasmid, corresponding to an approximately 10-fold increase. Genome-wide transcriptome analysis revealed a defined and adaptive transcriptional response to elevated plasmid copy number and expression burden, characterized by adjustments in membrane-associated transport, respiratory functions, and amino acid-related metabolism, without evidence of collapse of core biosynthetic functions. When the best-performing replicon was applied to episomal expression of a codon-optimized pedACD Cgl operon, pediocin PA-1 titers increased by 2.5-fold compared to the best pXMJ19-based reference under identical, previously optimized process conditions, placing the system, under comparable cultivation formats, within the upper range of reported microbial pediocin production processes. This work demonstrates that rational engineering of pCG1-family replication modules in C. glutamicum can unlock markedly higher plasmid copy numbers and expression capacities while preserving physiological robustness. The resulting high-copy pClik 5α derivatives, exemplified by pClik 5α repA mut , provide a versatile high-copy expression platform with demonstrated utility for recombinant reporter protein and antimicrobial peptide production in C. glutamicum and offer a foundation for further integration with folding, secretion, and process engineering strategies to advance industrial AMP production. The online version contains supplementary material available at 10.1186/s12934-026-03004-y. Keywords: Corynebacterium glutamicum , Pediocin PA-1, Bioprocess, Food additive, Bacteriocin, Antimicrobial peptide, Plasmid copy number, Expression system, Peptide production, mCherry, Listeria spp


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:
Escherichia coli is a microbial expression system widely spread in industrial manufacturing systems for the production of biotherapeutics. Most of these recombinant protein production processes are established in fed-batch operation mode. While many sectors of the chemical industry have implemented intensification strategies, the bio-pharmaceutical sector lacks a profound change towards intensified operations due to a decrease of cellular productivity during extended cultivation periods. In this study, intensified bioreactor cultivations of an auto-inducible E. coli strain W3110 as a promising candidate for the production of a recombinant fragment antigen binding (Fab) are demonstrated. The main goal was to investigate the impact of the natural phosphate limitation on the activity of the alkaline phosphatase promoter (phoA) in terms of cell-specific productivity in different processing modes. Following a cell-physiological characterization in fed-batch operation, the transition to intensified upstream processing modes, such as repetitive fed-batch and chemostat was established successfully. A comprehensive understanding of phosphate limitation and precise control of critical parameters, such as growth rate and phosphate-to-substrate uptake enabled the intensified process to match specific product titers and surpass process efficiency (in terms of space-time yield) compared to fed-batch mode. The auto-inducible phoA promoter in E. coli W3110 enabled stable Fab production under intensified bioprocess operations. These findings highlight the suitability of this expression system for repetitive fed-batch and chemostat operations to support broader adoption of intensified strategies in the bio-pharmaceutical industry. The online version contains supplementary material available at 10.1186/s12866-026-04887-y. Keywords: Intensified biomanufacturing, Escherichia coli , phoA expression system, Recombinant protein production, Fragment antigen binding

Candidates:
A. In this study, intensified bioreactor cultivations of an auto-inducible E.
B. The evidence does not state that in this study, intensified bioreactor cultivations of an auto-inducible E.
C. Escherichia coli is a microbial expression system widely spread in industrial manufacturing systems for the production of biotherapeutics.
D. The evidence does not state that while many sectors of the chemical industry have implemented intensification strategies, the bio-pharmaceutical sector lacks a profound change towards intensified operations due to a decrease of cellular productivity during extended cultivation periods.
E. Most of these recombinant protein production processes are not established in fed-batch operation mode.
F. While many sectors of the chemical industry have implemented intensification strategies, the bio-pharmaceutical sector lacks a profound change towards intensified operations due to a decrease of cellular productivity during extended cultivation periods.
G. Escherichia coli is not a microbial expression system widely spread in industrial manufacturing systems for the production of biotherapeutics.
H. Most of these recombinant protein production processes are established in fed-batch operation mode.


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: ["1.78 MPa", "0.78 MPa"]

Evidence:
Tendon gel is a translucent gel-like material secreted from the ends of a severed tendon. When mechanical stress is applied to a 3-day in vivo-preserved tendon gel, it matures into type I collagen–dominant tissue similar to normal tendon. This study aimed to evaluate the effects of transplanting a 3-day in vivo-preserved tendon gel into a knee medial collateral ligament (MCL) injury site in rabbits to promote intrinsic ligament regeneration. Tendon gel was prepared from rabbit Achilles tendons using the film model method and harvested after 3 days of in vivo preservation. The 3-day tendon gel was transplanted into the knee MCL injury sites in another set of rabbits ( n = 48). Additionally, the healing process was assessed at 1, 2, and 4 weeks postoperatively using mechanical and histological analyses. Ultimate load, peak stress, and elastic modulus were measured. Histological maturity was semi-quantitatively scored, and collagen type I and III expressions were examined by immunofluorescence staining. At 2 weeks, the tendon gel group demonstrated significantly higher ultimate load than the control group (12.25 ± 4.90 vs. 5.25 ± 2.40 N; p = 0.02). The tendon gel group had greater peak stress than the control group (3.54 ± 1.44 vs. 1.69 ± 0.78 MPa; p = 0.02). Histological scores were higher in the tendon gel group than in the control group (7.25 ± 0.43 vs. 5.50 ± 1.73; p = 0.03). Cells in the tendon gel group were aligned parallel to collagen fibers with elongated nuclei, while type I collagen expression was stronger than that observed in controls. Transplanting a 3-day in vivo-preserved tendon gel into an injured ligament enhanced mechanical strength and histological maturation at 2 weeks postoperatively. These findings suggest that this tendon gel serves as a promising biomaterial for accelerating ligament healing.


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:
Sourdough fermentation is a traditional biotechnological process used to improve the nutritional value, flavour profile, and textural attributes of baked goods. Sourdough principal microflora is represented by yeasts, lactic acid bacteria (LAB) and acetic acid bacteria (AAB). The yeasts are primarily responsible for leavening whereas the LABs and AABs carry out the acidification of the dough and both contribute to the flavour of the resulting bread. Our previous work explored the microbial species diversity of sourdoughs in Botswana and showed the unique microbial communities involved in sourdough production from different areas in Botswana. This study is aimed at characterizing yeasts and bacterial isolates from 9 traditional sourdoughs indigenous to Botswana. Here we report the functional characterization of 21 yeasts, 9 LAB, 4 AAB, and 11 other bacteria isolated from traditional sourdough as reported in our previous study. These isolates were characterized based on their ability to assimilate different carbon sources found in flour (maltose, glucose, sucrose, fructose, raffinose and maltotriose), capability to ferment carbon sources (maltose, glucose, sucrose and fructose) in flour and ability to withstand baking associated stresses (thermal stress, osmotic stress, oxidative stress and ethanol stress). This characterization aimed to compare them with conventional baker’s yeast used in modern bread making and to assess their potential for future commercialization. Our results show that the majority of isolated S. cerevisiae strains can utilise different carbon sources, ferment them and withstand baking associated stresses. These are key baking traits poorly expressed in the conventional baker’s yeast. In contrast, the isolated LAB and AAB exhibited limited carbon source utilization. Despite the noted poor utilization of these isolates, we noticed that L. plantarum (ME1-B1), B. cereus (GT1-B1) and B. zhangzhouensis were able to withstand baking associated stresses better than all other bacteria isolates. This study shows how isolated S. cerevisiae strains and non-conventional yeasts have comparable capability to the conventional baker’s yeast, highlighting their potential as sourdough starters and prospects for future commercialization. The online version contains supplementary material available at 10.1007/s00284-026-04880-8.

