Abstract Airborne micro- and nanoplastics are emerging inhalation contaminants, yet their effects on the first human airway barrier, the nasal epithelium, remain poorly understood. Here, we investigated metabolic responses of normal human nasal epithelial cells exposed to polystyrene nanoplastics (100 nm), microplastics (7 μm), or their mixture using comprehensive metabolomics and lipidomics of both cells and secretome analyzed by ultrahigh-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry. Nanoplastics induced the strongest intracellular perturbations, characterized by increased amino acid- and purine-related metabolites, including homocystine, uric acid, aspartic acid, and glutamic acid, together with marked depletion of membrane lipids, particularly phosphatidylcholines. The particle mixture produced partially overlapping but generally attenuated responses, whereas microplastics alone caused comparatively modest changes. This attenuation was consistent with particle characterization showing partial nanoplastic adsorption onto microplastics, which reduced the freely dispersed nanoplastic fraction in the mixture. Secretome profiling revealed distinct extracellular metabolic fingerprints, with nanoplastics and mixtures increasing metabolites linked to oxidative stress, amino acid turnover, and detoxification, while reducing several membrane-derived lipids. Pathway enrichment analysis highlighted disturbances in carbohydrate metabolism, glycerophospholipid metabolism, the pentose phosphate pathway, and amino acid and nucleotide metabolism. Overall, these findings identify normal human nasal epithelial cells as a metabolically sensitive target of plastic particle exposure and suggest that nanoplastics are primary drivers of the observed metabolic perturbations at the airway interface in vitro.

Titanium dioxide nanoparticles (TiO₂NPs) are widely produced engineered nanomaterials with ongoing human exposure through consumer and occupational uses. Conventional in vitro assays often focus on cytotoxicity and may therefore overlook early or sublethal cellular perturbations. Here, we applied Cell Painting-based phenomics to resolve size-dependent sub-lethal phenotypic signatures of TiO2NP exposure in human HepG2 hepatocytes. Two TiO2NPs (<25 nm and <100 nm) were characterized by field emission scanning electron microscopy and evaluated following 24-hour exposure at five concentrations: 6.25, 12.5, 25, 50, and 100 µg/mL. Cell viability was assessed using the alamarBlue assay, and high-dimensional phenotypic profiles were generated using Cell Painting-based phenomics, including automated high-content imaging and CellProfiler-based feature extraction. TiO2NP exposure induced modest reductions in viability at the highest concentration, indicating limited acute cytotoxicity. In contrast, phenomic profiling revealed clear, concentration-dependent phenotypic perturbations for both size fractions, with markedly stronger and more consistent effects for the < 100 nm TiO2NPs. At 100 µg/mL, the < 100 nm TiO2NPs altered 50.9% of the measured phenotypic features, compared with 28.9% for the < 25 nm particles, with prominent contributions from endoplasmic reticulum-, actin/Golgi/plasma membrane-, mitochondria-, and RNA-associated features. Dimensionality reduction and correlation analyses confirmed reproducible, concentration-dependent phenotypic trajectories. Importantly, the TiO2NP-induced phenotypes were distinct from those induced by the reference chemical CA-074Me, which produced broad perturbations and served as a reference chemical to verify assay sensitivity and dynamic range. Overall, Cell Painting phenomics sensitively captures size-dependent, sublethal cellular phenotypes induced by TiO2NPs, supporting its value as a New Approach Methodology for nanosafety assessment beyond conventional viability endpoints.

