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J. R. Ledesma, Meixin Zhang, Mei Fong Liew, R. Dominguez, Sophie M. Whikehart, M. Verma, Jianing Ma, Amanda Movo et al.

BACKGROUND Tuberculosis (TB) is the leading global cause of death from a single infectious agent. Recent reductions in global health funding have threatened TB control, making comprehensive assessment of TB, HIV-related TB, and drug-resistant TB burdens before these disruptions essential for shaping effective responses. The WHO End TB Strategy sets targets of a 95% reduction in TB deaths and a 90% reduction in TB incidence between 2015 and 2035. Using results from the Global Burden of Diseases, Injuries, and Risk Factors Study (GBD) 2023, this study aims to assess the burden of TB and multidrug-resistant TB (MDR-TB) across 204 countries and territories, and to evaluate progress towards the WHO End TB incidence and mortality targets. METHODS We quantified TB mortality using the Cause of Death Ensemble modelling platform with global vital registration, surveillance, verbal autopsy, and minimally invasive tissue sampling data. For TB morbidity estimation, we simultaneously modelled incidence, prevalence, and mortality by age and sex using DisMod-MR 2.1. A population attributable fraction (PAF) approach was applied to stratify morbidity and mortality estimates by HIV and drug-resistance status. We also calculated disability-adjusted life-years (DALYs) as the sum of years of life lost and years lived with disability. For the risk factor analysis, a comparative risk assessment framework was used and PAFs were derived for alcohol use, smoking, and high fasting plasma glucose to determine the proportion of TB burden associated with these risk factors. FINDINGS In 2023, there were an estimated 9·11 million (95% uncertainty interval 8·04-10·3) incident cases of all-form TB, 1·22 million (0·98-1·49) deaths, and 54·6 million (43·8-65·5) DALYs globally. HIV-related TB comprised 781 000 (690 000-879 000) incident cases and 210 000 (142 000-279 000) deaths, contributing 11·0 million (7·56-14·3) DALYs. MDR-TB accounted for 466 000 (198 000-1 080 000) incident cases, 102 000 (31 700-238 000) deaths, and 3·96 million (1·31-9·01) DALYs. From 2015 to 2023, global all-form TB incidence rates declined by 19·2% (17·8-20·5) and deaths declined by 22·6% (4·7-35·7); declines were larger for drug-susceptible TB than for MDR-TB. Sub-Saharan Africa and south Asia had the highest mortality burdens in 2023; reductions in all-form TB incidence and mortality were uneven between 2000 and 2023, with limited progress in both measures in Latin America and the Caribbean. Removing smoking, alcohol use, and high fasting plasma glucose would reduce global TB deaths to 768 000 (592 000-970 000) and DALYs to 34·9 million (27·8-43·8) in 2023; MDR-TB deaths would decrease to 77 200 (23 400-183 000) and DALYs to 3·12 million (1·03-7·29). INTERPRETATION Global progress towards WHO End TB targets is disparate and fragile. Although many regions achieved meaningful gains, others have stagnated in recent years. The complexity of TB prevention is amplified by divergent MDR-TB trends, the persistent burden of HIV, and growing exposure to modifiable risk factors. Recent volatility in global health financing threatens to further destabilise this vulnerable epidemiological landscape; concerted action is urgently needed to temper disruptions and preserve progress. FUNDING Gates Foundation.

Masoud Khazaei, Jann Harberts, A. Nilghaz, Michael Shola David, V. Cadarso, Muamer Dervisevic, N. Voelcker

Ensuring the quality, safety, and functional performance of stem cell cultures remains a critical challenge in biomedical research, drug development, and emerging cell-based therapies, as early metabolic disturbances can compromise outcomes and therapeutic efficacy. Glucose (Glu) consumption and L-lactate (LA) production serve as central indicators of cellular bioenergetic state, stress, and viability. However, conventional analytical approaches rely on discontinuous, labor-intensive assays that disrupt workflows and limit early intervention. Here, we report a miniaturized electrochemical biosensing platform for serial time-resolved analysis of Glu and LA in human induced pluripotent stem cell (hiPSC) culture media using a micropillar array (MPA)-based microfluidic electrochemical device (MED). The device exhibits linear detection ranges of 0.5-35 mM for Glu and 0.5-40 mM for LA, with limits of detection (LODs) of 0.18 ± 0.01 mM and 0.19 ± 0.01 mM, respectively. Using serially collected culture media, the MED quantifies dose-dependent metabolic responses to mitochondrial inhibition with oligomycin, revealing suppressed metabolic turnover and altered Glu-LA coupling prior to observable morphological changes. By enabling paired metabolic readouts from small-volume, undiluted culture-media samples, this platform provides a practical analytical route toward earlier recognition of culture-state deviations and more informed monitoring of stem-cell culture quality and drug-induced metabolic responses.