Candidates:
A. The yeasts are not primarily responsible for leavening whereas the LABs and AABs carry out the acidification of the dough and both contribute to the flavour of the resulting bread.
B. The evidence does not state that our previous work explored the microbial species diversity of sourdoughs in Botswana and showed the unique microbial communities involved in sourdough production from different areas in Botswana.
C. Sourdough principal microflora is represented by yeasts, lactic acid bacteria (LAB) and acetic acid bacteria (AAB).
D. Our previous work explored the microbial species diversity of sourdoughs in Botswana and showed the unique microbial communities involved in sourdough production from different areas in Botswana.
E. Sourdough fermentation is not a traditional biotechnological process used to improve the nutritional value, flavour profile, and textural attributes of baked goods.
F. Sourdough fermentation is a traditional biotechnological process used to improve the nutritional value, flavour profile, and textural attributes of baked goods.
G. The yeasts are primarily responsible for leavening whereas the LABs and AABs carry out the acidification of the dough and both contribute to the flavour of the resulting bread.
H. Sourdough principal microflora is not represented by yeasts, lactic acid bacteria (LAB) and acetic acid bacteria (AAB).


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:
Growing global protein demand has fueled innovation and investment in alternative protein (AP) products, including plant-based, fermentation-derived, and cell-cultivated products. Through interviews with AP stakeholders in the United States (U.S.), we explored the sector’s evolution, challenges, opportunities, and trends. Interviewees described a boom from 2009 to 2021 followed by a decline, which the sector is now working to reverse. Achieving taste and price parity, attracting a broad consumer base, producing at scale, and navigating a charged policy environment remain key sector challenges. Looking forward, stakeholders were optimistic, noting opportunities including collaborations within and across sectors; workforce development; innovative financing, scalability models, and products; and increased policy engagement. Findings indicate that the U.S. AP sector is at a critical inflection point. This study points to future research, financing, and innovation that could help AP products become a mainstay in the food system. Subject terms: Business and management, Business and management, Economics, Economics, Environmental social sciences, Information systems and information technology

Candidates:
A. The evidence does not state that through interviews with AP stakeholders in the United States (U.S.), we explored the sector’s evolution, challenges, opportunities, and trends.
B. The evidence does not state that achieving taste and price parity, attracting a broad consumer base, producing at scale, and navigating a charged policy environment remain key sector challenges.
C. Growing global protein demand has fueled innovation and investment in alternative protein (AP) products, including plant-based, fermentation-derived, and cell-cultivated products.
D. Interviewees described a boom from 2010 to 2021 followed by a decline, which the sector is now working to reverse.
E. Interviewees described a boom from 2009 to 2021 followed by a decline, which the sector is now working to reverse.
F. Through interviews with AP stakeholders in the United States (U.S.), we explored the sector’s evolution, challenges, opportunities, and trends.
G. The evidence does not state that growing global protein demand has fueled innovation and investment in alternative protein (AP) products, including plant-based, fermentation-derived, and cell-cultivated products.
H. Achieving taste and price parity, attracting a broad consumer base, producing at scale, and navigating a charged policy environment remain key sector challenges.


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:
As chimeric antigen receptor (CAR)‐T cell therapy has expanded rapidly to meet the growing global cancer burden; many challenges have emerged as a critical factor influencing its efficacy. However, due to the complicated mechanisms of CAR‐T cells, human interference alone was insufficient to optimize the outcomes. In parallel, artificial intelligence (AI) has begun to intersect with CAR‐T cells, offering novel computational interferences that can refine therapeutic mechanisms. The literature is still lacking a comprehensive investigation that merges CAR‐T cell mechanistic biology and limitations with the advancing abilities of AI to meet these barriers. This review provides an overview of the mechanistic foundations of CAR‐T cell. It also investigates the various challenges facing the current CAR‐T therapies including toxicity, resistance, and accessibility issues. On this basis, we examined the way AI‐based innovations are being utilized to optimize the CAR‐T engineering and clinical management. Finally, we examined clinical studies and case studies incorporating AI elements, emphasizing both therapeutic mechanisms and outcomes of the study. By integrating mechanistic biology with computational innovation, this review provides a unified unique perspective that can guide the development of safer and more effective CAR‐T therapies.

Candidates:
A. As chimeric antigen receptor (CAR)‐T cell therapy has expanded rapidly to meet the growing global cancer burden; many challenges have emerged as a critical factor influencing its efficacy.
B. However, due to the complicated mechanisms of CAR‐T cells, human interference alone was not insufficient to optimize the outcomes.
C. The literature is not still lacking a comprehensive investigation that merges CAR‐T cell mechanistic biology and limitations with the advancing abilities of AI to meet these barriers.
D. In parallel, artificial intelligence (AI) has begun to intersect with CAR‐T cells, offering novel computational interferences that can refine therapeutic mechanisms.
E. The evidence does not state that as chimeric antigen receptor (CAR)‐T cell therapy has expanded rapidly to meet the growing global cancer burden; many challenges have emerged as a critical factor influencing its efficacy.
F. In parallel, artificial intelligence (AI) has begun to intersect with CAR‐T cells, offering novel computational interferences that cannot refine therapeutic mechanisms.
G. The literature is still lacking a comprehensive investigation that merges CAR‐T cell mechanistic biology and limitations with the advancing abilities of AI to meet these barriers.
H. However, due to the complicated mechanisms of CAR‐T cells, human interference alone was insufficient to optimize the outcomes.


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:
Pseudouridine, the C5-ribose epimer of uridine with significant biological functions and clinical applications, was efficiently produced through systematic metabolic strategy of Escherichia coli in this study. Initial overexpression of pseudouridine-5-phosphate glycosylase gene psuG and alkaline phosphatase gene YjjG in E . coli pRSFDuet-1- YjjG - psuG yielded 0.43 g L −1 pseudouridine, which increased 8.56-fold with 5 g L −1 uridine supplementation. Subsequent deletion of thrA , psuT , argF , and pepA enhanced titer by 1.29-fold in E . coli Δ thrA Δ psuT Δ argF Δ pepA pRSFDuet-1- YjjG - psuG , while ribonucleoside hydrolase gene rihA overexpression boosted titer to 5.57 g L −1 . Further optimization through deleting the uridine kinase gene udk and overexpressing the ribokinase gene rbsK in strain E . coli Δ thrA Δ psuT Δ argF Δ pepA Δ udk Δ udp Δ ppnp pRSFDuet-1- YjjG - psuG pCDFDuet-1- rihA-rbsK achieved 6.23 g L −1 pseudouridine, increasing to 11.34 g L −1 with two-stage uridine feeding. Fed-batch fermentation in a 5-L bioreactor yielded a record 102.2 g L −1 pseudouridine. This work provides an efficient and scalable bioprocess for industrial pseudouridine manufacturing to meet the growing demands of mRNA-based applications. Keywords: Rational reconstruction, Pathway optimization, Enhanced production, Escherichia coli , Pseudouridine

Candidates:
A. coli pRSFDuet-1- YjjG - psuG yielded 0.43 g L −1 pseudouridine, which increased 8.56-fold with 5 g L −1 uridine supplementation.
B. Initial overexpression of pseudouridine-5-phosphate glycosylase gene psuG and alkaline phosphatase gene YjjG in E .
C. Pseudouridine, the C6-ribose epimer of uridine with significant biological functions and clinical applications, was efficiently produced through systematic metabolic strategy of Escherichia coli in this study.
D. Subsequent deletion of thrA , psuT , argF , and pepA enhanced titer by 1.29-fold in E .
E. Initial overexpression of pseudouridine-6-phosphate glycosylase gene psuG and alkaline phosphatase gene YjjG in E .
F. Pseudouridine, the C5-ribose epimer of uridine with significant biological functions and clinical applications, was efficiently produced through systematic metabolic strategy of Escherichia coli in this study.
G. Subsequent deletion of thrA , psuT , argF , and pepA enhanced titer by 2.29-fold in E .
H. coli pRSFDuet-2- YjjG - psuG yielded 0.43 g L −1 pseudouridine, which increased 8.56-fold with 5 g L −1 uridine supplementation.