The aim of the study was to determine nano/sub-micron particle and dust exposure levels throughout the whole workflow at a Swedish metal additive manufacturing (AM) facility, focusing on the laser powder bed fusion (L-PBF) method. By evaluating particle levels and composition across different AM processes using both stationary and personal sampling, the study sought to improve exposure assessment and inform protective measures in the metal AM workplaces. Measurements were conducted during five measurement weeks, as five working days Monday-Friday, between October 2020 and October 2023. Personal particle measurements in the breathing zone were performed on Mondays and Fridays for 1 to 3 workers per day. Stationary particle and dust sampling were performed continuously at three locations each week to capture task-specific and temporal variation in emissions. Nano/sub-micron particle concentrations ranged from 0 to 3.3 million particles/cm(3), with the highest peaks recorded in the post-processing area. Elevated levels were also detected, near the depowdering machine, by the bandsaw, and in the lunchroom, while levels near the printers were low (<10,000 particles/cm(3)). Personal exposure peaks occurred during printer cleaning, feedstock powder filling, dust removal with compressed air, post-processing, and packing. In contrast to the increased nano/sub-micron particle levels observed, respirable, and inhalable dust levels were very low. The study highlights the need to monitor particle exposure during both manufacturing and post-processing. Health risks associated with airborne particles are influenced by both exposure levels and the toxicological properties of materials. To ensure a safe and sustainable future for metal AM, comprehensive exposure assessment, risk evaluation, and the implementation of protective measures remain essential.

Nanoplastics have recently been detected in human liver tissue, raising concerns about their potential impact on liver function. However, early hepatocyte responses associated with nanoplastics exposure remain poorly understood. Here, we combined high-throughput Cell Painting-based phenomics, untargeted metabolomics, and Seahorse mitochondrial functional assay to investigate the effects of 100 nm polystyrene nanoplastics on human HepaRG hepatocytes, a surrogate for primary human hepatocytes. At the tested concentrations (6.25-100 µg/mL), exposure did not induce overt cytotoxicity, enabling assessment of early sublethal cellular responses. Phenomics revealed widespread subcellular perturbations, with 16.4% of the measured phenotypic features significantly altered. Mitochondria-associated features represented the dominant altered phenotypic signature, showing pronounced changes in granularity, texture, and radial distribution, alongside alterations in endoplasmic reticulum- and cytoskeleton-associated features. Untargeted metabolomics of intracellular metabolites and the extracellular secretome revealed metabolic alterations, characterized by changes consistent with altered β-oxidation, lipid handling, membrane stress, and central carbon metabolism, including changes in the tricarboxylic acid (TCA) cycle and amino acid catabolism. Pathway analysis identified the TCA cycle as one of the most significantly affected pathways (FDR = 0.028). Integrated phenomic-metabolomic analysis revealed strong correlations between mitochondrial phenotypic features and metabolites involved in lipid and energy metabolism, indicating a coordinated structural-metabolic response to polystyrene nanoplastics exposure. Functional assessment using Seahorse assay showed reduced basal and maximal respiration and decreased ATP-linked O2 consumption. Together, these findings provide evidence that 100 nm polystyrene nanoplastics elicit early mitochondria-associated phenotypic, metabolic, and functional responses prior to overt cytotoxicity under the tested conditions. They also highlight the value of phenomic-metabolomic-functional integration for profiling sublethal nanotoxicological responses and guiding future targeted mechanistic studies.

Abstract Background Respirable crystalline silica (RCS) is a well-established occupational health hazard. Inflammation and oxidative stress are key mechanisms for diseases associated with exposure; however, the role of lipid inflammatory mediators remains unclear. Oxylipins, a diverse group of lipid mediators derived mainly from arachidonic acid or linoleic acid via cyclooxygenases, lipoxygenases or P450 enzymes may contribute to silica-induced pathology. This study investigates the association between RCS exposure and oxylipin/oxidative stress biomarkers in a cohort of foundry workers. Method Plasma levels of 79 lipid mediators, including oxylipins and un- mono- and polyunsaturated fatty acids were quantified in 40 foundry workers sampled twice, using solid-phase extraction and ultra high-pressure liquid chromatography coupled to unispray tandem mass spectrometry (SPE-UHPLC-USI-MS/MS). Lipid levels were correlated with personal exposure measurements to respirable dust and RCS. Analysis of urinary oxylipin and oxidative stress markers are ongoing and these results will also be presented at the conference. Results Adjusted for covariates, seven lipids were significantly correlated with RCS exposure. These lipids were mainly linoleic acid derived oxylipins with 9,10-DiHOME demonstrating the strongest correlation. For respirable dust, five lipids correlated significantly with exposure, mainly eicosanoids, eg 8,9-DiHETrE. In general, there were no, or very few significant correlations between the lipid mediators and other inflammatory markers, such as interleukin-8, C-reactive protein and Serum amyloid A. Conclusions The observed associations between linoleic acid-derived oxylipins and RCS exposure suggest that these lipid mediators may play a previously underrecognized role in silica-induced inflammation and disease.

