Search Results (611)

Background: Regulations in some countries of the world allow self-driving vehicles (private cars and robo-taxis) to operate on geofenced, public roads, yet governments are slow to plan as how best to use this automated technology. We pose research questions about the Australian government’s preparedness, planning gaps for a transition to an autonomous public transport system, and specific system components that require attention. Method: We review the relevant literature, and podcasts of automobile manufacturing experts, and draw on our extensive professional experience advising governments in applying the systems approach to a planning system that includes autonomous transport. Results: Governments must include risk management in Type-II road corridors; develop mobility hubs that connect terminals for fully self-driving vehicles and robo-taxis to connect with public transport systems; and include body corporates when engaging the community in precinct planning. In the discussion, we argue the case for an autonomous urban public transport system where private ownership of vehicles is progressively reduced. Conclusions: Australian governments are not prepared with a systems-wide urban planning process that includes autonomous transport and self-driving vehicles. During the transition period, the existing and new transport systems must operate together, emphasising the leading role for governments. A roadmap for further research and development is outlined and this could provide the framework for urban planning in other jurisdictions. Full article

Background/Objectives: Point-of-care (PoC) manufactured fresh chimeric antigen receptor (CAR)-T cells are typically formulated in hypothermic preservation formulations (HPFs) and stored under hypothermic conditions (2–8 °C) until administered to the patient. However, in current HPFs the shelf life of fresh CAR-T cells is short (~24–36 h) due to limited CAR-T cell stability, which poses significant time constraints on manufacturing procedures and logistics. The objective of this study was to improve the stability and extend the shelf life of fresh clinical-grade CAR-T cell drug products (DPs). Methods: A novel HPF was developed by supplementing a base HPF with the novel excipient SUL-138, which stabilizes mitochondria during hypothermic storage and subsequent rewarming, alone or in combination with endogenous mitochondrial substrates. This panel of HPFs was first screened for their stability-improving characteristics in the model cell line Jurkat cells. Subsequently, HPFs were assessed for their stability-improving characteristics of clinical-grade CD19 CAR-T cell DPs. Critical quality attributes, including CAR-T cell viability, T-cell differentiation state, exhaustion markers, and functional potency were evaluated in a good manufacturing practice (GMP)-compliant stability study up to 72 h. Results: For Jurkat cells, HPFs supplemented with SUL-138 and a combination of glucose, glutamine, and succinate demonstrated the greatest stability improvement at 2–8 °C, improving cell viability from ~1% to >85% after 72 h. For CAR-T cells, supplementation of HPFs with SUL-138 alone demonstrated the greatest improvement, resulting in a CAR-T cell viability from ~40% to >85% after 72 h of storage at 2–8 °C, while no additional benefits from mitochondrial substrates were observed. The novel HPF did not significantly impact CAR-T cell potency test results, T cell subset distribution, or exhaustion markers compared to control. Conclusions: A novel clinical-grade HPF that significantly improved fresh CAR-T cell stability during hypothermic storage was developed. This novel HPF can aid in the establishment of GMP-compliant and PoC CAR-T cell manufacturing platforms. Full article

Chimeric antigen receptor (CAR)-T cell therapy has achieved remarkable clinical success in B cell malignancies. However, its efficacy in solid tumors remains limited, in part due to suboptimal expansion, persistence, and restrained effector function. Strategies that promote durable CAR-T cell fitness are therefore required to overcome these barriers. In this study, we generated HER2-CAR-T cells targeting human breast cancer cells and evaluated the impact of different cytokine supplementation strategies on CAR-T cell phenotype and function. We analyzed gene expression patterns and performed repetitive tumor killing assays to assess the ability of CAR-T cells expanded with IL-2 + IL-7 + IL-15 compared with IL-2 alone to maintain proliferation and cytotoxic function across multiple rounds of tumor cell exposure. Compared with IL-2 alone, supplementation with IL-7 and IL-15 significantly enhanced CAR-T cell expansion, preserved stem cell-like features prior to antigen encounter, and promoted superior proliferative capacity. Moreover, CAR-T cells cultured with IL-7+15 or IL-2+7+15 maintained sustained cytotoxicity and exhibited increased antitumor cytokine production during repeated tumor challenges. Notably, IL-7 and IL-15 supplementation induced a CD57⁺ CAR-T cell population that, unlike the immunosenescent CD57⁺ cells reported previously, retained full proliferative and cytotoxic capacity, with CD57 expression being dynamically downregulated upon antigen stimulation. Collectively, these findings demonstrate that incorporation of IL-7 and IL-15 into CAR-T cell manufacturing protocols substantially improves expansion, persistence, and effector function, supporting their use as a strategy to enhance CAR-T cell performance against solid tumors. Full article

