Pilot scale manufacturing of black seed oil-loaded alginate beads; process development, and stability of thymoquinone.

The approach of ionic gelation was employed at the pilot scale of the 50 kg batch size to manufacture black seed oil (BSO)-loaded alginate (ALG) beads as a natural source supplementing the main bioactive compound of BSO, i.e., thymoquinone (TQ). The BSO-ALG emulsion was prepared by initially emulsifying BSO with alginate solution at the pilot scale in two stages. The final emulsion was then dripped through 12 units of 3D-printed multi-nozzles into a curing bath containing Ca2+. The dripping flow rate was scaled up to 288 mL/min through the 3D-printed multi-nozzles (22-gauge). The characteristics of pilot scale BSO-ALG beads were similar to those produced at the lab scale; the beads were spherical with a size of 1.84-1.94 mm. The mechanical strength and loss on drying ranged from 143.6 to 172 g and 77.85-81.96 %, respectively. The production yield and encapsulation efficiency were 77.53-83.65 % and 95.36-97.9 %, respectively. Furthermore, the emulsification process did not affect TQ stability, while the curing process reduced TQ concentration from 1.51 % to 1.03 % w/w. Additionally, a substantial drop in TQ concentration in the encapsulated BSO was observed after the drying process, where it reached 0.23 % w/w. Finally, the stability of BSO-ALG beads in both wet and dried forms under real-time and accelerated conditions for 3 months revealed that beads were stable in terms of their organoleptic characteristics, size and sphericity, and loss on drying. Findings from this study enable the large-scale manufacturing of encapsulated BSO and similar bioactive compounds in ALG beads for the first time. These findings are valuable for advancing microencapsulation through ionic gelation and enhancing food preservation and safety.

From agricultural biomass to D form lactic acid in ton scale via strain engineering of Lactiplantibacillus pentosus.

Worsening environmental conditions make lactic acid a sustainable alternative to petroleum-based plastics. This study created a genetically-engineered strain Lactiplantibacillus pentosus PeL containing a disrupted L-lactate dehydrogenase gene to produce high yield and optically pure D-lactic acid. Cellobiose was identified as the optimal sugar in the single carbon source test, yielding the highest lactic acid. In 5-L fermentation tests, pretreated wood chips hydrolysate was the best lignocellulosic substrate for PeL, resulting in a D-lactic acid yield of 900.7 ±â€¯141.4 mg/g of consumed sugars with an optical purity of 99.8 ±â€¯0.0 %. Gradually scaled-up fermentations using this substrate were achieved in 100-, and 9,000-L fermenters; PeL produced remarkably high D-lactic acid yields of 836.3 ±â€¯11.9 and 915.9 ±â€¯4.4 mg/g of consumed sugars, with optical purities of 95.0 ±â€¯0.0 % and 93.8 ±â€¯0.2 %, respectively. This study is the pioneer in demonstrating economical and sustainable ton-scale production of D-lactic acid.

Scalable Multistep Imprinting of Multiplexed Optical Anti-counterfeiting Patterns with Hierarchical Structures.

Multiplexed optical techniques with multichannel patterns provide powerful strategies for high-capacity anti-counterfeiting. However, it is still a big challenge to meet the demands of achieving high encryption levels, excellent readability, and simple preparation simultaneously. Herein, we use a multistep imprinting technique, leveraging surface work-hardening to massively produce multiplexed encrypted patterns with hierarchical structures. These patterns with coupled nano- and microstructures can be instantaneously decoded into different pieces of information at different view angles under white light illumination. By incorporating perpendicular nano- and microgratings, we achieve four-channel encoded patterns, enhancing anti-counterfeiting capacity. This versatile method works on various metal/polymer materials, offering high-density information storage, direct visibility, broad material compatibility, and low-cost mass production. Our high-performance anti-counterfeiting patterns show significant potential in real-world applications.

Pretreated sugarcane bagasse matches performance of synthetic media for lipid production with Yarrowia lipolytica.