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:
Chimeric antigen receptor (CAR) T cells present a novel and transformative approach to treat certain haematological malignancies. However, CAR‐T cell expansion methods are still under development and often rely on manual, low‐control culture methods. The transition to bioreactors would allow for greater process control and scalability and is a key focus of research in the field. Despite this, there are few methods to determine culture progression without the need for manual cell sampling which risks introducing errors and heightening contamination risks. In this article, we assessed whether capacitance technology could deliver reliable, on‐line cell concentrations by comparing T cell, and CAR‐T cell bioprocesses with Chinese hamster ovary (CHO) cultures—widely used in biotechnology and with established capacitance usage. Finding that capacitance technology could accurately measure CAR‐T cell concentrations, we then demonstrated the automation of feeding using capacitance‐derived triggers which improved bioprocess performance in terms of cell concentration and throughput. We anticipate that this study will expand avenues of investigation regarding capacitance as a suitable process analytical technology (PAT) to enable monitoring and control of CAR‐T cell manufacture, and potentially other cell and gene therapy products. It may also enable remote monitoring of multiple batches, harvest control, and the generation of large collections of process data for modelling, which will further progress the field. Keywords: bioprocess 4.0, cell and gene therapy, chimeric antigen receptor T cells, dielectric spectroscopy, Jurkat, process analytical technology

Candidates:
A. Chimeric antigen receptor (CAR) T cells present a novel and transformative approach to treat certain haematological malignancies.
B. However, CAR‐T cell expansion methods are not still under development and often rely on manual, low‐control culture methods.
C. The transition to bioreactors would allow for greater process control and scalability and is a key focus of research in the field.
D. The transition to bioreactors would allow for greater process control and scalability and is not a key focus of research in the field.
E. The evidence does not state that chimeric antigen receptor (CAR) T cells present a novel and transformative approach to treat certain haematological malignancies.
F. Despite this, there are not few methods to determine culture progression without the need for manual cell sampling which risks introducing errors and heightening contamination risks.
G. However, CAR‐T cell expansion methods are still under development and often rely on manual, low‐control culture methods.
H. Despite this, there are few methods to determine culture progression without the need for manual cell sampling which risks introducing errors and heightening contamination risks.


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 field of biomimetic and bioinspired materials has progressed rapidly by drawing inspiration from nature's intricate structures and multifunctional systems in order to address pressing challenges in modern engineering. This review critically examines the mechanical performance of these materials, focusing on their hierarchical design principles, synthesis strategies, and versatility of application. Natural exemplars such as nacre, spider silk, and bone are emphasized, as their structural efficiency and functional adaptability have informed the development of synthetic analogues with superior strength, toughness, flexibility, and lightweight characteristics. The review also highlights advanced fabrication methods, such as additive manufacturing and precision chemical synthesis, which have enabled researchers to replicate the complexity of nature with ever‐greater accuracy. Key case studies demonstrate how bioinspired strategies have been translated into high‐performance materials for use in the aerospace, construction, and biomedical sectors. The review also discusses challenges related to scalability, reproducibility, and industrial integration. Finally, the review outlines emerging interdisciplinary approaches that are set to further enhance the mechanical properties and practical relevance of biomimetic materials, establishing them as transformative solutions for the next generation of engineering systems. Keywords: bioinspired design, biomimetic materials, engineering applications, mechanical performance, synthesis techniques

Candidates:
A. This review critically examines the mechanical performance of these materials, focusing on their hierarchical design principles, synthesis strategies, and versatility of application.
B. The review also highlights advanced fabrication methods, such as additive manufacturing and precision chemical synthesis, which have enabled researchers to replicate the complexity of nature with ever‐greater accuracy.
C. The evidence does not state that the review also highlights advanced fabrication methods, such as additive manufacturing and precision chemical synthesis, which have enabled researchers to replicate the complexity of nature with ever‐greater accuracy.
D. The evidence does not state that this review critically examines the mechanical performance of these materials, focusing on their hierarchical design principles, synthesis strategies, and versatility of application.
E. The field of biomimetic and bioinspired materials has progressed rapidly by drawing inspiration from nature's intricate structures and multifunctional systems in order to address pressing challenges in modern engineering.
F. The evidence does not state that the field of biomimetic and bioinspired materials has progressed rapidly by drawing inspiration from nature's intricate structures and multifunctional systems in order to address pressing challenges in modern engineering.
G. Natural exemplars such as nacre, spider silk, and bone are emphasized, as their structural efficiency and functional adaptability have informed the development of synthetic analogues with superior strength, toughness, flexibility, and lightweight characteristics.
H. Natural exemplars such as nacre, spider silk, and bone are not emphasized, as their structural efficiency and functional adaptability have informed the development of synthetic analogues with superior strength, toughness, flexibility, and lightweight characteristics.


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 develops and optimizes a high‐cell‐density cultivation (HCDC) of E. coli BL21 (DE3) to improve enzyme production utilizing DASGIP multibioreactor systems. By shifting from traditional shake flask methods to HCDC, we observed a 43‐fold increase in biomass production, with a comparable mass‐specific activity of the cell‐free extract. The HCDC demonstrated a 30‐fold increase in the active, soluble expression of arylmalonate decarboxylase (AMDase) from Bordetella bronchiseptica ( Bb AMDase) compared to conventional methodology. The protocol was designed to be readily adapted to various expression approaches and to provide a robust foundation for broader use in recombinant protein production. The HCDC was achieved with a linear glucose feed to promote robust cell growth. We optimized the glycerol feeding strategy during isopropyl β ‐D‐1‐thiogalactopyranoside (IPTG) induction to maximize AMDase production. Throughout the induction phase, we monitored both soluble expression yield and enzymatic activity, aiming to establish a process that is not only profitable but also efficient and stable. Keywords: arylmalonate decarboxylase, biocatalysis, E. coli BL21 (DE3), enzyme expression, high‐cell‐density cultivation

Candidates:
A. The HCDC demonstrated a 31‐fold increase in the active, soluble expression of arylmalonate decarboxylase (AMDase) from Bordetella bronchiseptica ( Bb AMDase) compared to conventional methodology.
B. By shifting from traditional shake flask methods to HCDC, we observed a 44‐fold increase in biomass production, with a comparable mass‐specific activity of the cell‐free extract.
C. The HCDC demonstrated a 30‐fold increase in the active, soluble expression of arylmalonate decarboxylase (AMDase) from Bordetella bronchiseptica ( Bb AMDase) compared to conventional methodology.
D. coli BL21 (DE3) to improve enzyme production utilizing DASGIP multibioreactor systems.
E. This study develops and optimizes a high‐cell‐density cultivation (HCDC) of E.
F. By shifting from traditional shake flask methods to HCDC, we observed a 43‐fold increase in biomass production, with a comparable mass‐specific activity of the cell‐free extract.
G. coli BL22 (DE3) to improve enzyme production utilizing DASGIP multibioreactor systems.
H. The evidence does not state that this study develops and optimizes a high‐cell‐density cultivation (HCDC) of E.