BACKGROUND & AIMS:To investigate the role of surgical technique - OPEN or minimally invasive surgery (MIS) - on the development of postoperative insulin resistance and inflammatory response in elective colorectal resections for cancer. METHODS:Clinical exploratory study of patients with colorectal cancer planned for elective colorectal resections by either OPEN (n = 8) or MIS (n = 9) approach within a controlled Enhanced Recovery After Surgery (ERAS) program. Insulin sensitivity was determined using the hyperinsulinemic normoglycemic clamp and blood samples were collected before and after surgery for analysis of glucose, hormones and inflammatory markers in both the OPEN and MIS group, as well as comparing the two groups to each other. RESULTS:Patients undergoing surgery, OPEN or MIS, exhibited postoperative insulin resistance to a similar degree regardless of surgical approach. Postoperative plasma levels of the inflammatory markers CRP and IL-6 were significantly (p < 0.05) higher following open surgery. Perioperative compliance to the ERAS protocol was similar in both the OPEN and MIS groups. CONCLUSION:While open colorectal surgery did elicit a significantly higher inflammatory response compared to MIS, the surgical approach did not influence the degree of postoperative insulin resistance in patients undergoing colorectal resections for cancer in ERAS.

Meningococcal disease is caused by the bacterium Neisseria meningitidis, which is a commensal in the human upper airways. Teenagers and young adults are the main reservoir for the bacteria. The aim of this study was to assess the prevalence of meningococcal carriage and risk factors for carriage among high school and university students in Northwestern Ethiopia. Oropharyngeal swabs and buccal samples were collected and stored on Whatman FTA-cards pending PCR analysis. DNA was extracted and a Multiplex TaqMan® custom assay was performed to detect N. meningitidis bacteria using the genes sodC and porA as species-specific targets. Genogrouping was performed for the samples positive for N. meningitidis using multiplex real-time PCR for the groups A, B, C, W, Y and X using a Rotor-Gene Q real-time PCR system. Out of the 1025 participants, 52

The host immune response in sepsis involves both pro- and anti-inflammatory mechanisms, with monocytes playing a central role in the process. We have previously identified an in vitro response profile of endotoxin (LPS) tolerant primary human monocytes, consisting of eight cytokines/chemokines as well as a set of five transcription factors. In the current study, we evaluated differences in expression levels of these investigated molecular markers across different patient groups (patients with or without infection, and with or without sepsis), and their association with clinical outcomes (septic shock and in-hospital mortality), among 809 ambulance patients. The results showed that patients with sepsis displayed the lowest HLA-DRA expression levels together with the lowest TNF/IL-10 ratio, while most other cytokine/chemokines and gene expressions were elevated. Higher levels of HGF, CCL8, CCL2, TNF and IL-10, as well as upregulation of HIF1A and NFKBIA were seen in septic patients with septic shock. The data suggests that the investigated immunological markers linked to immunosuppressed monocyte responses are associated with patients with sepsis and septic shock.