Engineered strains of Yarrowia lipolytica with modified lipid profiles and other desirable properties for microbial oil production are widely reported but are almost exclusively characterized in synthetic laboratory-grade media. Ensuring translatable performance between synthetic media and industrially scalable lignocellulosic feedstocks is a critical challenge. Yarrowia lipolytica growth and lipid production were characterized in media derived from two-step acid-catalyzed glycerol pretreatment of sugarcane bagasse. Fermentation performance was benchmarked against laboratory-grade synthetic growth media, including detailed characterization of media composition, nitrogen utilization, biomass and lipid production, and fatty acid product profile. A Yarrowia lipolytica strain modified to enable xylose consumption consumed all sugars, glycerol, and acetic acid, accumulating lipids to 34-44 % of cell dry weight. Growth and lipid content when grown in sugarcane bagasse-derived media were equivalent to or better than that observed with synthetic media. These sugarcane bagasse-derived media are suitable for transferable development of Yarrowia lipolytica fermentations from synthetic media.

Acoustic resonance technology and quality by design approach facilitate the development of the robust tetrandrine nano-delivery system.

The aim of this study was to develop a sufficiently robust tetrandrine (Tet) nano-delivery system using acoustic resonance (AR) technology and freeze-drying technology. This system can effectively improve the solubility and dissolution properties of Tet, along with high stability and scale-up adaptability. Firstly, 54 stabilizers were screened simultaneously in a high-throughput manner with the help of AR technology to fully explore the optimal prescription space of tetrandrine nanosuspension (Tet-NS). The Plackett-Burman design was used to screen for critical variables severely affecting the quality of Tet-NS. The Box-Behnken design was used to investigate and optimize critical variables to obtain optimal nanosuspensions. The optimal prescription was successfully scaled up by 100 times, which was the initial exploration of its commercial scale production. Solidification studies have shown that formulations with 2.44% fructose as the cryoprotectant have excellent redispersibility. Compared with pure Tet, Tet in Tet-NS showed a significant increase in solubility and dissolution rate in water. Fourier transform infrared (FT-IR) demonstrated that no significant interactions occurred between the drug and excipients in Tet-NS. Powder x-ray diffraction analysis (PXRD) indicated that some of the Tet transformed into amorphous state during the preparation process. In short-term stability study, Tet-NS successfully maintained its physical stability. In summary, under the guidance of the QbD concept, this study rapidly developed Tet-NS using acoustic resonance technology, which can effectively improve the solubility and dissolution properties of Tet. During the development of Tet-NS, AR technology has demonstrated high particle size reduction capability, the ability to process multiple sets of formulations in parallel, and excellent scale-up capability. Meanwhile, the method and concept of this study are not limited to Tet, but also applicable to other poorly water-soluble drugs.

A framework for early-stage sustainability assessment of innovation projects enabled by weighted sum multi-criteria decision analysis in the presence of uncertainty.

A two-level hierarchical framework for early-stage sustainability assessment (FESSA) amongst a set of alternatives applicable from the earliest stages of process or product development is introduced, and its use in combination with an improved method weighted-sum method multi-criteria decision analysis (WSM-MCDA) in the presence of uncertainty is presented through application to a case study based upon a real-world decision scenario from speciality polymer manufacture. The approach taken addresses the challenge faced by those responsible for innovation management in the manufacturing process industries to make simultaneously timely and rational decisions early in the innovation cycle when knowledge gaps and uncertainty about the options tend to be at their highest. The Computed Uncertainty Range Evaluations (CURE) WSM-MCDA method provides better discrimination than the existing Multiple Attribute Range Evaluations (MARE) method without the computational burden of generating heuristic outcome distributions via Monte-Carlo simulation.

