Process analytical technology (PAT) is a framework that encourages the implementation of analytical tools for real-time monitoring of critical batch components during the production of pharmaceuticals. In mammalian cell cultures, Raman spectroscopy as a PAT tool has demonstrated its value for accurate in-line monitoring of various process parameters as well as quality attributes. However, as process development cultures are transferred to manufacturing scales, the small-scale models are often adjusted either with new calibration data or with transfer learning algorithms. This model adjustment step makes the overall process transfer to manufacturing less straightforward from an analytical standpoint, requiring additional resources and specific data treatment expertise to fine tune the models. In this work, we compare a specific spectra processing approach against existing approaches reported in the literature for processing Raman spectra in the context of scale-up challenges. The proposed novelty is a preprocessing strategy which computes spectra normalization based on the 3100-3700 cm-1 band. Data generated at 3 L scale were used to monitor a 3000 L bioreactor without model recalibration or transfer learning with equivalent accuracy as a same scale 3 L bioreactor monitoring. While broader validation case would be warranted, these results demonstrate a direct scale-up model implementation in large scale manufacturing under the studied conditions.
This work presents a methodology for the generation of continuous fibre trajectories based on principal stress directions in continuous fibre-reinforced additive manufacturing (CFAM). The material system considered consists of continuous carbon fibre (CCF-1.5K) embedded in a CFC-PA thermoplastic matrix. CFAM enables the deposition of fibres along tailored paths, allowing improved alignment with the load direction, compared to traditional composite manufacturing. In this way, the strong anisotropy of composite materials, typically considered a limitation, is exploited as a design opportunity by aligning fibres with the structural load paths. The proposed approach combines finite element analysis with a path generation procedure, including the computation of principal stress directions, the extraction of streamlines of the principal stress field, and a dedicated post-processing stage aimed at obtaining continuous and manufacturable fibre layouts. The effectiveness of the method is assessed through a finite element-based comparison with conventional fibre configurations, showing an increase in global stiffness of approximately 20% with respect to the best-performing unidirectional layout. In addition, the feasibility of the generated trajectories is demonstrated through printing tests performed on a continuous fibre additive manufacturing system. The results confirm that the proposed methodology enables the generation of physically realizable fibre paths while improving structural performance.
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Venipuncture is a fundamental clinical skill that requires repeated practice in a safe learning environment before being performed on patients. However, commercial venipuncture simulators are often expensive and not always available in sufficient numbers for repeated student training. This study aimed to design, manufacture, and evaluate a low-cost, reusable venipuncture training model for medical education. The simulator was developed using a silicone-based soft-tissue model with embedded silicone tubing simulating superficial veins at different depths. A passive hydrostatic pressure system was used to provide visual feedback during successful venipuncture. The model was manufactured using a 3D-printed mold, platinum-cure silicone, silicone tubing, and a porous support layer to improve durability and fluid absorption. The total material cost per model was approximately 4-5 EUR. The model was used in a practical training session with 50 medical students as part of a scheduled educational activity. Following the training, participants were invited to complete a voluntary and anonymous questionnaire designed to evaluate the perceived realism, usability, and educational value of the model. The questionnaire consisted of 10 items rated on a 5-point Likert scale. The instrument was developed specifically for this study and was not previously validated; therefore, the results should be interpreted as exploratory measures of user perception. Students reported high satisfaction across all evaluated categories, particularly ease of use, the ability to perform repeated venipuncture, and the visual feedback system. This low-cost, in-house manufactured venipuncture simulator provides a practical and sustainable solution for procedural training. Its affordability, reusability, and ease of manufacturing make it suitable for widespread use in medical education, particularly in settings with limited access to commercial simulation equipment.