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:
Thermal oxo-degradation (TOD) of plastic can transform plastic waste into fermentable feedstocks. Bioconversion of the TOD products could be used as feedstocks in a biorefinery concept and lead to new avenues for plastic waste upcycling. Previous work demonstrated this concept with high-density polyethylene (HDPE), the most abundant type of plastic, and identified the nonconventional yeast Candida maltosa as a promising candidate for this application. Here, we describe the evolution of an improved strain of C. maltosa and characterize the uptake mechanisms of TOD products from HDPE (TOD_HDPE). Batch cultures in series passaged at the mid-exponential growth phase applied a selective pressure for faster growth and resulted in a >100% increase in specific growth rate when using TOD_HDPE as a carbon source compared to the wild-type strain used in previous work. Adaptive improvement in specific growth rate is an important step toward the development of an industrial strain, and it provides a basis for a mechanistic understanding of the TOD_HDPE uptake. The evolved strain was compared to the parent strain to identify the cellular and biochemical changes associated with the improved phenotype and the uptake mechanisms involved in the bioconversion of TOD_HDPE. This comparison found that C. maltosa secretes biosurfactants capable of solubilizing hydrocarbons. The adaptive evolution resulted in changes in biosurfactant production that translated to improved emulsification of alkanes and increased solubilization of fatty alcohols and alkanes. In addition to the changes in metabolites, the study identified increases in membrane permeability associated with a reduction in ergosterol that may also play a role in the improved phenotype. These findings support the development of C. maltosa and other potential microbial cell factories for plastic biorefineries and may inform future design strategies. Plastics are considered non-biodegradable because they take decades to break down into molecules that can enter the various carbon cycles. Thermal oxo-degradation can accelerate this rate-limiting step by turning plastic waste into a fermentable carbon source. In this work, we use Adaptive Laboratory Evolution to optimize the bioconversion of depolymerized HDPE by Candida maltosa . We also investigate the changes that resulted from the evolution process to learn about the molecular and cellular mechanisms involved in the bioconversion of hydrophobic substrates. Our findings show unique mechanisms in C. maltosa to overcome mass transfer limitations and metabolize long-chain hydrocarbons and fatty substrates that can possibly be extrapolated to industrial applications beyond plastic upcycling.

Candidates:
A. Previous work demonstrated this concept with high-density polyethylene (HDPE), the most abundant type of plastic, and identified the nonconventional yeast Candida maltosa as a promising candidate for this application.
B. The evidence does not state that previous work demonstrated this concept with high-density polyethylene (HDPE), the most abundant type of plastic, and identified the nonconventional yeast Candida maltosa as a promising candidate for this application.
C. Bioconversion of the TOD products could be used as feedstocks in a biorefinery concept and lead to new avenues for plastic waste upcycling.
D. Here, we describe the evolution of an improved strain of C.
E. The evidence does not state that here, we describe the evolution of an improved strain of C.
F. Thermal oxo-degradation (TOD) of plastic cannot transform plastic waste into fermentable feedstocks.
G. Thermal oxo-degradation (TOD) of plastic can transform plastic waste into fermentable feedstocks.
H. The evidence does not state that bioconversion of the TOD products could be used as feedstocks in a biorefinery concept and lead to new avenues for plastic waste upcycling.


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: ["61 g/L", "96 g/L", "81%", "60 g/L", "8 L", "80%", "95 g/L", "7 L"]

Evidence:
Bioplastic poly(3-hydroxybutyrate) (PHB) production by Halomonas bluephagenesis is typically activated under nitrogen limitation, inevitably restricting biomass formation and overall productivity. Here we identified gene ilvA encoding threonine deaminase as a flux-sensitive node linking branched-chain amino-acid synthesis to nitrogen sensing. Complete ilvA deletion or σ 54 disruption in H. bluephagenesis created a pseudo-nitrogen-limitation state that increased PHB accumulation yet concurrently suppressed microbial growth. To overcome this trade-off, two independent ilvA fine-tuning strategies were evaluated including synthetic sRNA-mediated translational repression and SspB/ClpXP proteolysis. In contrast, complete ilvA deletion created a pseudo-nitrogen-limited state that elevated PHB synthesis however reduced cell growth (cell dry weight or CDW) in 7 L bioreactors, reaching only 60 g/L CDW containing 80% PHB, far below the wild-type grown to 95 g/L CDW containing 60% PHB under same conditions, These results indicate that complete deletion of ilvA disrupts the balance between cell growth and PHB production. Under nitrogen rich fed-batch conditions, however, sRNA-based partial repression maintained CDW at 95 g/L while increasing PHB from 60% to 80 wt%, thereby establishing a growth-PHB production balance. These results suggest that controllable attenuation of ilvA , instead of full gene deletion, provides a potentially scalable approach for improving PHB production without severely compromising cell growth.


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:
2,4-Dihydroxybutyric acid (DHB) is a promising C 4 platform compound for the synthesis of methionine analogues and biodegradable polymers. However, aerobic DHB production from glucose in Escherichia coli involves transient acetate overflow prior to product synthesis, which could be challenging for process scalability. Therefore, we engineered Escherichia coli K-12 MG1655 for optimized DHB production by replacing the phosphotransferase system mediated glucose uptake with the galactose permease GalP, coupled to ATP-dependent phosphorylation via endogenous glucokinase. In combination with targeted deletions of malate- and fumarate-consuming reactions, we obtained a strain with enhanced flux through the tricarboxylic acid (TCA) cycle and pentose phosphate pathway leading to improved NADPH availability and increased anaplerotic activity, as revealed by 13 C metabolic flux analyses. Deletion of the mdh gene encoding for the cytosolic malate dehydrogenase further promoted DHB formation. The resulting strain achieved DHB yields up to 0.20 mol mol −1 (2.43 g L −1 ), a 4-fold increase compared to the wildtype background (0.05 mol mol −1 , 0.60 g L −1 ), under aerobic conditions while suppressing acetate formation. Together, these results demonstrate that GalP-mediated glucose uptake and engineering of the TCA cycle provide a robust metabolic framework for efficient DHB biosynthesis and establish a foundation for further process and pathway development. Keywords: 2,4-Dihydroxybutyric acid; Escherichia coli ; Metabolic engineering; Phosphotransferase system (PTS); Galactose permease (GalP); 13 C metabolic flux analysis

Candidates:
A. In combination with targeted deletions of malate- and fumarate-consuming reactions, we obtained a strain with enhanced flux through the tricarboxylic acid (TCA) cycle and pentose phosphate pathway leading to improved NADPH availability and increased anaplerotic activity, as revealed by 14 C metabolic flux analyses.
B. 3,4-Dihydroxybutyric acid (DHB) is a promising C 4 platform compound for the synthesis of methionine analogues and biodegradable polymers.
C. 2,4-Dihydroxybutyric acid (DHB) is a promising C 4 platform compound for the synthesis of methionine analogues and biodegradable polymers.
D. In combination with targeted deletions of malate- and fumarate-consuming reactions, we obtained a strain with enhanced flux through the tricarboxylic acid (TCA) cycle and pentose phosphate pathway leading to improved NADPH availability and increased anaplerotic activity, as revealed by 13 C metabolic flux analyses.
E. However, aerobic DHB production from glucose in Escherichia coli involves transient acetate overflow prior to product synthesis, which could be challenging for process scalability.
F. The evidence does not state that however, aerobic DHB production from glucose in Escherichia coli involves transient acetate overflow prior to product synthesis, which could be challenging for process scalability.
G. Therefore, we engineered Escherichia coli K-12 MG1655 for optimized DHB production by replacing the phosphotransferase system mediated glucose uptake with the galactose permease GalP, coupled to ATP-dependent phosphorylation via endogenous glucokinase.
H. Therefore, we engineered Escherichia coli K-13 MG1655 for optimized DHB production by replacing the phosphotransferase system mediated glucose uptake with the galactose permease GalP, coupled to ATP-dependent phosphorylation via endogenous glucokinase.