This paper introduces a framework that teams can use to think systematically about the wide range of criteria which go into deciding whether a proposed innovation enhances sustainability or not and shows how an improved method for multiple-criteria decision analysis can be used to put it into practice with an example drawn from the speciality chemicals industry. Innovation in the manufacturing process industries requires decisions to be made. In individual projects, scientists and technical managers must decide which technology, materials, and equipment to use. Equally, those responsible for directing a portfolio of projects must choose which projects to prioritise. In either case, early decision making is desirable to avoid sinking time and money into dead-end projects, and to identify what further work is needed for projects with a future. The earlier you decide however, the harder it can be to obtain firm evidence (e.g. conclusive experimental data, fully validated costings, or life cycle impacts) upon which to base your decision. The growing societal expectation that sustainability criteria are factored into such decisions merely adds to the challenges faced by the decision maker. Decisions must be made upon the evidence that is available combined with the informed judgement of those with knowledge of the system under consideration. This is best approached as a facilitated, team-based activity where assertions, assumptions and interpolations or extrapolations from the limited data can be tested and challenged. A sound decision-making process needs a suitable computational method for turning this complex qualitative and semi-quantitative assessment into a clear output indicator of potential success or failure for the options under consideration. The method described in this paper addresses this need but, just as importantly, the methodology ensures that the thought process behind whatever decision is indicated is clearly and transparently documented for future reference.

NaOH-Assisted Multicomponent Reaction and Polymerizations of Elemental Sulfur, Diisocyanides, and Diols to Access Functional Poly(O-thiocarbamate)s.

Sulfur-containing polymers with unique structures and fascinating properties have attracted much attention recently, the efficient and economic synthetic approaches for various sulfur-containing polymers have rapidly developed. Herein, the multicomponent reaction of elemental sulfur, isocyanide, and alcohol was designed at mild condition with the assistance of NaOH, and the corresponding NaOH-assisted multicomponent polymerization of elemental sulfur, diisocyanides, and diols were developed at room temperature or 40 oC in air, to produce poly(O-thiocarbamate)s with well-defined structures, high molecular weights (Mws up to 32 500 g/mol) and high yields (up to 99%). The facilely available monomers, mild condition, and high efficiency of this MCP enabled scale-up synthesis of poly(O-thiocarbamate)s, and 7.34 g polymer was obtained in 98% yield. These functional poly(O-thiocarbamate)s could enrich Au3+ from aqueous solution with high enrichment capacity (983 mg·Au3+/g) and high efficiency (>99.99%) in 1 min, demonstrating superior gold enrichment performance and their potential industrial and economic values.

Catalyst Design and Engineering for CO2-to-Formic Acid Electrosynthesis for a Low-Carbon Economy.

Formic acid (FA) has emerged as a promising candidate for hydrogen energy storage due to its favorable properties such as low toxicity, low flammability, and high volumetric hydrogen storage capacity under ambient conditions. Recent analyses have suggested that FA produced by electrochemical carbon dioxide (CO2) reduction reaction (eCO2RR) using low-carbon electricity exhibits lower fugitive hydrogen (H2) emissions and global warming potential (GWP) during the H2 carrier production, storage and transportation processes compared to those of other alternatives like methanol, methylcyclohexane, and ammonia. eCO2RR to FA can enable industrially relevant current densities without the need for high pressures, high temperatures, or auxiliary hydrogen sources. However, the widespread implementation of eCO2RR to FA is hindered by the requirement for highly stable and selective catalysts. Herein, the aim is to explore and evaluate the potential of catalyst engineering in designing stable and selective nanostructured catalysts that can facilitate economically viable production of FA.

Scaling up healthy eating in early childhood education and care: evaluation of the Appetite to Play capacity-building intervention.