Metal additive manufacturing (AM) relies on alloy feedstock powders that may come into contact with the workers' skin during handling, yet skin-relevant data on metal release and biological reactivity remain limited. Here, we assessed the cutaneous bioactivity of the fine particle fraction of four gas-atomized Fe-based AM powders (316L stainless steel, Fe-powder A, and tooling steels B and C). Powders were sieved to <10 μm and characterized by scanning electron microscopy and X-ray photoelectron spectroscopy before and after incubation in artificial sweat (ASW). Metal biodissolution was quantified in ASW and keratinocyte culture medium using atomic absorption spectrophotometry. Cellular responses were evaluated in HaCaT keratinocytes using Cell Painting-based phenomics and multiplex cytokine/chemokine profiling and in an ex vivo full-thickness human skin explant model, including superficial barrier disruption, IL-8/CXCL8 quantification, and histological assessment. ASW exposure induced marked shifts in the outermost surface composition across powders, indicating sweat-driven surface transformation. Biodissolution was low and medium-dependent, with Fe dominating the release in ASW, and with an overall metal release remaining limited in cell culture medium. In HaCaT cells, MCP-1/CCL2, IL-6, and IL-8/CXCL8 were quantifiable but showed no significant changes following powder exposure. Cell Painting revealed subtle, shared phenotypic signatures, primarily involving mitochondrial-associated features, without evidence of broad cellular stress. In the ex vivo skin model, AM powders did not increase IL-8/CXCL8 secretion, the particles remained localized to the skin surface without detectable penetration, and coexposure with Staphylococcus epidermidis did not enhance bacterial colonization or induce inflammation. To the best of our knowledge, this is the first study that applies a human skin explant model to evaluate dermal responses to metal AM powders. Overall, the tested AM powders showed low short-term cutaneous reactivity under skin-relevant conditions, providing human-relevant evidence to inform occupational risk assessment in AM environments.

Per- and polyfluoroalkyl substances and nanoplastics frequently co-occur in environmental matrices, yet the effects of co-exposure on cellular responses upon ingestion are poorly understood. Here, we exposed human intestinal Caco-2 cells to perfluorooctanesulfonic acid, nanoplastics, and their combination. Cell painting-based phenomics was used to map phenotypic alterations across subcellular structures, and untargeted metabolomics using ultra-high-performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry was employed to assess metabolic changes. Results show that perfluorooctanesulfonic acid predominantly affected the actin cytoskeleton, Golgi apparatus, and plasma membrane, while nanoplastics primarily targeted mitochondria. Combined exposure disrupted the endoplasmic reticulum, RNA, and mitochondria. Perfluorooctanesulfonic acid reduced levels of carnitines, free fatty acids, nucleotides, and sugars, whereas nanoplastics inhibited ceramides, triglycerides, sphingomyelins, and additional free fatty acids. Combined exposure produced a metabolic profile resembling that of nanoplastics, with specific differences attributed to perfluorooctanesulfonic acid. Overall, nanoplastics appear as the main drivers of the co-exposure effects.

As the volume of plastic waste from electrical and electronic equipment (WEEE) continues to rise, a significant portion is disposed of in the environment, with only a small fraction being recycled. Both disposal and recycling pose unknown health risks that require immediate attention. Existing knowledge of WEEE plastic toxicity is limited and mostly relies on epidemiological data and association studies, with few insights into the underlying toxicity mechanisms. Therefore, this study aimed to perform comprehensive chemical screening and mechanistic toxicological assessment of WEEE plastic-associated chemicals. Chemical analysis, utilizing suspect screening based on high-resolution mass spectrometry, along with quantitative target chemical analysis, unveiled numerous hazardous compounds including polyaromatic compounds, organophosphate flame retardants, phthalates, benzotriazoles, etc. Toxicity endpoints included perturbation of morphological phenotypes using the Cell Painting assay, inflammatory response, oxidative stress, and endocrine disruption. Results demonstrated that WEEE plastic chemicals altered the phenotypes of the cytoskeleton, endoplasmic reticulum, and mitochondria in a dose-dependent manner. In addition, WEEE chemicals induced inflammatory responses in resting macrophages and altered inflammatory responses in lipopolysaccharide-primed macrophages. Furthermore, WEEE chemicals activated the nuclear factor erythroid 2-related factor 2 (Nrf2) pathway, indicating oxidative stress, and the aryl hydrocarbon receptor (AhR). Endocrine disruption was also observed through the activation of estrogenic receptor-α (ER-α) and the induction of anti-androgenic activity. The findings show that WEEE plastic-associated chemicals exert effects in multiple subcellular sites, via different receptors and mechanisms. Thus, an integrated approach employing both chemical and toxicological methods is essential for comprehensive assessment of the toxicity mechanisms and cumulative chemical burden of WEEE plastic-associated chemicals.