Approximately one quarter of the European Union’s (EU’s) CO₂ emissions originate from the transport sector, of which road transport, such as cars and heavy-duty vehicles, contributes roughly 72%. Moreover, according to the European Automobile Manufacturers’ Association, 92% of cars in the EU are internal combustion engine vehicles powered by fossil fuels. Therefore, boosting the adoption of Electric Vehicles (EVs) is considered one of the most prominent solutions for reducing GHG emissions and achieving the EU’s climate targets. To increase EV adoption and fulfil the demand of EV users, adequate EV Charging Stations (EVCSs) are required. Nevertheless, since most EVCSs are supplied by electricity grids that remain predominantly fossil fuel-based, their operation entails substantial indirect GHG emissions. A prominent approach to reducing grid-related emissions is integrating renewable energy sources (RESs) with EVCSs, thereby lowering emissions and alleviating grid stress. Although promising, the energy, economic, and environmental (3E) benefits of this integration remain insufficiently explored. Therefore, this study develops and applies a 3E optimisation framework to assess the feasibility and performance of RES-powered EVCS at NOVA University Lisbon (UNL). Data was collected from the UNL parking area, such as time of arrival, and time of departure. Also, a rule-based algorithm was developed to curate data and estimate the EVCS load profile. Furthermore, HOMER optimisation software was employed to evaluate four scenarios, including (i) an EVCS based on PV, Wind Turbine (WT), and the grid, (ii) an EVCS based on PV and the grid, (iii) an EVCS based on WT and the grid, and (iv) an EVCS based only on energy withdrawal from the grid (base scenario). Under the adopted techno-economic assumptions, in the most optimised scenario, economic and environmental analyses illustrate significant improvements over the base scenario: CO₂ emissions are five times lower, and cost of energy is significantly lower, resulting in significantly lower EV charging costs for users. The results demonstrate that, through developed feasibility studies, researchers, decision-makers, and stakeholders can reach better conclusions about EVCS planning and management. Full article

Modulation of immune response to target tumor cells has been shown to be a successful strategy for cancer treatment. Over the past years immunotherapy has been integrated into cancer treatment and PD-1 blockers have become the backbone of treatment regimens for multiple cancer types. Several classes of immunotherapies, such as immune checkpoint blockers, bispecific antibodies, chimeric antigen receptor (CAR) T-cells, and tumor-infiltrating lymphocytes (TILs), were approved by the US FDA in the last decade and many more are in clinical trials. Research on redirecting effector T-cells to treat cancer has been aimed at addressing the limited responses in solid tumors, emergence of resistance, treatment-limiting adverse events and logistical challenges. Bispecific immune checkpoint blockers, developed to simplify the combination therapies; bispecific T-cell engagers, developed to connect the effector T-cells with tumor cells; and the next generation of CAR T-cells and the next generation of TIL therapies, developed to improve efficacy in solid tumors, are currently under clinical evaluation. This narrative review aims to summarize the current status of T-cell-directed immunotherapy, describing the brief history of development, clinical success, challenges and latest advancements that are under clinical evaluation. Evolution of monoclonal antibodies to bispecific antibodies and bispecific T-cell engagers, the latest advances in adoptive cell therapies, including the optimization of CAR T-cells for solid tumors, allogenic, universal CAR T-cells and in vivo CAR T-cell therapies are discussed in the review along with the key challenges of the therapies, such as primary and acquired resistance, limited efficacy in solid tumors, manufacturing and logistical challenges, and treatment-related toxicities. Full article