OBJECTIVE: The purpose of this study was to examine the dissemination of the healthy eating component of Appetite to Play at scale using the Reach, Effectiveness, Adoption, Implementation, Maintenance (RE-AIM) framework. DESIGN: The Appetite to Play capacity-building intervention is a set of evidence-informed implementation strategies aimed at enhancing the adoption of recommended practices for promoting healthy eating and active play in early years settings. The evaluation was pragmatic, employing both quantitative (surveys) and qualitative (interviews) data collection. SETTING: The Appetite to Play intervention was delivered through in-person community-based workshops, virtual workshops, asynchronous e-learning and online resources. PARTICIPANTS: We received completed surveys from 1670 in-person workshop participants (96 % female), and twenty-three (all female) survey respondents also participated in a telephone interview. Approximately two-thirds of all participant groups were certified early childhood educators. RESULTS: Results indicated that Appetite to Play had high reach (25 867 individual website visits, 195 workshops delivered), effectiveness (significant increases in care provider's knowledge, confidence (P < 0·05) and high post-intervention intention to implement), adoption (11 % of educators in BC trained) and implementation (good alignment with implementation strategies and current practices), with a significant maintenance plan to support the intervention's future success. CONCLUSIONS: An evidence-based capacity-building intervention with an emphasis on training and provision of practical online resources can improve early years providers' knowledge, confidence and intention to implement recommended practices that promote healthy eating. Further research is needed to determine the impact on child-level outcomes and how parents can be supported in contributing to positive food environments.

Challenges and opportunities for large-scale applications of the electro-Fenton process.

As an electrochemical advanced oxidation process, the electro-Fenton (EF) process has gained significant importance in the treatment of wastewater and persistent organic pollutants in recent years. As recently reported in a bibliometric analysis, the number of scientific publications on EF have increased exponentially since 2002, reaching nearly 500 articles published in 2022 (Deng et al., 2022). The influence of the main operating parameters has been thoroughly investigated for optimization purposes, such as type of electrode materials, reactor design, current density, and type and concentration of catalyst. Even though most of the studies have been conducted at a laboratory scale, focusing on fundamental aspects and their applications to degrade specific pollutants and treat real wastewater, important large-scale attempts have also been made. This review presents and discusses the most recent advances of the EF process with special emphasis on the aspects more closely related to future implementations at the large scale, such as applications to treat real effluents (industrial and municipal wastewaters) and soil remediation, development of large-scale reactors, costs and effectiveness evaluation, and life cycle assessment. Opportunities and perspectives related to the heterogeneous EF process for real applications are also discussed. This review article aims to be a critical and exhaustive overview of the most recent developments for large-scale applications, which seeks to arouse the interest of a large scientific community and boost the development of EF systems in real environments.

Innovations in one-step point-of-care testing within microfluidics and lateral flow assays for shaping the future of healthcare.

Point-of-care testing (POCT) technology, using lateral flow assays and microfluidic systems, facilitates cost-effective diagnosis, timely treatment, ongoing monitoring, and prevention of life-threatening outcomes. Aside from significant advancements demonstrated in academic research, implementation in real-world applications remains frustratingly limited. The divergence between academic developments and practical utility is often due to factors such as operational complexity, low sensitivity and the need for trained personnel. Taking this into consideration, our objective is to present a critical and objective overview of the latest advancements in fully integrated one-step POCT assays for home-testing which would be commercially viable. In particular, aspects of signal amplification, assay design modification, and sample preparation are critically evaluated and their features and medical applications along with future perspective and challenges with respect to minimal user intervention are summarized. Associated with and very important for the one-step POCT realization are also readout devices and fabrication processes. Critical analysis of available and useful technologies are presented in the SI section.

Unraveling the complexity of amorphous solid as direct ingredient for conventional oral solid dosage form: The story of Elagolix Sodium.

Conventional solid oral dosage form development is not typically challenged by reliance on an amorphous drug substance as a direct ingredient in the drug product, as this may result in product development hurdles arising from process design and scale-up, control of physical quality attributes, drug product processability and stability. Here, we present the Chemistry, Manufacturing and Controls development journey behind the successful commercialization of an amorphous drug substance, Elagolix Sodium, a first-in-class, orally active gonadotropin-releasing hormone antagonist. The reason behind the lack of crystalline state was assessed via Molecular Dynamics (MD) at the molecular and inter-molecular level, revealing barriers for nucleation due to prevalence of intra-molecular hydrogen bond, repulsive interactions between active pharmaceutical ingredient (API) molecules and strong solvation effects. To provide a foundational basis for the design of the API manufacturing process, we modeled the solvent-induced plasticization behavior experimentally and computationally via MD for insights into molecular mobility. In addition, we applied material science tetrahedron concepts to link API porosity to drug product tablet compressibility. Finally, we designed the API isolation process, incorporating computational fluid dynamics modeling in the design of an impinging jet mixer for precipitation and solvent-dependent glass transition relationships in the cake wash, blow-down and drying process, to enable the consistent manufacture of a porous, non-sintered amorphous API powder that is suitable for robust drug product manufacturing.