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:
Photobiocatalysis with photoautotrophic whole cells has demonstrated strong potential for producing chiral molecules and platform chemicals using sustainable inputs such as light, water and CO 2 under mild reaction conditions. Coupling enzymatic transformations directly to natural photosynthesis enables higher atom efficiency compared with heterotrophic systems. However, large‐scale application remains challenging, particularly due to light attenuation in photobioreactors. In this review, we summarize recent advances in whole‐cell photobiotransformations with emphasis on process conditions. We also discuss strategies for intensifying photobiocatalysis through improved reactor design and new immobilization materials, along with developments in fast‐growing photoautotrophic strains. Sustainability analyses indicate that organic electron donors represent only one factor influencing environmental performance, and simply replacing them with photosynthetic water splitting does not inherently yield a carbon‐negative process. Nonetheless, our calculations show that when high substrate loadings are combined with wastewater use and optimized downstream processing, photosynthesis‐driven biotechnology can offer substantial reductions in CO 2 emissions.

Candidates:
A. The evidence does not state that however, large‐scale application remains challenging, particularly due to light attenuation in photobioreactors.
B. The evidence does not state that in this review, we summarize recent advances in whole‐cell photobiotransformations with emphasis on process conditions.
C. Photobiocatalysis with photoautotrophic whole cells has demonstrated strong potential for producing chiral molecules and platform chemicals using sustainable inputs such as light, water and CO 3 under mild reaction conditions.
D. In this review, we summarize recent advances in whole‐cell photobiotransformations with emphasis on process conditions.
E. However, large‐scale application remains challenging, particularly due to light attenuation in photobioreactors.
F. Coupling enzymatic transformations directly to natural photosynthesis enables higher atom efficiency compared with heterotrophic systems.
G. Photobiocatalysis with photoautotrophic whole cells has demonstrated strong potential for producing chiral molecules and platform chemicals using sustainable inputs such as light, water and CO 2 under mild reaction conditions.
H. The evidence does not state that coupling enzymatic transformations directly to natural photosynthesis enables higher atom efficiency compared with heterotrophic systems.


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:
Naturally occurring in breast milk, human milk oligosaccharides (HMOs) are of great interest as an ingredient for infant nutrition due to numerous associated health benefits. Current commercial production relies mainly on microbial fermentation, while enzymatic synthesis is used to produce milligram scales for scientific studies. Enzymatic synthesis using glycosyltransferases and nucleotide sugars is especially promising due to high reaction yields, but is limited by low activity of glycosyltransferases and the high cost of nucleotide sugars. This study presents a novel approach that uses a single engineered E. coli BL21(DE3) strain to simultaneously express six recombinant enzymes (UMPK, PPK3, GALK, NAHK, GALU, and PPA). This enables dual‐nucleotide sugar synthesis through two integrated multienzyme cascades. The system uses cost‐effective substrates, including uridine 5′‐monophosphate (UMP), N ‐acetylglucosamine (GlcNAc), galactose (Gal), and ATP. In situ ATP regeneration is achieved through polyphosphate (PolyP n ) breakdown. Comparative studies of three different expression strain configurations demonstrated that crude cell lysate could serve as an effective biocatalyst. This eliminates the need for expensive enzyme purification while maintaining high catalytic activity. Using crude cell lysate, conversion yields approaching 100% were obtained. Both UDP‐GlcNAc and UDP‐Gal were successfully purified using anion‐exchange chromatography. Based on the UV spectrum, purities of 85–99% and recovery yields exceeding 90%, respectively, were achieved. The practical application of this system was demonstrated by the successful synthesis of two HMOs: Lacto‐ N ‐triose II (LNTII) and lacto‐ N ‐neotetraose (LNnT), which demonstrates an effective nucleotide sugar recycling in coupled enzymatic reactions and paves the way toward larger scale production.

Candidates:
A. Naturally occurring in breast milk, human milk oligosaccharides (HMOs) are not of great interest as an ingredient for infant nutrition due to numerous associated health benefits.
B. Current commercial production relies mainly on microbial fermentation, while enzymatic synthesis is used to produce milligram scales for scientific studies.
C. The evidence does not state that this study presents a novel approach that uses a single engineered E.
D. Naturally occurring in breast milk, human milk oligosaccharides (HMOs) are of great interest as an ingredient for infant nutrition due to numerous associated health benefits.
E. This study presents a novel approach that uses a single engineered E.
F. Enzymatic synthesis using glycosyltransferases and nucleotide sugars is not especially promising due to high reaction yields, but is limited by low activity of glycosyltransferases and the high cost of nucleotide sugars.
G. Enzymatic synthesis using glycosyltransferases and nucleotide sugars is especially promising due to high reaction yields, but is limited by low activity of glycosyltransferases and the high cost of nucleotide sugars.
H. Current commercial production relies mainly on microbial fermentation, while enzymatic synthesis is not used to produce milligram scales for scientific studies.


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 large‐scale use of toxic and environmentally hazardous solvents remains a major challenge in industrial manufacturing and consumer‐goods production. Conventional solubilization processes often depend on harsh conditions, including elevated temperatures and pressures, resulting in high energy consumption, health risks, and environmental pollution. Developing sustainable alternatives is therefore an urgent scientific and societal priority. This article discusses recent advances in green solubilization and emerging strategies aiming to reconcile efficiency with environmental compatibility. We address the future role of classical and “green” solvents, including ionic liquids (ILs) and natural deep eutectic solvents (NADES), and critically assess their benefits and limitations from a sustainability perspective. Particular emphasis is placed on water as the potentially “greenest” solvent, highlighting how its intrinsic tendency to form structured, heterogeneous environments can be advantageous or detrimental for solubilization. In this context, we examine mesoscale structuring, surfactant‐free microemulsions, and dynamic interfaces. Furthermore, naturally derived solubilizers such as hydrotropes, biosurfactants, and proteins are considered promising tools to enhance solubility while maintaining biocompatibility and low environmental impact. Selected examples from our own work illustrate how combining water‐based structuring with bio‐derived or benign additives can create new pathways toward energy‐efficient and sustainable solubilization technologies. Keywords: green solubilization, sustainable solubilization, sustainable chemistry, sustainable solvents

Candidates:
A. The evidence does not state that conventional solubilization processes often depend on harsh conditions, including elevated temperatures and pressures, resulting in high energy consumption, health risks, and environmental pollution.
B. Developing sustainable alternatives is therefore an urgent scientific and societal priority.
C. The evidence does not state that the large‐scale use of toxic and environmentally hazardous solvents remains a major challenge in industrial manufacturing and consumer‐goods production.
D. Developing sustainable alternatives is not therefore an urgent scientific and societal priority.
E. This article discusses recent advances in green solubilization and emerging strategies aiming to reconcile efficiency with environmental compatibility.
F. The evidence does not state that this article discusses recent advances in green solubilization and emerging strategies aiming to reconcile efficiency with environmental compatibility.
G. Conventional solubilization processes often depend on harsh conditions, including elevated temperatures and pressures, resulting in high energy consumption, health risks, and environmental pollution.
H. The large‐scale use of toxic and environmentally hazardous solvents remains a major challenge in industrial manufacturing and consumer‐goods production.


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

Evidence:
Processing of biomass residues for material and energy recovery generate effluents containing complex organic mixtures. Analyses of bulk parameters are conventional for characterization and classification of such effluents, where the limited information offered impedes the development of molecular level management practices. This study assessed the potential of solvent-selective complexity reduction of effluent dissolved organic matter (DOM) from anaerobic bioprocessing of biomass residues for 1 H NMR spectroscopy. The DOM were acquired after filtration and drying of samples from seven full scale anaerobic bioreactor facilities with effluents used as biofertilizer. The 1 H NMR spectra of DOM in indigenous solvent (water) revealed source dependent characteristics primarily due to variable abundance of aliphatic CCH in lipids and peptides, OCCH in carbohydrates, and olefinic and aromatic subunits. Dimethyl sulfoxide solubilized larger proportion of nonfunctionalized aliphatic and aromatic molecules, with 1 H NMR features also varying depending on the DOM source. Methanol, however, reduced the 1 H NMR spectral variability and dissolved sets of aliphatic and aromatic molecules from the dried DOM with similar 1 H NMR features irrespective of their origin. Among the other solvents studied, the reactive dissolution by trifluoroacetic acid decomposed aliphatic units while enriching aromatics (i.e., C ar H :CC H of 0.7 compared to 0.2 in water), also forming small (oligo)saccharides and peptide fragments. Acetone, dichloromethane, and acetonitrile extracted alkyl-rich molecules with varying degrees of functionalization. Acetonitrile separated a fraction enriched in aliphatic carboxylic acids, while dichloromethane mainly dissolved nonfunctionalized aliphatic hydrocarbons. It is proposed that a simple process of filtration, drying, and dissolution of effluent DOM in different solvents substantially reduces the complexity and heterogeneity of the organic mixtures enabling the structural discrimination of diverse molecular classes by 1 H NMR spectroscopy.