Avina Vongpradith, R. Dominguez, Lorainne Tudor Car, Amanda Movo, Samuel M. Ostroff, Jiawei He, Samuel B. Albertson, A. Carter et al.

BACKGROUND Enteric infectious diseases claim more than 1 million lives annually and are among the top ten causes of death in children younger than 5 years. Remarkable global investment has been dedicated to enteric infectious disease prevention and control; however, the shifting global health landscape is testing the continuance of progress. To evaluate the current status and guide future interventions, we present the latest epidemiological estimates of enteric infectious diseases from the Global Burden of Diseases, Injuries, and Risk Factors Study (GBD) 2023 and assess progress towards the Global Action Plan for the Prevention and Control of Pneumonia and Diarrhoea (GAPPD) mortality target of fewer than 20 deaths per 100 000 children younger than 5 years by 2025. METHODS We quantified the incidence, mortality, and disability-adjusted life-years (DALYs) of enteric infectious diseases by age, sex, and year across 204 countries and territories from 1990 to 2023. In GBD 2023, the following were considered under the category of enteric infectious diseases: diarrhoeal diseases, enteric fever (typhoid and paratyphoid), invasive non-typhoidal Salmonella spp (iNTS) infections, and other intestinal infectious diseases. We also examined 15 aetiologies contributing to diarrhoeal diseases. Incidence and prevalence were estimated with DisMod-MR (version 2.1), a Bayesian meta-regression tool, drawing on data from systematic reviews, population-based surveys, claims data, and hospital sources. Cause-specific mortality was modelled with Cause of Death Ensemble Modelling based on data from sources including vital registration, mortality surveillance, verbal autopsy, and minimally invasive tissue sampling. Years of life lost and years lived with disability were computed and combined to derive DALYs. For aetiology-specific estimation, population-attributable fractions (PAFs) for 15 pathogens were derived with a counterfactual framework. Point estimates and 95% uncertainty intervals (UIs) were generated from 250 draws from the posterior distribution. FINDINGS In 2023, enteric infectious diseases resulted in an estimated 1·27 million (95% UI 0·963-1·68) deaths globally, declining from 3·69 million (3·04-4·56) in 1990. The global age-standardised mortality rate (ASMR) decreased from 74·1 (62·0-92·9) per 100 000 population to 16·4 (12·6-21·3) per 100 000 population during the same period. Diarrhoeal diseases accounted for most deaths in 2023 (1·11 million [0·811-1·54]), followed by enteric fever and iNTS. South Asia and sub-Saharan Africa remained the most affected regions in 2023, with 599 000 (441 000-882 000) and 501 000 (373 000-648 000) deaths due to enteric infectious diseases, respectively, predominantly from diarrhoeal disease. Rotavirus was the leading cause of all-age diarrhoeal disease deaths (PAF 16·3% [12·0-21·5]), followed by norovirus (10·2% [2·4-17·0]) and Shigella spp (9·3% [5·4-15·2]). Among children younger than 5 years, PAFs of deaths due to diarrhoeal diseases were 40·2% (32·5-48·5) for rotavirus, 24·0% (15·1-36·7) for Shigella spp, and 23·4% (13·7-34·3) for adenovirus. Across 204 countries and territories, 141 met the GAPPD mortality target in 2023. The driving aetiologies among countries that did not meet the target in 2023 varied slightly by GBD super-region, but the highest or second-highest number of deaths in children younger than 5 years were consistently attributed to rotavirus. Astrovirus and sapovirus, newly included in GBD 2023, were responsible for 24 600 (6290-49 000) and 18 800 (4650-44 400) deaths, respectively, in 2023, mainly in children younger than 5 years. INTERPRETATION Our findings show that mortality and ASMRs of enteric infectious diseases declined substantially between 1990 and 2023. This decline is consistent with the expansion of public health measures and broader socioeconomic development. However, the burden in 2023 remains considerably high, with the highest mortality concentrated in sub-Saharan Africa and south Asia. Considering that more than a quarter of all countries had yet to meet the GAPPD mortality target in 2023, sustained efforts are needed to address the persistent burden in affected countries and to adapt to the changing global health landscape. FUNDING Gates Foundation.