Balancing the strength and toughness of materials remains a long-standing core challenge in materials science. In this study, a novel design strategy for Al2O3/Mg interpenetrating phase composites (IPCs) is proposed. Al2O3 ceramic scaffolds with two types of Split-P triply periodic minimal surface (TPMS) structures (shell-type and solid-type) were fabricated via stereolithography (SLA) and then infiltrated with AZ91D magnesium alloy using lost foam casting (LFC), and consequently, Al2O3/Mg IPCs were successfully fabricated. In the proposed method, cocontinuous interpenetration of ceramic and metal phases is achieved, ultimately forming a dense interlocking structure with no interfacial delamination and a relative density exceeding 95%. Quasi-static compression tests and finite element simulations confirm that the as-built composites exhibit a compressive strength of up to 187.5 MPa and a specific energy absorption of 11.18 J/g; among them, the solid-type IPC shows a slightly higher compressive strength, while the shell-type IPC demonstrates superior energy absorption performance. The excellent performance of the IPCs is attributed to the TPMS structure that effectively mitigates the local stress concentration prone to occur in traditional truss structures, along with the synergistic strengthening effect between ceramic and metal phases that substantially improves the plastic deformation capacity and energy dissipation efficiency of the composites. This study provides a new idea for the design and fabrication of high-strength and lightweight composites, which hold significant application potential in lightweight load-bearing and impact-resistant energy-absorbing fields.
Recent clinical trials have highlighted the potential of P63+ lung progenitor cell (LPC) transplantation for lung repair and regeneration. Currently, ex vivo P63+ LPC expansion depends on coculture with growth-arrested fibroblast feeder cells (GAFs), necessitating repeated purification during passaging. While differential enzymatic digestion (DED) and fluorescence- or magnetic-activated cell sorting techniques (FACS/MACS) offer partial solutions, a scalable, efficient, and consistent separation technique remains unmet, particularly for cell therapy manufacturing. Here, we present a multidimensional double spiral (MDDS) inertial microfluidic device designed for high-throughput label-free enrichment of P63+ LPCs. The MDDS device achieves cell size-based separation of P63+ LPCs and growth-arrested feeder cells, processing at a rate of 106 to 107 cells per minute. MDDS-sorted P63+ LPC purity correlates with the LPC-to-GAF ratio in culture. With an initial ratio >1:1, it yields P63+ LPC purity exceeding 80%. Moreover, the device consistently recovers >80% of P63+ LPCs, with the unrecovered fraction enriched in senescent cells exhibiting compromised clonogenicity and differentiation capacity. In direct benchmarking against DED and FACS, the MDDS device delivered a balanced performance in terms of purity and recovery, while offering advantages in throughput, consistency, and scalability. We propose that this technology could enable more consistent and efficient enrichment of feeder-cultured P63+ LPCs, thereby supporting more robust clinical manufacturing processes.
The microstructure of high-strength martensitic steel specifically made for additive manufacturing was modified via in situ plasma arc remelting (PAR) to improve its surface properties. The results reveal that the microstructure is characterized by the intragranular martensite and intergranular eutectic structure of high-strength martensitic steel. The intragranular worm-like δ-ferrite embedding in the martensite matrix was clearly observed after PAR. Compared with the as-deposited part, the tensile strength of the PAR part reached 1753 MPa, and the ductility increased to 2.3%. The strength and elongation had increased by 20% and 229%, respectively. After in situ PAR, the wear loss decreased to 80% of the tailored high-strength martensitic steel, and the corrosion current density decreased to 17%. Both the as-deposited part and the PAR part exhibited significant intergranular corrosion morphological characteristics.
Adoptive cell therapy has witnessed significant progress with the success of chimeric antigen receptor (CAR) T cells for treating cancer. However, their autologous nature limits scalability, and increases production time and manufacturing costs. Additionally, CAR-T cell administration can result in severe toxicities, including cytokine release syndrome (CRS) and neurotoxicity. To address these issues, allogeneic, natural killer (NK) cells are being explored as an alternative. NK cells are cytotoxic lymphocytes that play a pivotal role in tumor surveillance and eradication. Unlike T cells, NK cells can identify and eliminate targets without MHC restriction or prior sensitization. Furthermore, NK cells exhibit enhanced responses after exposure to virus infections or cytokine activation (cytokine induced memory-like). Allogeneic NK cell therapies offer a promising alternative to autologous cell therapies, with reduced risk of graft-versus-host disease and rapid availability. This review summarizes the current landscape of allogeneic memory-like NK cell therapies, including clinical applications and challenges.