Candidates:
A. The DOM were not acquired after filtration and drying of samples from seven full scale anaerobic bioreactor facilities with effluents used as biofertilizer.
B. This study assessed the potential of solvent-selective complexity reduction of effluent dissolved organic matter (DOM) from anaerobic bioprocessing of biomass residues for 1 H NMR spectroscopy.
C. The DOM were acquired after filtration and drying of samples from seven full scale anaerobic bioreactor facilities with effluents used as biofertilizer.
D. The evidence does not state that processing of biomass residues for material and energy recovery generate effluents containing complex organic mixtures.
E. Analyses of bulk parameters are not conventional for characterization and classification of such effluents, where the limited information offered impedes the development of molecular level management practices.
F. This study assessed the potential of solvent-selective complexity reduction of effluent dissolved organic matter (DOM) from anaerobic bioprocessing of biomass residues for 2 H NMR spectroscopy.
G. Processing of biomass residues for material and energy recovery generate effluents containing complex organic mixtures.
H. Analyses of bulk parameters are conventional for characterization and classification of such effluents, where the limited information offered impedes the development of molecular level management practices.


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 in vitro engineering of vascularized cardiac tissues holds transformative potential for disease modeling, drug screening, and regenerative therapy. However, despite rapid advances in stem cell biology, biomaterials, and biofabrication technologies, the reconstruction of functional, perfusable vasculature within engineered myocardial tissues remains a central and unresolved challenge. In this review, we move beyond a descriptive catalog of available techniques and instead present a process‐oriented framework for understanding vascularized cardiac tissue engineering. By systematically analyzing how cellular components, biomaterial design, and biofabrication strategies collectively govern vascular formation, perfusion stability, and myocardial function, we examine self‐assembly, mold‐casting, 3D bioprinting, and microfluidic approaches, to critically evaluate their respective advantages and trade‐offs under cardiac‐specific physiological constraints. Finally, application prospects of vascularized cardiac tissues in disease modeling and drug testing are discussed, and current limitations and future directions are proposed to accelerate translational impact. By reframing vascularized cardiac tissue engineering as an integrated manufacturing challenge rather than a collection of isolated technologies, this review aims to provide a coherent conceptual guide for advancing functional human cardiac models. Keywords: cardiac cells, cardiac organoids, cardiac tissue engineering, regenerative medicine, vascularized cardiac tissues

Candidates:
A. The evidence does not state that in this review, we move beyond a descriptive catalog of available techniques and instead present a process‐oriented framework for understanding vascularized cardiac tissue engineering.
B. The in vitro engineering of vascularized cardiac tissues holds transformative potential for disease modeling, drug screening, and regenerative therapy.
C. By systematically analyzing how cellular components, biomaterial design, and biofabrication strategies collectively govern vascular formation, perfusion stability, and myocardial function, we examine self‐assembly, mold‐casting, 4D bioprinting, and microfluidic approaches, to critically evaluate their respective advantages and trade‐offs under cardiac‐specific physiological constraints.
D. However, despite rapid advances in stem cell biology, biomaterials, and biofabrication technologies, the reconstruction of functional, perfusable vasculature within engineered myocardial tissues remains a central and unresolved challenge.
E. The evidence does not state that the in vitro engineering of vascularized cardiac tissues holds transformative potential for disease modeling, drug screening, and regenerative therapy.
F. In this review, we move beyond a descriptive catalog of available techniques and instead present a process‐oriented framework for understanding vascularized cardiac tissue engineering.
G. By systematically analyzing how cellular components, biomaterial design, and biofabrication strategies collectively govern vascular formation, perfusion stability, and myocardial function, we examine self‐assembly, mold‐casting, 3D bioprinting, and microfluidic approaches, to critically evaluate their respective advantages and trade‐offs under cardiac‐specific physiological constraints.
H. The evidence does not state that however, despite rapid advances in stem cell biology, biomaterials, and biofabrication technologies, the reconstruction of functional, perfusable vasculature within engineered myocardial tissues remains a central and unresolved challenge.


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 composition of muscle fiber types and the development of skeletal muscle are critical determinants of cultured meat quality. L‐carnosine, a dipeptide abundant in ruminant muscle, is known to influence meat quality, yet its regulatory mechanisms in bovine skeletal muscle satellite cells (BSCs) for cultured meat production remain unclear. This study aimed to elucidate the effects of L‐carnosine on the proliferation, differentiation, and muscle fiber type transformation of Yanbian cattle BSCs. We identified 10 mM as the optimal concentration for enhancing cell proliferation ( p < 0.05), a key finding established by screening L‐carnosine treatments from 0 to 40 mm. This enhancement was mediated by the upregulation of cell cycle genes (Pax7, Ki67, CDK1, CDK2, PCNA) and the suppression of inhibitors ( p21, p53, p16 ). Furthermore, L‐carnosine robustly promoted myotube formation and specifically upregulated fast‐twitch muscle fiber markers (MyHC2a, MyHC2b, MyHC2x) while downregulating the slow‐twitch marker MyHC1 ( p < 0.05). Transcriptomic analysis identified 449 differentially expressed genes, which were significantly enriched in the PI3K‐Akt signaling pathway. Western blotting confirmed that L‐carnosine activates the Akt/mTOR/P70S6K signaling pathway to drive myogenesis. Additionally, L‐carnosine demonstrated significant antioxidant capacity by reducing reactive oxygen species (ROS) and lipid peroxidation (MDA) while enhancing antioxidant enzyme activities (SOD and GSH‐Px). In conclusion, this study provides the first evidence that L‐carnosine promotes BSC proliferation and fast‐twitch fiber differentiation via the Akt/mTOR/P70S6K pathway, suggesting its potential as a highly effective, natural additive for cultured meat production. Keywords: Akt/mTOR/P70S6K signaling pathway, antioxidant activity, L‐carnosine, proliferation and differentiation, skeletal muscle satellite cells, yanbian cattle

Candidates:
A. The composition of muscle fiber types and the development of skeletal muscle are not critical determinants of cultured meat quality.
B. L‐carnosine, a dipeptide abundant in ruminant muscle, is known to influence meat quality, yet its regulatory mechanisms in bovine skeletal muscle satellite cells (BSCs) for cultured meat production remain unclear.
C. We identified 11 mM as the optimal concentration for enhancing cell proliferation ( p < 0.05), a key finding established by screening L‐carnosine treatments from 0 to 40 mm.
D. We identified 10 mM as the optimal concentration for enhancing cell proliferation ( p < 0.05), a key finding established by screening L‐carnosine treatments from 0 to 40 mm.
E. The evidence does not state that this study aimed to elucidate the effects of L‐carnosine on the proliferation, differentiation, and muscle fiber type transformation of Yanbian cattle BSCs.
F. This study aimed to elucidate the effects of L‐carnosine on the proliferation, differentiation, and muscle fiber type transformation of Yanbian cattle BSCs.
G. The composition of muscle fiber types and the development of skeletal muscle are critical determinants of cultured meat quality.
H. L‐carnosine, a dipeptide abundant in ruminant muscle, is not known to influence meat quality, yet its regulatory mechanisms in bovine skeletal muscle satellite cells (BSCs) for cultured meat production remain unclear.