A new strategy in bioreactor scale-up and process transfer using a dynamic initial vvm according to different aeration pore size.

Monoclonal antibody drugs have grown into a drug category with a market size of over $100 billion since the first product was launched on the market, which naturally creates a large demand for production. At the same time, the $100 billion market is distributed among more than 200 listed drugs, which indicates that the production demand for monoclonal antibody drugs is diverse. To meet this demand, major suppliers offer single-use bioreactors of all sizes. These single-use bioreactors with different specifications, especially the inconsistency of aeration pore sizes, pose great challenges for technology transfer and scale-up production, and the conventional scale-up strategies of constant Power input/volume ratio (P/V) and constant vessel volume per minute (vvm) can no longer meet the needs. This study simplified the selection of technical parameters in bioreactors based on the differences in aeration pore size. Innovatively combined the aeration pore sizes with initial aeration vvm, and comprehensively investigated the relationship between P/V, vvm and aeration pore size by designing experiments (DoE) using the orthogonal test method. The results showed a quantitative relationship between the aeration pore size and the initial aeration vvm in the P/V range of 20 ± 5 W/m3. The appropriate initial aeration was between 0.01 and 0.005 m3/min for aeration pore size ranging from 1 to 0.3 mm, which was the optimal incubation condition in the bioreactors. The choice of initial ventilation was most related to the final expression. Follow-up studies validated these findings in a 15 L glass bioreactor and a 500 L single-use bioreactor, and the results were consistent with expectations.

Real-world public health interventions demonstrate how research evidence informs program scale-up.

Evidence-based population interventions rely on intervention testing (efficacy and effectiveness trials) to determine what works to improve public health. We investigated the characteristics of real-world public health interventions to address obesity and explored the extent to which research testing was undertaken prior to scale-up. We identified 90 population health interventions targeting physical activity, nutrition or obesity-related health behaviours and collected publicly available information on their key characteristics and outcomes. We then assessed the differences between interventions that followed a research pathway and those that did not. Two-thirds (n = 60) of the interventions were reported as having followed a research pathway. Univariate logistic regression analysis revealed that these interventions were more likely to be health education interventions [odds ratio (OR): 5.56; 95% confidence interval (CI): 1.38-22.38], developed by research institutes (OR: 12.81; 95% CI: 3.47-47.34), delivered in North America (OR: 4.13; 95% CI: 1.61-10.62), and less likely to be owned (OR: 0.35; 95% CI: 0.14-0.88) or funded by government organizations (OR: 0.37; 95% CI: 0.14-0.95). Interventions that followed a research pathway were nearly three times more likely to have a positive impact on population health (OR: 2.72; 95% CI: 1.04-7.14). Interventions that followed a research pathway to scale-up were no more likely to be sustained longer than those that did not. Differences exist across real-world interventions between those that follow a research pathway to population-scale delivery and those that do not, regarding organizational and environmental context. A key benefit of research pathway to scale-up is the impact it has on health outcomes.

Upgrading hazelnut skins: Green extraction of polyphenols from lab to semi-industrial scale.

Hazelnut skins (HS) are usually managed as waste; however, this by-product is a source of bioactive compounds, with potential applications in feed and food sectors. Phenolic compounds can be extracted using green protocols combining enabling technologies and green solvents. This work investigates subcritical water extraction (SWE) of bioactive compounds from HS. A laboratory-scale study was performed on four different batches, with significant batch-to-batch heterogeneity. The evaluation of polyphenolic profiles and antioxidant activities afforded promising results compared to the benchmark of reflux maceration. To evaluate process effectiveness, the extraction protocol was replicated on a semi-industrial plant that processed 8 kg of matrix. Downstream processes have been optimized for scale-up, demonstrating the effectiveness of SWE in retaining product concentration and bioactivity avoiding excipients in spray-drying phase. Hazelnut extracts exhibited antibacterial properties against animal- and food-borne pathogens, supporting their potential use as sustainable feed ingredients for improved hazelnut production and animal farming practices.