Natural killer (NK) cells are promising platforms for off-the-shelf immunotherapy, yet nonviral precision engineering remains limited by poor HDR efficiency, DNA toxicity, and manufacturing challenges. The aim of this study was to establish a high-yield, nonviral knock-in platform. Through extensive in-depth rational screens, we achieved ∼90% HDR insertion of therapeutic payloads while maintaining 100% postediting recovery. By hijacking endogenous transcriptional programs, we installed genetic circuits into defined genomic loci to tune transgene expression. To enable context-dependent therapeutic responses, we integrated a synthetic positive feedback circuit at the CISH locus, which enhanced NK cell persistence and drove strong expression of anti-CD22/19 dual CAR. A hypoxia-responsive IL-12 circuit gated by the PFKFB4 promoter restored cytotoxicity under environmental stress. Finally, we showed this platform is compatible with GMP manufacturing and supports clinical-scale expansion. These findings provide a scalable framework for programmable, nonviral editing of NK cell effector functions for therapeutic and research applications.
Indigo, a ubiquitous vat dye in denim manufacturing, is characterized by its exceptionally low aqueous solubility, necessitating chemical reduction to its leuco form using hazardous agents such as sodium dithionite. This conventional process yields sulfite, sulfate, and sulfide byproducts, leading to significant environmental and toxicological concerns. To address these limitations, this study synthesized 1:1 molar ratio inclusion complexes of indigo with β-cyclodextrin (β-CD) and hydroxypropyl-β-cyclodextrin (HP-β-CD) to enhance solubility and stability without chemical intervention. The formation of these complexes was rigorously validated through UV-Vis, FT-IR, PXRD, TEM, and 1H-NMR spectroscopy. Notably, the aqueous solubility of indigo was enhanced 3.58-fold and 5.55-fold for the β-CD and HP-β-CD complexes, respectively. Furthermore, both complexes demonstrated superior thermal and photostability, with HP-β-CD exhibiting the most pronounced effects. This cyclodextrin-assisted solubilization was successfully extended to naturally extracted Jeju Indigo, underscoring its broad applicability. Our findings suggest that cyclodextrin-based encapsulation offers a sustainable and effective alternative to conventional reduction-oxidation dyeing processes.
Inflammation is a complex and highly regulated defensive response of the body to injury, infection, or abnormal stimuli (such as pathogens, toxins, and physical/chemical damage). Natural anti-inflammatory drugs hold significant potential in the pharmaceutical field due to their multi-target effects, high safety profiles, and low toxicity. For example, EGCG can inhibit the phosphorylation of p38 and JNK, thereby reducing the activation of the AP-1 transcription factor and subsequently downregulating the expression of inflammatory genes. Luteolin inhibits inflammasome assembly by blocking potassium ion efflux, suppressing mitochondrial reactive oxygen species generation, or directly binding to NLRP3. Over the past decade, extensive research has been conducted on their physicochemical properties and dosage forms, leading to the development of various natural anti-inflammatory drug formulations using both traditional and modern technologies. Furthermore, the combination of these natural anti-inflammatory agents with other drugs can further expand their therapeutic applications. Meanwhile, emerging technologies such as 3D printing and AI-assisted design have demonstrated significant potential for application in formulation development. Despite these advancements, the current research field still faces critical challenges: the use of toxic excipients in certain formulations poses biosafety risks (for example, glutaraldehyde offers advantages such as high cross-linking efficiency, strong cross-linking strength, and mature manufacturing processes, but it also exhibits high toxicity and biosafety deficiencies); and the translation of research findings on natural anti-inflammatory drugs into commercial products remains insufficient (due to challenges in large-scale production, storage difficulties, and regulatory standards). Through this review, we hope to draw more attention to the development potential of natural anti-inflammatory drugs and the aforementioned issues, as well as to offer some foresight for the exploration and development of other natural products.