Adoptive immunotherapy using ex-vivo-amplified autologous αβ T cells has achieved notable success in the treatment of diverse cancer types. Pre-eminent among these developments has been the advent of chimeric antigen receptor (CAR) T cell therapy, which has revolutionised the treatment of selected haematological malignancies. However, autologous CAR T cell immunotherapy is poorly scalable and has demonstrated limited efficacy against solid tumours. Accordingly, there has been significant interest in alternative strategies that may bridge these gaps. The use of γδ T cells is an attractive alternative since they possess intrinsic anti-tumour activity and do not elicit graft versus host disease (GvHD) when employed as an allogeneic drug product. In this review, we evaluate the potential use of γδ T cells for cancer immunotherapy and how manufacturing and genetic engineering refinements can be used to potentiate this activity. We also summarise current clinical experience with CAR γδ T cell therapies and discuss the implications of these findings for the next generation of cellular immunotherapies. Full article

Invariant natural killer T (iNKT) cells are a unique lymphocyte subset that bridge innate and adaptive immunity through recognition of glycolipid antigens presented by CD1d. Upon activation by ligands such as α-galactosylceramide (α-GalCer), iNKT cells rapidly secrete cytokines, including IFN-γ and TNF-α, thereby activating dendritic cells, natural killer (NK) cells, and cytotoxic T lymphocytes (CTLs) to promote antitumor immunity. Despite their therapeutic promise, clinical translation has been limited by rapid α-GalCer clearance, induction of iNKT cell anergy following repeated stimulation, and the immunosuppressive tumor microenvironment (TME). Recent advances in lipid-engineered nanoparticle systems offer solutions to these challenges by improving ligand stability, enhancing antigen-presenting cell targeting, and enabling controlled release that sustains Th1-biased activation while reducing anergy. Liposomal and polymer-based nano-formulations enhance bioavailability and promote more durable IFN-γ-mediated responses. In parallel, chimeric antigen receptor (CAR)-engineered iNKT cells provide antigen-specific tumor targeting while preserving intrinsic CD1d-restricted immunomodulatory functions, demonstrating encouraging safety and efficacy in early-phase studies. Combination strategies further strengthen iNKT-based immunotherapy. Integration with chemotherapy, immune checkpoint inhibitors such as anti-PD-1 and anti-CTLA-4, and cytokine support enhances effector activation, counteracts TME-induced suppression, and improves therapeutic outcomes. However, challenges remain, including optimization of dosing, control of off-target immune activation, scalable manufacturing, and long-term safety evaluation. Collectively, the convergence of nanotechnology, CAR engineering, and rational combination approaches establishes iNKT cell-based therapy as a promising next-generation immunotherapeutic strategy. Continued refinement of delivery systems, genetic engineering platforms, and translational protocols may enable durable immune reprogramming and improved clinical outcomes in resistant and immunosuppressive cancers. Full article

Car engine quality control is fundamentally hindered by extremely high-dimensional, noisy, and imbalanced multi-sensor data. To overcome these challenges, this paper proposes an edge-deployable diagnostic and predictive framework. First, a Sparse Autoencoder (SAE) maps over 12,000 distributed manufacturing parameters into a robust latent space to filter instrumentation noise. Second, for defect classification, a Class-Specific Weighted Ensemble (CSWE) tackles extreme class imbalance by aggressively penalizing majority-class bias, improving defect interception recall by 7.72%. Third, for transient performance tracking, an Adaptive Regime-Switching Regression (ARSR) replaces manual phase selection with unsupervised regime routing to dynamically weight local experts, reducing relative prediction error by 12%. Rigorously validated across three diverse public datasets (NASA C-MAPSS, AI4I, SECOM) and a physical H4 engine assembly line, the framework achieves an ultra-low inference latency of $80\pm 3$ ms, practically reducing the engine rework rate by 7.2%. Full article

Imidazole dipeptides (IDPs), including carnosine, anserine, and balenine, are functional food ingredients found in meats. They have been reported to exhibit high antioxidant activity. 2-Oxo-imidazole dipeptides (2-oxo-IDPs) are present in trace amounts in various tissues and show notably higher antioxidant activity compared with IDPs. Trace amounts of 2-oxo-IDPs are also present in commercial IDP reagents, suggesting that they affect the antioxidant activity of IDPs. Trace amounts of 2-oxo-IDPs were detected in IDP reagents from various manufacturers by HPLC. Some reagents with trace amounts of 2-oxo-IDPs exhibited higher antioxidant activity in a DPPH radical-scavenging assay compared with high-purity IDP reagents devoid of 2-oxo-IDPs. Therefore, it is important to use highly purified IDP reagents to measure antioxidant activity accurately. The antioxidant activity of highly purified IDPs and 2-oxocarnosine (2-oxo-Car) was evaluated through their ability to protect protein and DNA from ROS. 2-Oxo-Car markedly inhibited the protein and DNA degradation by ClO⁻ and ONOO⁻ compared with IDPs. Moreover, 2-oxo-Car was not cytotoxic, even at high concentrations, and suppressed pyocyanin-induced ROS generation in C2C12 cells compared with IDPs and glutathione. Overall, 2-oxo-IDPs are effective antioxidants and are equivalent or superior to known water-soluble antioxidants, such as glutathione and vitamin C. Full article