The pathway to delivering injectable CAB for HIV prevention: strategies from global PrEP leaders leveraging an adapted version of the Intervention Scalability Assessment Tool (ISAT).

BACKGROUND: Longer-acting cabotegravir (CAB) is a novel, safe, and efficacious pre-exposure prophylaxis (PrEP) for HIV prevention. As we near a time for CAB scale-up, the experience of global leaders in PrEP research and implementation can be leveraged to identify optimal strategies for scaling and integrating CAB into existing PrEP infrastructure worldwide. METHODS: We recruited leaders of HIV prevention clinical trials and large PrEP programs through a combination of purposive and snowball sampling for participation in individual interviews. We conducted interviews using a semi-structured guide that compared CAB to oral PrEP and sought perspectives on barriers and strategies for CAB scale-up. Interviews were conducted virtually, audio recorded, and transcribed. We used thematic analysis, grounded in an adapted version of the Intervention Scalability Assessment Tool (ISAT), to identify critical elements for optimizing delivery of CAB. RESULTS: From October 2021 to April 2022, we interviewed 30 participants with extensive experience in PrEP research, care, and programming. Participants worked in all seven WHO regions and reported a median of 20 years working in HIV and 10 years in PrEP. Participants agreed that CAB was efficacious and discrete, therefore having the potential to address current concerns about oral PrEP adherence and stigma. Participants indicated direct and indirect costs for provider training, expansion of existing medical infrastructure, and the current medication cost of CAB as major concerns for roll out. The true cost to the end-user and health system were unknown. There were some conflicting strategies on how to best address product targeting, presentation of efficacy, and timing of product availability with scale-up. Some thought that targeting CAB for the general population could normalize PrEP and decrease stigma, while others thought that prioritizing key populations could optimize impact by targeting those with highest risk. Overall, participants emphasized that to ensure successful CAB scale-up, communities and stakeholders must be involved at every stage of planning and implementation. CONCLUSIONS: Our evaluation found that although there is a clear and urgent need for additional HIV PrEP options beyond daily oral PrEP, CAB scale-up must be thoughtful, flexible, and based in lessons learned from oral PrEP rollout.

Further Exploring the Public Health Implications of the Network Scale-Up Method: Cross-Sectional Survey Study.

Background: The decline in the number of new HIV infections among adults has slowed down, gradually becoming the biggest obstacle to achieving the 2030 target of ending the HIV/AIDS epidemic. Thus, a political declaration to ensure that 90% of people at high risk of HIV infection can access comprehensive prevention services was proposed by the United Nations General Assembly. Therefore, obtaining an accurate estimated size of high-risk populations is required as a prior condition to plan and implement HIV prevention services. The network scale-up method (NSUM) was recommended by the United Nations Programme on HIV/AIDS and the World Health Organization to estimate the sizes of populations at high risk of HIV infection; however, we found that the NSUM also revealed underlying population characteristics of female sex workers in addition to being used to estimate the population size. Such information on underlying population characteristics is very useful in improving the planning and implementation of HIV prevention services. This is especially relevant for people who inject drugs, where in addition to stigma and discrimination, criminalization further hinders access to HIV prevention services. Objective: We aimed to conduct a further exploration of the public health implications of the NSUM by using it to estimate the population size, popularity ratio, and information transmission rate among people who inject drugs. Methods: A stratified 2-stage cluster survey of the general population and a respondent-driven sampling survey of people who inject drugs were conducted in the urban district of Taiyuan, China, in 2021. Results: The estimated size of the population of people who inject drugs in Taiyuan was 1241.9 (95% CI 1009.2-1474.9), corresponding to 4.4×10-2% (95% CI 3.6×10-2% to 5.2×10-2%) of the adult population aged 15-64 years. The estimated popularity ratio of people who inject drugs was 53.6% (95% CI 47.2%-60.1%), and the estimated information transmission rate was 87.9% (95% CI 86.5%-89.3%). Conclusions: In addition to being used to estimate the size of the population of people who inject drugs, the NSUM revealed that they have smaller-sized personal social networks while concealing their drug use, and these underlying population characteristics are extremely useful for planning appropriate service delivery approaches with the fewest barriers for people who inject drugs to access HIV prevention services. Therefore, more cost-effectiveness brings new public health implications for the NSUM, which makes it even more promising for its application.