The electrocatalytic chlorine evolution reaction (CER) is essential to modern chlor-alkali industry, yet conventional RuO2 catalysts suffer from parasitic oxygen evolution. High-entropy ruthenium oxides (Ru-HEO) are promising alternatives, but their practical design is hindered by complex composition-structure-performance relationship. Herein, we construct a Pareto-guided multi-objective Bayesian optimization framework to enable autonomous high-throughput exploration of quinary Ru-HEO system. Through this trade-off strategy, we identify compositions that efficiently balance mass activity, Cl2 selectivity and material cost. The leading Ru-HEO catalyst with only 8.4 at% Ru achieves a remarkable activity of 5083 A g-1 Ru at 1.50 V versus RHE and maintains excellent 100-h stability, outperforming commercial RuO2 and the state-of-the-art catalysts reported. Integrated into a photovoltaic-electrochemical (PV-EC) prototype device and tested under simulated diurnal illumination, it sustains >95% selectivity, a maximum solar-to-chemical (STC) efficiency of 14.6% and projected Cl2 production costs as low as $0.177 per kg. Our work establishes a closed-loop, AI-accelerated research paradigm that integrates multi-objective optimization with robotic experimentation, offering a generalizable and expedited pathway toward high-performance electrocatalysts for sustainable chemicals manufacturing.
Human induced pluripotent stem cells (iPSCs) are a promising starting material that can be differentiated into therapeutic cell products for clinical trials. The production of clinically applicable drug products requires good manufacturing practice (GMP)-compliant iPSCs as the initial starting material. Under the framework of the Korean Ministry of Food and Drug Safety, we generated a master cell bank (MCB) of a human iPSC line, KNIH01-MCB, from peripheral blood mononuclear cells obtained from a healthy female donor under a GMP system. To verify cell identity and quality, we performed a risk assessment of genetic stability, pluripotency, cell line identity, and sterility. For this purpose, we used validated methods, including polymerase chain reaction, karyotyping, short tandem repeat analysis, ABO/human leukocyte antigen (HLA) typing, and bacterial growth testing. After material qualification, the cells underwent comprehensive quality testing, including viral screening, ABO/HLA typing, and sterility testing. We successfully established an MCB of the iPSC line, enabling clinical readiness, including full traceability and donor eligibility for clinical applications. The MCB met the quality control criteria of our in-house GMP system. These iPSCs provide a GMP-compliant and clinically applicable starting material for therapeutic cell manufacturing intended for human use. Induced pluripotent stem cells (iPSCs) are cells that can develop into almost any cell type in the body, making them a valuable tool for developing new medical treatments. In this study, we generated good manufacturing practice-compliant iPSCs from the blood cells of a healthy female donor using a non-integrating episomal vector, which avoids permanent changes to the cells’ DNA. We performed extensive testing to confirm that the reprogramming vector was fully cleared and that the cells remained genetically stable and free of contamination. We also verified their identity and quality using multiple laboratory methods. These results support the use of our iPSCs as a reliable and safe starting material for future clinical applications. These cells can be used to produce various differentiated cells, including hematopoietic cells such as red blood cells and platelets, for clinical use.
Regulatory reliance pathways are increasingly being recognized as essential tools to accelerate patient access to medicines globally while building National Regulatory Authorities (NRA) capacity and capability. Although significant progress has been made in a number of international pilots for Chemistry, Manufacturing and Controls (CMC) post approval changes, there has been more limited application to labelling variations including indication extensions. This paper describes the first reliance pilot for an indication extension, which engaged 21 NRAs across multiple regions. The pilot leveraged the European Medicines Agency (EMA) as a reference agency and incorporated a digital platform, whose use was optional by the NRAs involved, for real-time sharing of questions and responses. The objectives were to reduce approval timelines, promote regulatory convergence, streamline health authority questions and enhance transparency among participating authorities. Results demonstrated significant reduction in approval timelines, with an average reduction of 3 months per country compared to standard timelines across participating countries. This article outlines the process of establishing the pilot, including planning and engagement with regulatory authorities, analysing success factors and challenges encountered including recommendations for optimising the process further. The findings suggest that regulatory reliance for indication extensions can substantially improve efficiency in regulatory processes, reducing approval timelines and ultimately benefiting a wider patient population to access medicines globally and timely.