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:
Efficient and scalable production of viral antigens remains a key challenge in the development of recombinant subunit vaccines and diagnostic reagents. Microbial expression systems, particularly Pichia pastoris ( P. pastoris ), offer a promising platform for producing complex viral glycoproteins with appropriate folding and post-translational modifications. In this study, the hemagglutinin head domain (HA1) of influenza A (H1N1) virus was expressed as a secreted recombinant protein in P. pastoris GS115. The HA1 gene was codon-optimized and expressed under the control of the methanol-inducible AOX1 promoter with an α-factor signal peptide. Multicopy integrants were enriched using G418 selection, and expression conditions were systematically optimized. Under shake-flask induction, the selected recombinant strain produced up to 0.375 g/L of rHA1 in the culture supernatant. The protein was efficiently purified by Ni-NTA affinity chromatography to a purity exceeding 95%. PNGase F digestion confirmed N-linked glycosylation. Limited functional validation demonstrated that the yeast-expressed rHA1 retained antigenic integrity, as evidenced by the induction of rHA1-specific antibodies and hemagglutination-inhibiting activity in a murine model. These results establish P. pastoris as an effective microbial cell factory for the high-level secretion of influenza HA1 protein. The optimized expression and purification strategy provides a scalable and cost-efficient framework for microbial production of viral antigens and may be applicable to other glycoproteins of biomedical relevance. Keywords: Pichia pastoris , recombinant protein expression, hemagglutinin HA1, secretory expression, microbial cell factory, viral glycoprotein


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 quantification of lentiviral vector (LVV) potency via titration is a critical quality control step for cell and gene therapies. However, standard functional titre assays are fundamentally limited by an inability to detect multiple integration events, procedural variations, and, most critically, a mass transport limitation created by the fluid overlay in conventional well plates. These issues, compounded by a lack of standardisation, lead to significant inter-laboratory variability and a systematic underestimation of true vector potency. In this study, we employed a microfluidic approach to create a more precisely engineered assay environment. We systematically evaluated channel depth, incubation time, vector concentration, and multiplicity of infection (MOI) for their impact on assay linearity, sensitivity, limit of detection, and reproducibility. A 0.2 mm deep channel provided linearity and reproducibility comparable to 96-well plates yet offered shorter incubation periods, and enhanced sensitivity, detecting activity down to a MOI of 0.0625 (corroborated by qPCR analysis) - a level at which conventional well plates fail. This establishes our microfluidic platform as a device-based analytical standard that transforms functional titre quantification from a variable protocol into a more reliable engineering solution for quality testing in cell therapy manufacturing.

Candidates:
A. However, standard functional titre assays are not fundamentally limited by an inability to detect multiple integration events, procedural variations, and, most critically, a mass transport limitation created by the fluid overlay in conventional well plates.
B. These issues, compounded by a lack of standardisation, lead to significant inter-laboratory variability and a systematic underestimation of true vector potency.
C. However, standard functional titre assays are fundamentally limited by an inability to detect multiple integration events, procedural variations, and, most critically, a mass transport limitation created by the fluid overlay in conventional well plates.
D. The quantification of lentiviral vector (LVV) potency via titration is a critical quality control step for cell and gene therapies.
E. The evidence does not state that these issues, compounded by a lack of standardisation, lead to significant inter-laboratory variability and a systematic underestimation of true vector potency.
F. In this study, we employed a microfluidic approach to create a more precisely engineered assay environment.
G. The quantification of lentiviral vector (LVV) potency via titration is not a critical quality control step for cell and gene therapies.
H. The evidence does not state that in this study, we employed a microfluidic approach to create a more precisely engineered assay environment.


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 basement membrane is a specialized extracellular matrix that compartmentalizes epithelial and endothelial tissues and provides essential structural and signaling cues for tissue organization. Whereas fibrillar collagens (Col) of the interstitial matrix (such as Col I and Col III) are widely used in tissue modeling, the networking collagens that scaffold the basement membrane, including human Col IV and Col VI, remain difficult to access. Commercial basement membrane surrogates such as Matrigel® are derived from murine tumors and are ill defined, dilute, variable, and incompatible with animal-free biomanufacturing. Thus, there is a crucial need for human-derived basement membrane matrices that are free of xenogenic contaminants and do not rely on breeding animals. Here, we analyzed whether human mesenchymal stromal cells (MSCs) could serve as a platform to produce self-assembling basement membrane components under chemically defined, xeno-free conditions. MSCs from placental, umbilical cord, bone marrow, and adipose tissues were cultured as three-dimensional spheroids and adherent multilayered sheets. Confocal imaging of whole-mount, decellularized spheroid matrices showed complex networks of fibronectin (FN) and Col IV with topological and organizational features characteristic of basement membrane. Perinatal MSCs produced distinct matrix architectures consisting of apical FN sheets underlaid by continuous Col IV networks. Time-resolved imaging of umbilical cord MSC-derived matrix sheets demonstrated a reproducible sequence of basement membrane assembly that parallels developmental tissue organization. Together, these findings demonstrate that human MSCs cultured entirely without entirely animal-derived components can synthesize functional basement membrane proteins that self-assemble into ordered, tissue-like scaffolds. In this work, we establish MSCs as a scalable, sustainable, and cruelty-free platform for manufacturing human basement membrane matrices for bioengineering and regenerative medicine applications.

Candidates:
A. The basement membrane is not a specialized extracellular matrix that compartmentalizes epithelial and endothelial tissues and provides essential structural and signaling cues for tissue organization.
B. The basement membrane is a specialized extracellular matrix that compartmentalizes epithelial and endothelial tissues and provides essential structural and signaling cues for tissue organization.
C. Commercial basement membrane surrogates such as Matrigel® are not derived from murine tumors and are ill defined, dilute, variable, and incompatible with animal-free biomanufacturing.
D. Whereas fibrillar collagens (Col) of the interstitial matrix (such as Col I and Col III) are not widely used in tissue modeling, the networking collagens that scaffold the basement membrane, including human Col IV and Col VI, remain difficult to access.
E. Whereas fibrillar collagens (Col) of the interstitial matrix (such as Col I and Col III) are widely used in tissue modeling, the networking collagens that scaffold the basement membrane, including human Col IV and Col VI, remain difficult to access.
F. Commercial basement membrane surrogates such as Matrigel® are derived from murine tumors and are ill defined, dilute, variable, and incompatible with animal-free biomanufacturing.
G. Thus, there is not a crucial need for human-derived basement membrane matrices that are free of xenogenic contaminants and do not rely on breeding animals.
H. Thus, there is a crucial need for human-derived basement membrane matrices that are free of xenogenic contaminants and do not rely on breeding animals.


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:
Laccases are multicopper enzymes capable of oxidizing a wide variety of compounds, standing out as green tools for industrial and environmental applications. However, production from native sources faces limitations that have driven advances in recombinant expression. This scoping review includes studies published between 2019 and 2025, selected from major online databases, describing recombinant fungal and bacterial laccases for environmental applications. Strategies to optimize expression are discussed, including the use of efficient vectors, codon optimization, His-tag addition, mutagenesis, and computational approaches, with an emphasis on their advantages, trade-offs, and limitations. Pichia pastoris is widely used for the expression of fungal laccases, while Escherichia coli is preferred for the expression of bacterial laccases. However, significant variability in expression efficiency and enzyme performance is observed across hosts and constructs. The use of alternative culture media, such as agro-industrial residues, is also explored as a sustainability-driven strategy; however, its applicability remains limited for certain heterologous expression systems. In the environmental field, recombinant laccases demonstrate high efficiency in the degradation of textile dyes, the treatment of lignocellulosic waste, the biodegradation of pharmaceuticals, and the degradation of various toxic compounds, often requiring redox mediators to achieve high conversion rates. Despite significant advances, challenges remain, such as inconsistent catalytic performance among studies and limited stability under extreme temperature and pH conditions. Overall, this review highlights the key challenges in developing recombinant laccase and demonstrates that advances in protein engineering, expression systems, and process optimization are crucial for environmental applications.