Wildlife–vehicle collisions (WVCs) represent a growing safety, ecological, and economic challenge, with direct consequences for human lives, material damage, and biodiversity conservation. In the Federation of Bosnia and Herzegovina, systematic analyses that link traffic accidents caused by collisions with wildlife are lacking. This research identifies high-risk locations of WVCs and applies geospatial analysis to the main roads of the Federation of Bosnia and Herzegovina. The analysis is based on official police reports documenting 14,169 traffic accidents between 2021 and 2023, of which 104 cases (0.73%) were classified as animal-related. Although species were not specified in the reports, these accidents predominantly occurred in areas where wildlife crossings are expected, and thus are treated as potential wildlife–vehicle collisions. The results indicate a concentration of WVCs in nine municipalities, with eight critical road segments identified on main roads. Additional analyses explored the relationship between collisions, road infrastructure (bridges, tunnels), and ecological features of habitats (Emerald Network, Natura 2000, Red List of FBiH, IUCN). Based on the findings, it can be concluded that spatially targeted prevention is essential, with priority given to infrastructural measures (wildlife overpasses, fencing, signage) and strategic measures (improved databases, continuous monitoring, and integration into spatial planning). The obtained results provide a foundation for policies that simultaneously enhance traffic safety and contribute to the protection of wildlife populations.

Sarah Brooke Sirota, Rose G. Bender, R. Dominguez, Avina Vongpradith, Amanda Movo, Lucien R. Swetschinski, Daniel T Araki, Chieh Han et al.

BACKGROUND Meningitis remains the leading infectious cause of neurological disabilities globally, disproportionately affecting children younger than 5 years and populations in the African meningitis belt. Whereas previous global estimates focused on ten pathogen categories, this study presents the most comprehensive analysis to date, assessing the meningitis burden attributable to 17 causative pathogens based on the Global Burden of Diseases, Injuries, and Risk Factors Study (GBD) 2023 framework. METHODS GBD is a systematic, scientific effort aimed at quantifying the comparative magnitude of health loss caused by diseases, injuries, and risk factors across age groups, sexes, and geographical locations over time. We estimated meningitis mortality using the Cause of Death Ensemble model (CODEm) and morbidity using DisMod-MR 2.1, incorporating data from vital registration, verbal autopsy, surveillance, hospital data, and systematic reviews. Aetiology-specific estimates were generated with pathogen-linked case-fatality ratios and splined binomial regression models. Risk factor attribution was based on established risk-outcome pairs and population attributable fractions. FINDINGS In 2023, there were 259 000 (95% uncertainty interval 202 000-335 000) global deaths and 2·54 million (2·20-2·93) incident cases of meningitis. Children younger than 5 years accounted for more than a third of deaths (86 600 [53 300-149 000]). Streptococcus pneumoniae, Neisseria meningitidis, non-polio enteroviruses, and other viruses were the leading causes of death, while non-polio enteroviruses caused the most cases. The four WHO-defined preventable meningitis pathogens of interest (S pneumoniae, N meningitidis, Haemophilus influenzae, and Group B streptococcus) contributed to 98 700 deaths (77 000-127 000) and 594 000 cases (514 000-686 000). Low birthweight, short gestation, and household air pollution were the top risk factors for meningitis-related mortality. INTERPRETATION Although mortality and incidence have declined significantly since 1990, progress is insufficient to meet WHO 2030 targets. Despite marked progress in reducing bacterial meningitis via global vaccination campaigns, a substantial meningitis burden persists, attributable both to common pathogens such as S pneumoniae and N meningitidis and to emerging non-bacterial pathogens such as Candida spp and drug-resistant fungi. Achieving WHO goals will require sustained investment in surveillance, vaccination, maternal screening, and health-system strengthening, especially in high-burden settings. FUNDING Gates Foundation, Wellcome Trust, and UK Department of Health and Social Care.