Aromatic hydrocarbons such as benzene, toluene, and ethylbenzene are extensively used as solvents in coatings, resin, and artificial leather industries. Azeotropic mixtures involving these compounds are commonly encountered in chemical manufacturing, where accurate azeotropic temperature and composition are essential for designing and optimizing separation processes such as extractive and pressure-swing distillation. In this study, two quantitative structure-property relationship (QSPR) models were developed to predict the azeotropic temperature and composition of binary mixtures containing aromatic hydrocarbons using only molecular structural information. The models show excellent agreement with experimental data (R2 = 0.9454 and 0.9448, R adj 2 = 0.9400 and 0.9413). Internal validation via leave-one-out cross-validation yields R cv 2 = 0.9308 and 0.9364, while external validation using an independent test set yields Q ext 2 = 0.8939 and 0.9364, indicating strong robustness and superior predictive performance compared to previously reported models. Molecular geometries were optimized using HyperChem 8.0, employing MM + and PM3 methods. Molecular descriptors were calculated using the Online Chemical Modeling Environment (OCHEM). Binary mixture descriptors were derived from pure-component descriptors via Kay's mixing rule. The genetic function approximation (GFA) algorithm was used to select the most relevant descriptors, and predictive models were constructed using multiple linear regression (MLR). Model robustness and predictive capacity were evaluated using leave-one-out cross-validation and an external test set, with applicability domains assessed via Williams plots. All computational procedures and modeling analyses were performed using OCHEM, SPSS, and HyperChem 8.0.
Since the Fukushima NPP accident in Japan in 2011, negative rumours about radioactivity and radiation in general have been spreading in Korea for more than 12 consecutive years, causing various social conflicts as well as actual physical losses. Industry stakeholders have suffered significant losses due to the collapse of fisheries, the voluntary short-term closure of schools, denuclearisation policies, road asphalt restoration projects, a bed manufacturing business collapse, the closure of seawater desalination facilities, thyroid cancer lawsuits, etc. The scientific basis for the spread of negative radioactivity rumours in Korea all contain radiological levels far below 100 mSv. This is all related to the interpretation and application of the LNT model. Since the losses due to radiation-related rumours after the Fukushima NPP accident are based on the ICRP recommendations, the relationship between legal effects and losses should be sufficiently addressed to provide criteria for preventing future losses. The use of radiation is related to several fundamental human rights under the Constitution. Therefore, it is necessary to consider whether the ICRP recommendations comprehensively reflect the balance of these constitutional rights.
Glucagon-like peptide-1 (GLP-1), a 31-amino acid incretin hormone, is widely used in the treatment of type 2 diabetes mellitus due to its glucose-dependent insulinotropic activity. However, its small size makes it highly prone to proteolytic degradation in microbial expression systems such as Escherichia coli, leading to reduced manufacturing yield. While fusion to cleavable protein tags can improve peptide stability during purification, excessively large tags often compromise the overall yield, especially when the target peptide is smaller than the fusion partner. To overcome this limitation, we have engineered 11 cleavable fusion tag constructs (LP1-LP11) for recombinant expression of Arg34-GLP-1(7-37) (liraglutide precursor) in E. coli. The 11 constructs differed only in the tags. The expression vector contained a T7 leader sequence, affinity tags (6×His/6×Arg), inclusion body tags (11-125 amino acids), and TEV protease cleavage sites. Among the 11 tags, LP8 with a compact 4.0 kDa tag achieved the highest expression, yielding 133 mg/L of fusion protein and a calculated liraglutide precursor yield of 60 mg/L based on mass fraction (45% of fusion mass), with an actual recovered yield of 14.6 mg/L after RP-HPLC purification, largely due to efficient inclusion body formation (>95% insolubility) and enhanced translational initiation driven by the T7 leader sequence. The purified peptide's identity and sequence integrity were confirmed by LC/MS analysis. The primary advantage of this approach is mass fraction optimization which focuses on minimizing fusion-tag mass to maximize yield relative to the tag size without compromising inclusion-body formation thereby providing a scalable and economical approach for GLP-1 analogs and potentially other peptide-based biopharmaceuticals.