Ex vivo chimeric antigen receptor (CAR) T cell therapies have achieved remarkable clinical success over the past decade, enabling effective treatment of several hematologic malignancies once considered incurable. However, their broader use remains limited. Barriers include complex and costly manufacturing, long production timelines, and risk of significant side effects and toxicities, challenges that have been further exacerbated by the reduced investment across the biotech sector since 2022. Emerging in vivo CAR-T approaches seek to overcome many of these limitations by generating CAR-T cells directly within the patient, most commonly using lentiviral or lipid nanoparticles (LNPs) delivery vectors. This strategy has the potential to streamline production, allow more tunable and repeatable dosing, and markedly reduce overall costs. However, it also raises new questions regarding genomic safety, the specificity and durability of CAR expression, host immune responses, pharmacokinetics, and regulatory oversight. In this review, we summarize the major and emerging in vivo CAR-T delivery platforms—analyzing their underlying technology, preclinical and clinical performance, and developmental paths—and discuss the scientific, technical, and biological challenges shaping this rapidly emerging field. We further outline future directions and opportunities in the field of programmable T cell immunity. Full article

The clinical success of chimeric antigen receptor (CAR) T-cell therapies has revolutionized oncology, yet the high costs and logistical complexities of ex vivo manufacturing remain significant barriers to global patient access. In vivo cell therapy, which involves the direct injection of lentiviral vectors (LVVs) to engineer cells within the patient’s body, offers a promising, cost-effective alternative. However, transitioning from ex vivo to in vivo applications necessitates a fundamental shift in LVV biomanufacturing to ensure safety and efficacy. This paper examines the critical bottlenecks in the current LVV production landscape. In upstream processing, we explore LVV particle assembly and maturation mechanisms, the effect of transgene size on LVV functional titers and the formation of non-functional byproducts, including empty and partially formed LVV particles and extracellular vesicles (EVs). These impurities pose severe risks of immunotoxicity and insertional mutagenesis when delivered in vivo. In downstream processing, we highlight the challenges of purifying labile LVV particles, emphasizing the need for rapid, high-resolution separation techniques like continuous processing to maintain functional titers. Furthermore, we address the limitations of current analytical assays, which often fail to distinguish mature, functional LVVs from structurally similar but inactive contaminants. We conclude that the future of in vivo lentiviral therapy depends on developing novel purification strategies based on subtle biophysical differences—such as surface charge and capsid morphology—and implementing robust, high-throughput analytics to ensure delivery of high-purity, potent therapeutic viral vectors. Full article

This paper presents a robust multistage optimization framework for the integration of composite laminates into the car body shell of a low-floor light rail vehicle (LRV). While structural design in low-floor vehicles is typically complex, this methodology successfully balances both static and dynamic requirements through a sequential optimization process. Developed in strict accordance with reference European standards, the methodology addresses the structural challenges inherent in low-floor architectures, where complex load paths and redistributed equipment masses require targeted reinforcement. The proposed approach sequentially addresses dynamic and static requirements through a structural optimization process. Two distinct 10-ply laminate configurations, one symmetric and one asymmetric, were investigated. The results demonstrate that the multistage optimization successfully converged to a highly mass-efficient solution, achieving a 66% reduction in laminate thickness compared to the baseline design. This significant result was accomplished while maintaining full regulatory compliance; the failure index increased by approximately 22.5% and 23.3% for the two composite laminate configurations, respectively, effectively maximizing material utilization. A key finding of this study is the preservation of structural dynamic integrity; the fundamental natural frequency was maintained at approximately 16 Hz, with a high correlation across the first ten vibration modes, confirming that the global dynamic behaviour remains unaffected. These observations provide critical insights into the synergy between hybridization and structural constraints, suggesting a systematic pathway for designers to achieve an optimal trade-off between manufacturing costs, weight reduction, and performance in advanced urban transit platforms. Full article