Candidates:
A. Laccases are not multicopper enzymes capable of oxidizing a wide variety of compounds, standing out as green tools for industrial and environmental applications.
B. This scoping review includes studies published between 2020 and 2025, selected from major online databases, describing recombinant fungal and bacterial laccases for environmental applications.
C. Strategies to optimize expression are not discussed, including the use of efficient vectors, codon optimization, His-tag addition, mutagenesis, and computational approaches, with an emphasis on their advantages, trade-offs, and limitations.
D. This scoping review includes studies published between 2019 and 2025, selected from major online databases, describing recombinant fungal and bacterial laccases for environmental applications.
E. The evidence does not state that however, production from native sources faces limitations that have driven advances in recombinant expression.
F. However, production from native sources faces limitations that have driven advances in recombinant expression.
G. Laccases are multicopper enzymes capable of oxidizing a wide variety of compounds, standing out as green tools for industrial and environmental applications.
H. Strategies to optimize expression are discussed, including the use of efficient vectors, codon optimization, His-tag addition, mutagenesis, and computational approaches, with an emphasis on their advantages, trade-offs, and limitations.


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: ["+54%", "+66%", "+53%", "+65%"]

Evidence:
In large‐scale bioprocesses, mixing limitations and design constraints cause the onset of heterogeneous environments, subjecting the cells to continuously changing external conditions, often reducing their performance compared to laboratory conditions. This study evaluated the performance in producing a heterologous transaminase (TA) of a genome‐reduced Escherichia coli strain (RM214) in a STR‐PFR scale‐down system, benchmarking it against a wild‐type strain. Under cycles of glycerol limitation and starvation, combined with oxygen limitation in later process stages, RM214 outperformed the wild‐type strain. Due to its lower maintenance coefficient, RM214 showed a remarkable biomass increase of +53% and a boosted final volumetric activity with a +65% increase. These results were achieved with significantly reduced biomass‐specific substrate uptake rates and respiratory parameters, both crucial for optimizing large‐scale processes. This study underscores the applicability and enhanced robustness of genome‐reduced strains in heterogeneous large‐scale environments. Keywords: bioprocess scale‐up, bioreactor gradients, genome‐reduced strain, heterologous enzyme production, scale‐down system, transaminase


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:
Fish waste disposal poses significant environmental and economic challenges, limiting sustainability in the marine food industry. Hence, sustainable valorization strategies are needed to enhance resource recovery while minimizing waste. As an approach, this study aimed to evaluate the potential of converting chum salmon ( Oncorhynchus keta ) head (CSH) waste into a high-value peptone for microbiological applications. Various proteolytic enzymes were screened for CSH hydrolysis, among which Protamex achieved the highest hydrolysis and recovery rates. The resulting chum salmon head peptone (CSHP) exhibited favorable characteristics, including a low average molecular mass (557 Da) and a high amino nitrogen content (4.9%), outperforming commercial animal (AP) and vegetable (VP) peptones. To assess its biotechnological potential, CSHP was evaluated as a nitrogen source for recombinant protein production and supported higher expression of human superoxide dismutase (hSOD) and human growth hormone (hGH) in Escherichia coli BL21(DE3), compared with AP, VP, and Luria–Bertani (LB) media. Furthermore, life cycle assessment revealed a substantially lower carbon footprint for CSHP production than that of conventional peptone sources. These findings suggest that CSHP is a reliable and sustainable alternative to traditional peptones, offering both therapeutic and industrial applications while contributing to marine waste reduction and circular bioeconomy strategies.

Candidates:
A. Various proteolytic enzymes were not screened for CSH hydrolysis, among which Protamex achieved the highest hydrolysis and recovery rates.
B. The evidence does not state that as an approach, this study aimed to evaluate the potential of converting chum salmon ( Oncorhynchus keta ) head (CSH) waste into a high-value peptone for microbiological applications.
C. Various proteolytic enzymes were screened for CSH hydrolysis, among which Protamex achieved the highest hydrolysis and recovery rates.
D. Hence, sustainable valorization strategies are not needed to enhance resource recovery while minimizing waste.
E. The evidence does not state that fish waste disposal poses significant environmental and economic challenges, limiting sustainability in the marine food industry.
F. Fish waste disposal poses significant environmental and economic challenges, limiting sustainability in the marine food industry.
G. Hence, sustainable valorization strategies are needed to enhance resource recovery while minimizing waste.
H. As an approach, this study aimed to evaluate the potential of converting chum salmon ( Oncorhynchus keta ) head (CSH) waste into a high-value peptone for microbiological applications.


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

Evidence:
The growing global population and increasing pressure on conventional food systems have intensified the search for sustainable and nutrient-rich protein sources. Blue foods derived from marine and freshwater organisms offer significant nutritional advantages and lower environmental footprints compared with many terrestrial animal proteins. However, challenges related to resource sustainability, processing, preservation, and product traceability limit their full potential. This review provides a broad overview of emerging technologies shaping the future of blue food systems, covering innovative production strategies, advanced processing techniques, and omics-based analytical approaches. Key developments in cellular aquaculture and cellular mariculture are discussed as promising alternatives to traditional fisheries and aquaculture, enabling the production of blue food through controlled cell cultivation. Additionally, alternative protein platforms including plant-based, fermentation-derived, and cultivated blue food analogues are assessed for their potential to enhance sustainability and diversify aquatic protein sources. Advanced structuring technologies such as extrusion, electrospinning, wet spinning, and 3D printing are highlighted for their roles in developing blue food analogues with improved texture and sensory attributes. Furthermore, non-thermal preservation techniques, including cold plasma (CP), high-pressure processing (HPP), pulsed electric fields (PEFs), and ultraviolet-based treatments, are reviewed for their effectiveness in improving microbial safety and extending shelf life while maintaining nutritional quality. The integration of omics technologies (proteomics, metabolomics, and lipidomics) provides deeper molecular insights into product quality, authenticity, and traceability within blue food supply chains. Collectively, these interdisciplinary advancements demonstrate strong potential to transform blue food production into a more resilient, sustainable, and technology-driven sector. Future progress will depend on overcoming challenges related to scalability, regulatory frameworks, and consumer acceptance to enable the successful commercialization of next-generation blue food products.

Candidates:
A. The evidence does not state that this review provides a broad overview of emerging technologies shaping the future of blue food systems, covering innovative production strategies, advanced processing techniques, and omics-based analytical approaches.
B. Blue foods derived from marine and freshwater organisms offer significant nutritional advantages and lower environmental footprints compared with many terrestrial animal proteins.
C. The evidence does not state that however, challenges related to resource sustainability, processing, preservation, and product traceability limit their full potential.
D. This review provides a broad overview of emerging technologies shaping the future of blue food systems, covering innovative production strategies, advanced processing techniques, and omics-based analytical approaches.
E. However, challenges related to resource sustainability, processing, preservation, and product traceability limit their full potential.
F. The growing global population and increasing pressure on conventional food systems have intensified the search for sustainable and nutrient-rich protein sources.
G. The evidence does not state that blue foods derived from marine and freshwater organisms offer significant nutritional advantages and lower environmental footprints compared with many terrestrial animal proteins.
H. The evidence does not state that the growing global population and increasing pressure on conventional food systems have intensified the search for sustainable and nutrient-rich protein sources.