Three-dimensional integrated circuits (3D ICs) have emerged as a key technology to sustain scaling trends in the microelectronics industry. This advancement calls for a fundamental shift in how electrical interconnects are implemented, with through-silicon vias (TSVs) playing a pivotal role in enabling vertical connectivity between stacked chips. However, the metallization of TSVs traditionally involves elaborate and demanding processes, which can limit the speed and flexibility of prototyping and design modifications. In this paper, we investigate the use of Ultra-Precise Dispensing (UPD) technology of novel silver nanoparticle-based pastes as a simple and adaptable alternative to the metallization of TSVs process. The TSV filling process is outlined, followed by a detailed analysis of their morphology, filling quality, and electrical performance. We successfully achieve filled vias through a 280 μm thick silicon substrate with diameters down to 20 μm, resulting in an aspect ratio of up to 14:1, exhibiting favorable electrical properties. This work contributes to the achievement of dense, high-aspect ratio TSV fabrication using additive manufacturing, demonstrating a path towards reduced complexity of standard technology processes cycle, lower cost potential, and increased design flexibility.
Monometallic Zn is a promising anode material for post-lithium batteries owing to its high theoretical capacity, low cost and natural abundance. However, the practical application in zinc-air batteries (ZABs) remain limited by uncontrollable dendrite growth and severe voltage polarization during repeated cycling. Herein, we report a self-supported Sb-modified porous lamellar zincophilic electrode (Sb-pZn) as a reversible, dendrite-free and high-performance hostless Zn anode for aqueous alkaline rechargeable ZABs. The porous lamellar framework is fabricated by selectively dissolving the Al component from the eutectic Zn-Al alloy via chemical dealloying, followed by surface modification through galvanic replacement in SbCl3 solution. The Sb-pZn anode with lamellar thickness of ~2920 nm (Sb-pZn-2920) enables continuous electron and ion transport pathways through quasi-periodic lamellar channels and interconnected ligament network, ensuring uniform Zn deposition and dissolution. Moreover, Sb incorporation introduces abundant zincophilic sites that accelerate charge-transfer kinetics and enhance reversibility. As a result, the Sb-pZn-2920 anode shows exceptional Zn plating-stripping behaviours with ultralow overpotential of 31.7 mV for up to 1700 h at 2 mA cm-2 in symmetric cells and maintains stable charge-discharge cycling for 240 cycles at 20 mA cm-2 under ambient conditions in ZAB full cells. This work presents a facile and scalable strategy for constructing high-performance Zn anodes and offers valuable insights into the development of practical, durable and reversible ZABs. This work presents a novel strategy for designing high-performance zinc anodes, providing key insights into achieving highly reversible, stable and practical zinc-air batteries for next-generation energy storage.
Prosthetic components from high-income countries (HICs) are often replaced not because they are broken, but because of guidelines or expired warranties, meaning they may still be usable. As most HICs classify prostheses as single patient multi-use devices, components are often disposed of or donated to low- and middle- income countries (LMICs) where medical device regulatory frameworks are limited or non-existent. A lack of standards guaranteeing the quality of donated prosthetic components could lead to a violation of the World Health Organization's principles of good donation. Here, we work towards the creation of a set of standards by quantifying the efficacy of a second-hand donated prosthetic foot quality checklist developed by STAND. We compared 170 checked to 196 unchecked feet received by prosthetic and orthotic centres in Fort Portal, Uganda, and found checklist implementation increased the percentage of usable feet from 83.16% to 94.12%. Foot brand significantly affected usability, but further data and samples are needed to disentangle the effects of prosthetic foot brand, prosthetic foot model, and centre from which the feet originated on prosthetic foot usability. We propose a rapid and efficient quality assurance checklist as a first step towards a set of standards towards prosthetic foot reuse and discuss future research directions. Research towards the creation of an international set of standards/regulatory requirements governing the use of prosthetic limbs, like the international standards used for prosthetic limb design, would not only enable the safe, useful provision of prosthetic components in LMICs, but would also set the groundwork for understanding how a repair, reuse, and recycle model for prosthetic components might be implemented in HICs. Globally, this could decrease prosthetic provision time, create a circular economy for prosthetic components, and reduce the carbon footprint of prosthetic component manufacture and provision.