Lentiviral transduction remains the gold standard in adoptive modified cellular therapy, such as CAR-T; however, genome integration is not always desirable, such as when treating non-fatal autoimmune disease or for additional editing steps using CRISPR to produce allogeneic CAR-modified cells. Delivering in vitro-transcribed (IVT) mRNA represents an alternative solution but the labile nature of mRNA has led to efforts to improve half-life and translation efficiencies using a range of approaches including chemical and structural modifications. In this study, we explore the role of N⁶–methyladenosine (m6A) in a CD19-CAR sequence when delivered to T cells as an IVT mRNA. In silico analysis predicted the presence of four m6A consensus (DRACH) motifs in the CAR coding sequence and treating T cells with an inhibitor of the m6A methyltransferase (METTL3) resulted in a significant reduction in CAR protein expression. RNA analysis confirmed m6A bases at three of the predicted sites, indicating that the modification occurs independently of nuclear transcription. Synonymous mutation of the DRACH sites reduced the levels of CAR protein from 15 to >50% depending on the T cell donor. We also tested a panel of CAR transcripts with different UTRs, some containing m6A consensus motifs, and identified those which further improved protein expression. Furthermore, we found that the methylation of consensus m6A sites seems to be somewhat sequence-context-dependent. These findings demonstrate the importance of the m6A modification in stabilising and enhancing expression from IVT-derived mRNA and that this occurs within the cell, meaning targeted in vitro chemical modification during mRNA manufacturing may not be necessary. Full article

The manufacturing process contributes significantly to the proliferation, metabolic state, and functional persistence of chimeric antigen receptor (CAR)-T cells. However, how different culture systems regulate CAR-T cell metabolism and thereby influence their long-term antitumor activity remains poorly understood. In this study, we compared dynamic cultivation using a wave bioreactor with static expansion systems (gas-permeable and conventional T-flasks) for the production of CD99-specific CAR-T cells. CAR-T cells expanded by the wave bioreactor exhibited faster proliferation and stronger cytotoxicity during culture. Upon repeated antigen stimulation, they retained these enhanced functional properties and showed the reduced expression of immune checkpoint molecules, preferentially preserved memory-like subsets, and displayed transcriptional features consistent with memory maintenance and exhaustion resistance. Targeted metabolomic profiling revealed enhanced Tricarboxylic Acid (TCA) cycle activity and features consistent with sustained oxidative phosphorylation, supporting mitochondrial-centered metabolic reprogramming. In a Ewing sarcoma xenograft model, wave bioreactor-cultured CAR-T cells showed a greater percentage of memory-like tumor-infiltrating lymphocytes. Collectively, these results indicate that wave bioreactor-based dynamic cultivation promotes mitochondrial metabolic reprogramming, which is characterized by an enhanced TCA cycle and sustained oxidative phosphorylation, thereby sustaining CAR-T cell functionality and providing a robust platform for the manufacturing of potent and durable cellular therapeutics. Full article

Smart charging management systems for electric vehicles (SCMSs) enable the effective management of electric vehicle (EV) charging processes using smart technologies. Numerous SCMS technologies have been available for different stakeholders, e.g., EV drivers, charging station managers, and car manufacturers. Despite the ever-increasing interest in SCMSs, the literature lacks in reusable, standardised architecture design that reduces the effort for the development of quality SCMSs. In this paper, we propose a reference architecture (RA) for SCMSs. Our RA design is based on our comprehensive domain analysis that encompasses the analysis of the existing literature and commercial technologies which have been supported by our survey on EV drivers. In our RA, we provide four different viewpoints. The context viewpoint classifies the potential stakeholders and their roles and responsibilities. The module viewpoint defines the software implementation units and their modules that can be used for implementing any SCMSs. The component and connector viewpoint defines the executing parts of any SCMSs and their organisations into layers. The allocation viewpoint defines how the executable components can be mapped into the physical devices. We validated our RA design via prototyping and surveying to measure the RA’s applicability in real-world scenarios and usability for stakeholders. Full article