K. Bhangdia, Miranda L May, Jonathan M Kocarnik, Natalie Pritchett, Andrew Crist, Louise Penberthy, Alistair Acheson, Lee Deitesfeld et al.

BACKGROUND Breast cancer is a leading cause of mortality and morbidity among females worldwide. As part of the Global Burden of Diseases, Injuries, and Risk Factors Study (GBD) 2023, we provided an updated comprehensive assessment of the epidemiological trends, disease burden, and risk factors associated with breast cancer globally, regionally, and nationally from 1990 to 2023. METHODS Breast cancer incidence, mortality, prevalence, years lived with disability (YLDs), years of life lost (YLLs), and disability-adjusted life-years (DALYs) were estimated by age and sex for 204 countries and territories from 1990 to 2023. Mortality estimates were generated using GBD Cause of Death Ensemble models, leveraging data from population-based cancer registration systems, vital registration systems, and verbal autopsies. Mortality-to-incidence ratios were calculated to derive both mortality and incidence estimates. Prevalence was calculated by combining incidence and modelled survival estimates. YLLs were established by multiplying age-specific deaths with the GBD standard life expectancy at the age of death. YLDs were estimated by applying disability weights to prevalence estimates. The sum of YLLs and YLDs equalled the number of DALYs. Breast cancer burden attributable to seven risk factors was examined through the comparative risk assessment framework. The GBD forecasting framework was used to forecast breast cancer incidence and mortality from 2024 to 2050. Age-standardised rates were calculated for each metric using the GBD 2023 world standard population. FINDINGS In 2023, there were an estimated 2·30 million (95% uncertainty interval [UI] 2·01 to 2·61) breast cancer incident cases, 764 000 deaths (672 000 to 854 000), and 24·1 million (21·3 to 27·5) DALYs among females globally. In the World Bank low-income group, where a low age-standardised incidence rate (ASIR) was estimated (44·2 per 100 000 person-years [31·2 to 58·4]), the age-standardised mortality rate (ASMR) was the highest (24·1 per 100 000 [16·8 to 31·9]). The highest ASIR was in the high-income group (75·7 per 100 000 [67·1 to 84·0]), and the lowest ASMR was in the upper-middle-income group (11·2 per 100 000 [10·2 to 12·3]). Between 1990 and 2023, the ASIR in the low-income group increased by 147·2% (38·1 to 271·7), compared with a 1·2% (-11·5 to 17·2) change in the high-income group. The ASMR decreased in the high-income group, changing by -29·9% (-33·6 to -25·9), but increased by 99·3% (12·5 to 202·9) in the low-income group. The increase in age-standardised DALY rates followed that of ASMRs. Risk factors such as dietary risks, tobacco use, and high fasting plasma glucose contributed to 28·3% (16·6 to 38·9) of breast cancer DALYs in 2023. The risk factors with a decrease in attributable DALYs between 1990 and 2023 were high alcohol use and tobacco. By 2050, the global incident cases of breast cancer among females were forecast to reach 3·56 million (2·29 to 4·83), with 1·37 million (0·841 to 2·02) deaths. INTERPRETATION The stable incidence and declining mortality rates of female breast cancer in high-income nations reflect success in screening, diagnosis, and treatment. In contrast, the concurrent rise in incidence and mortality in other regions signals health system deficits. Without effective interventions, many countries will fall short of the WHO Global Breast Cancer Initiative's ambitious target of achieving an annual reduction of 2·5% in age-standardised mortality rates by 2040. The mounting breast cancer burden, disproportionately affecting some of the world's most vulnerable populations, will further exacerbate health inequalities across the globe without decisive immediate action. FUNDING Gates Foundation, St Jude Children's Research Hospital.

Foroogh Rezaei, Siu Wai Wong, Muamer Dervisevic, Xenia Kostoulias, Yue Qu, Beatriz Prieto-Simón, J. Baell, N. Voelcker et al.