Optimized batch cultivation and scale-up of Bacillus thuringiensis for high-yield production of poly(3-hydroxybutyrate-co-3-hydroxyvalerate).

Addition of statistically optimized concentration of electron acceptor, propionic acid (1.2 g/L) at different cultivation times (0 h, 14.86 h and 19 h) during batch cultivation of B. thuringiensis in mixed substrate (glucose and glycerol) featured production of 8 g/L of biomass and 3.57 g/L of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) containing 0.805 g/L of 3-hydroxyvalerate concentration. Successful scale up of batch cultivation from 7 L to a 70 L bioreactor was, thereafter, achieved using power/volume (P/V) criteria with maximum PHBV and biomass concentration of 3.57 g/L and 7.15 g/L respectively. Characterization of PHBV so produced was carried out using NMR, FTIR, DSC and TGA to elucidate its structure, thermal properties and stability to map their applications in society. These findings highlight the potential of the optimized batch cultivation and scale-up process in producing PHBV emphasizing its relevance in sustainable biopolymer production.

Genetic heterogeneity of engineered Escherichia coli Nissle 1917 strains during scale-up simulation.

Advanced microbiome therapeutics have emerged as a powerful approach for the treatment of numerous diseases. While the genetic instability of genetically engineered microorganisms is a well-known challenge in the scale-up of biomanufacturing processes, it has not yet been investigated for advanced microbiome therapeutics. Here, the evolution of engineered Escherichia coli Nissle 1917 strains producing Interleukin 2 and Aldafermin were investigated in two strain backgrounds with and without the three error-prone DNA polymerases polB, dinB, and umuDC, which contribute to the mutation rate of the host strain. Whole genome short-read sequencing revealed the genetic instability of the pMUT-based production plasmid after serial passaging for approximately 150 generations using an automated platform for high-throughput microbial evolution in five independent lineages for six distinct strains. While a reduction of the number of mutations of 12%-43% could be observed after the deletion of the error-prone DNA polymerases, the interruption of production-relevant genes could not be prevented, highlighting the need for additional strategies to improve the stability of advanced microbiome therapeutics.

Industry's Perspective on Challenges Assessing the in Vivo Impact of Removing Titanium Dioxide (TiO2) from Drug Products.

The European Commission (EC) has tasked the European Medicines Agency (EMA) to provide a recommendation towards the acceptability of titanium dioxide (TiO2) in pharmaceutical products by early 2024 to inform on final decision in early 2025[1]. Unlike the already implemented ban of TiO2 in foods, removing this excipient from pharmaceutical products will likely have significant impact on the pharmaceutical industry, regulatory agencies, and patients. This commentary explores the challenges facing the pharmaceutical industry tasked with supporting the development and registration of TiO2 free (TF) drug products. Specifically, justification of formulation changes and potential impact to in vitro and in vivo performance, as well as differences in global regulatory comparative dissolution requirements to justify changing to TF drug product are discussed. Particularly, the uncertainties around how a formulation change such as removal of TiO2 from immediate release solid oral dosage forms will be viewed in Europe compared to other regions is discussed. To respond to these challenges and avoid disruption to the medicines supply chain in case in vitro data such as dissolution is either too challenging or insufficient to justify changing to TF product, pharmaceutical companies may have to decide if the level of risk is worth the effort needed to reformulate, develop, and register a new TF product.