Rapid detection of antibiotic-resistant bacteria is a crucial tool in the global fight against antimicrobial resistance, helping to limit the spread of resistance and guide treatment decisions. Here, we report the design, synthesis, and electrochemical evaluation of β-lactam-based redox-activatable probes for detecting β-lactamase activity. The probes incorporate a β-lactam core linked to redox reporters through cleavable linkages, enabling signal generation upon enzymatic hydrolysis. High-performance liquid chromatography and differential pulse voltammetry analyses were used to assess time-dependent activation and concentration-dependent responses against commercial β-lactamase blends and metallo-β-lactamases. Selected probes, bearing cephalosporin recognition motifs and maltol redox reporters, were further evaluated against clinical isolates, demonstrating selective activation in carbapenemase-producing strains. To extend the platform toward solid-state biosensing, an azide-functionalized analog was clicked on alkyne-modified glassy carbon electrodes. Stepwise surface functionalization and immobilization were validated electrochemically using model redox reporters, confirming their activity. The immobilized probe retained responsiveness, demonstrating the feasibility of integrating this sensing strategy into solid-state diagnostic devices. By integrating stable cephalosporin scaffolds with redox-reporter signaling, this work introduces a novel probe system that unites chemical probe design with surface-based electrochemical sensing, providing a strong foundation for the development of portable, point-of-care diagnostics for β-lactamase-mediated antibiotic resistance.

Masoud Khazaei, Seyedeh Somayeh Hosseinikebria, Muamer Dervisevic, Jingliang Li, J. Razal, N. Voelcker, Azadeh Nilghaz

Food, especially fish meat, is extremely vulnerable to oxidation and microbiological deterioration. Therefore, effective analytical techniques for quality control and safety monitoring are required. Electrochemical biosensors have become reliable, rapid, and affordable devices for in-field and real-time food quality assessment. However, their application is often limited in point-of-need scenarios due to the requirement for intensive sample preparation. Here, we introduce a microneedle array (MNA)-based electrochemical biosensor, designed for direct food safety and quality analysis without the need for sample preparation. A gold (Au)-coated polymeric MNA was functionalized with a chitosan-gold nanoparticles (Ch-AuNP) nanocomposite and further modified by immobilizing xanthine oxidase (XO) for selective hypoxanthine (HX) detection. The MNA-based biosensor exhibited a linear range between 5 and 50 μM, and 50 to 200 μM, with a sensitivity of 0.024 μA/μM and a limit of detection (LOD) of 2.18 ± 0.75 μM for HX, with a response time of approximately 100 s. Furthermore, the MNA-based biosensor was successfully utilized for monitoring HX levels in fish tissue samples over 48 h, showing strong agreement with results obtained from a commercial Amplex Red assay kit. The technology can be used for real-time food quality assessment and food safety monitoring due to its high sensitivity, interference tolerance, and lack of requirement for sample preparation.

Sarah Brooke Sirota, Rose G. Bender, R. Dominguez, Amanda Movo, Lucien R. Swetschinski, Daniel T Araki, Chieh Han, Eve E Wool et al.

BACKGROUND Lower respiratory infections (LRIs) remain the world's leading infectious cause of death. This analysis from the Global Burden of Diseases, Injuries, and Risk Factors Study (GBD) 2023 provides global, regional, and national estimates of LRI incidence, mortality, and disability-adjusted life-years (DALYs), with attribution to 26 pathogens, including 11 newly modelled pathogens, across 204 countries and territories from 1990 to 2023. With new data and revised modelling techniques, these estimates serve as an update and expansion to GBD 2021. Through these estimates, we also aimed to assess progress towards the 2025 Global Action Plan for the Prevention and Control of Pneumonia and Diarrhoea (GAPPD) target for pneumonia mortality in children younger than 5 years. METHODS Mortality from LRIs, defined as physician-diagnosed pneumonia or bronchiolitis, was estimated using the Cause of Death Ensemble model with data from vital registration, verbal autopsy, surveillance, and minimally invasive tissue sampling. The Bayesian meta-regression tool DisMod-MR 2.1 was used to model overall morbidity due to LRIs. DALYs were calculated as the sum of years of life lost (YLLs) and years lived with disability (YLDs) for all locations, years, age groups, and sexes. We modelled pathogen-specific case-fatality ratios (CFRs) for each age group and location using splined binomial regression to create internally consistent estimates of incidence and mortality proportions attributable to viral, fungal, parasitic, and bacterial pathogens. Progress was assessed towards the GAPPD target of less than three deaths from pneumonia per 1000 livebirths, which is roughly equivalent to a mortality rate of less than 60 deaths per 100 000 children younger than 5 years. FINDINGS In 2023, LRIs were responsible for 2·50 million (95% uncertainty interval [UI] 2·24-2·81) deaths and 98·7 million (87·7-112) DALYs, with children younger than 5 years and adults aged 70 years and older carrying the highest burden. LRI mortality in children younger than 5 years fell by 33·4% (10·4-47·4) since 2010, with a global mortality rate of 94·8 (75·6-116·4) per 100 000 person-years in 2023. Among adults aged 70 years and older, the burden remained substantial with only marginal declines since 2010. A mortality rate of less than 60 deaths per 100 000 for children younger than 5 years was met by 129 of the 204 modelled countries in 2023. At a super-regional level, sub-Saharan Africa had an aggregate mortality rate in children younger than 5 years (hereafter referred to as under-5 mortality rate) furthest from the GAPPD target. Streptococcus pneumoniae continued to account for the largest number of LRI deaths globally (634 000 [95% UI 565 000-721 000] deaths or 25·3% [24·5-26·1] of all LRI deaths), followed by Staphylococcus aureus (271 000 [243 000-298 000] deaths or 10·9% [10·3-11·3]), and Klebsiella pneumoniae (228 000 [204 000-261 000] deaths or 9·1% [8·8-9·5]). Among pathogens newly modelled in this study, non-tuberculous mycobacteria (responsible for 177 000 [95% UI 155 000-201 000] deaths) and Aspergillus spp (responsible for 67 800 [59 900-75 900] deaths) emerged as important contributors. Altogether, the 11 newly modelled pathogens accounted for approximately 22% of LRI deaths. INTERPRETATION This comprehensive analysis underscores both the gains achieved through vaccination and the challenges that remain in controlling the LRI burden globally. Furthermore, it demonstrates persistent disparities in disease burden, with the highest mortality rates concentrated in countries in sub-Saharan Africa. Globally, as well as in these high-burden locations, the under-5 LRI mortality rate remains well above the GAPPD target. Progress towards this target requires equitable access to vaccines and preventive therapies-including newer interventions such as respiratory syncytial virus monoclonal antibodies-and health systems capable of early diagnosis and treatment. Expanding surveillance of emerging pathogens, strengthening adult immunisation programmes, and combating vaccine hesitancy are also crucial. As the global population ages, the dual challenge of sustaining gains in child survival while addressing the rising vulnerability in older adults will shape future pneumonia control strategies. FUNDING Gates Foundation.

Yuexi Lin, Muniba Bhatti, Michael Shola David, Yannic Brasse, Jann Harberts, Thomas Kister, Muamer Dervisevic, Tobias Kraus et al.

Wearable electrochemical biosensors offer a promising alternative to conventional invasive blood‐based methods for monitoring biomarkers in diagnostic or therapeutic applications. Microneedle (MN)‐based technology provides direct access to the skin's interstitial fluid (ISF), enabling real‐time monitoring of biomarkers. Nevertheless, current micro‐ and nanofabrication techniques do not adequately support the development of MN‐based wearable technology that can utilize soft hybrid conductive inks, limiting its use in transdermal biosensing. Herein, an MN‐based biosensing platform is developed by integrating 3D printing, soft lithography, and hybrid conductive ink technology, featuring a fenestrated MN shell (FMNS) that serves as a protective layer for the inner hybrid conductive ink coating and prevents delamination during skin application. This FMNS patch demonstrates a wide pH monitoring range, high selectivity and accurate detection of subtle ISF pH changes, safe integration of hybrid conductive inks, and reduced fabrication time and cost when compared to other microfabrication methods such as lithography and deep reactive ion etching. The biosensor excels in protecting the biosensing layer and demonstrates excellent analytical performance in monitoring changes in pH levels of the skin ISF. This micro‐ and nanofabrication approach has great potential in integrating hybrid conductive ink technology into transdermal wearable devices for health monitoring and diagnostics.

Michael Shola David, Jann Harberts, Raquel Sanchez Salcedo, Muamer Dervisevic, V. Cadarso, N. Voelcker

Mammalian cells, particularly human cell culture models, are essential for studying disease pathophysiology and producing cell-based therapeutic products. Monitoring and controlling cell culture conditions accurately is essential for optimal cell growth and health, as even minor variations can significantly influence cell behavior. The presence of viruses, bacteria, and their by-products are key indicators of cell culture contamination. Conventional assays for quantifying cellular health and microbial contaminants such as endotoxins are end-point assays that are often laborious, require specialized equipment, and typically detect contamination only at advanced stages. For example, the chromogenic Limulus amoebocyte lysate assay, used for quantifying endotoxin, a bacte rial by-product, is often susceptible to interference from serum proteins in the culture medium. In this work, we present a simple and sensitive aptamer-based biosensor designed to detect bacterial-secreted endotoxins in various complex cell culture media. As a proof of concept, human induced pluripotent stem cells (hiPSCs) were deliberately contaminated with Escherichia coli (E. coli), and the biosensor's response to endotoxins released by the bacteria was monitored over a 24-h period. The biosensor demonstrated a reliable linear response with a detection limit of 0.33 ± 0.06 pg/mL in DMEM and 0.142 ± 0.025 pg/mL in StemFlex medium. Its performance in complex sample matrices suggests the potential for integration with industrial-scale cell culture systems for real-time contamination detection, providing a cost-effective, efficient, and timely method to monitor cell health and ensure sterile conditions for therapeutic cell cultivation.

Masoud Khazaei, Jann Harberts, Azadeh Nilghaz, M. David, K. Galbraith, Muamer Dervisevic, V. Cadarso, N. Voelcker

Glucose levels serve as a fundamental indicator of cell health, reflecting crucial aspects of cellular metabolism and energy production. While effective, traditional methods such as spectrophotometry and chromatography have limitations, such as labour-intensive sample collection, reliance on bulky equipment, extensive sample preparation, and prolonged experimental durations. To address these issues, we introduce a micropillar-based microfluidic electrochemical device (MED) for real-time monitoring of glucose levels in diverse cell culture systems, including human induced pluripotent stem cells (hiPSCs) and murine fibroblast cells (GP + E86). This biosensor demonstrates a linear range of 0.025-1.50 mM and a high sensitivity of 4.71 ± 0.13 μA. mM-1, and a low limit of detection of 19.10 ± 0.50 μM. The MED not only delivered fast glucose measurements with accuracy and reliability comparable to ultra-high-performance liquid chromatography (UHPLC) but was also specifically evaluated on GP + E86 murine fibroblast cells at varying seeding densities (1:5 and 1:10 ratios), across different culturing times to accurately monitor dynamic metabolic shifts associated with various growth phases. Furthermore, the MED effectively detected significant changes in glucose consumption in hiPSCs cell cultures contaminated with Escherichia coli (E. coli), highlighting its potential for early contamination detection. Integrating non-invasive, continuous monitoring platforms enhances the reliability of experimental outcomes by enabling cell health monitoring without disrupting the cell culture process. This approach enables real-time monitoring of cell cultures ensuring accurate detection of metabolic changes and early detection of media contamination.

Yuexi Lin, Muamer Dervisevic, H. Yoh, Keying Guo, N. Voelcker

Microneedles (MNs) are emerging as versatile tools for both therapeutic drug delivery and diagnostic monitoring. Unlike hypodermic needles, MNs achieve these applications with minimal or no pain and customizable designs, making them suitable for personalized medicine. Understanding the key design parameters and the challenges during contact with biofluids is crucial to optimizing their use across applications. This review summarizes the current fabrication techniques and design considerations tailored to meet the distinct requirements for drug delivery and biosensing applications. We further underscore the current state of theranostic MNs that integrate drug delivery and biosensing and propose future directions for advancing MNs toward clinical use.

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