The environmental fragmentation of plastics generates a mixture of plastic particles of various sizes, which frequently co-occur with other mobile and persistent environmental pollutants. Despite the prevalence of such scenarios, the interaction between micro- and nanoplastics (MNPs) and their combined effects with environmental pollutants, such as highly toxic hexavalent chromium (Cr(VI)), remain almost entirely unexplored in mammalian species. This study demonstrated that nanoplastic and microplastic particles co-aggregate and together influence Cr bioaccumulation patterns and related physiological alterations in rats. Following a four-week repeated intragastric exposure of Wistar rats to MNPs and Cr(VI), either alone or in combination, MNPs significantly enhanced Cr bioaccumulation in the liver, heart, brain, and skin. Under co-exposure conditions, Cr(VI) was the primary driver of cellular effects observed in the blood, including shifts in immune cell subpopulations (e.g., neutrophils, lymphocytes) and alterations in red blood cell indices, while serum biochemistry reflected limited physiological stress. MNPs per se decreased creatine kinase activity and increased cholesterol levels. In summary, polystyrene MNPs increase Cr(VI) distribution and bioavailability, but co-exposure does not uniformly exacerbate toxicity. Instead, their interaction may selectively alter physiological responses, emphasizing the need for a deeper understanding of their combined effects and potential health risks.

Existing research has proven difficult to understand the interplay between upstream signaling events during NLRP3 inflammasome activation. Additionally, events downstream of inflammasome complex formation such as cytokine release and pyroptosis can exhibit variation, further complicating matters. Cell Painting has emerged as a prominent tool for unbiased evaluation of the effect of perturbations on cell morphological phenotypes. Using this technique, phenotypic fingerprints can be generated that reveal connections between phenotypes and possible modes of action. To the best of our knowledge, this was the first study that utilized Cell Painting on human THP-1 macrophages to generate phenotypic fingerprints in response to different endogenous and exogenous NLRP3 inflammasome triggers and to identify phenotypic features specific to NLRP3 inflammasome complex formation. Our results demonstrated that not only can Cell Painting generate morphological fingerprints that are NLRP3 trigger-specific but it can also identify cellular fingerprints associated with NLRP3 inflammasome activation.

The rapid rise of 3D printing, both in industrial and home settings, presents emerging health and environmental risks. While 3D printing enhances sustainability by reducing waste and optimizing resource use, its impact on human health remains poorly understood. The use of metals and polymers linked to health risks, coupled with the release of inhalable particles and volatile organic compounds, raises concerns about respiratory and systemic effects. The absence of clear guidelines creates high public demand for information and limits safe implementation, particularly in schools and homes where millions of 3D printers are expected by 2030. Additionally, improper disposal of 3D printing polymer materials may exacerbate plastic pollution. This article proposes the perspective of a structured risk assessment framework set on particle emissions from industrial 3D printing. It will offer a practical tool to bridge current knowledge gaps and to inform safe practice and policy development, because immediate action is necessary to balance innovation with safety.

Background:Bacterial meningitis (BM) represents the most severe variant of meningitis, with a mortality rate that may reach up to 100% in the absence of appropriate treatment. The success of therapeutic interventions is depends upon prompt and precise diagnostic evaluations. However, there exists a significant deficiency in the literature regarding the diagnostic efficacy within the Ethiopian context. Consequently, this study aims to evaluate and compare the diagnostic precision of microbial culture and polymerase chain reaction (PCR) methodologies in individuals with suspected meningitis in Northwest Ethiopia. Methods:Cerebrospinal fluid (CSF) samples were procured from 400 patients who were clinically suspected of having meningitis and were admitted to the University of Gondar Specialized Hospital (UoGSH), located in Northwest Ethiopia. Real-time PCR, microbial culture, Gram staining, and cell enumeration were conducted at both the UoGSH laboratory and the Armauer Hansen Research Institute in Addis Ababa. Results:Of the total patients enrolled in the study, 58% were male. Clinical manifestations such as fever, headache, and neck stiffness were reported in 94%, 90%, and 81% of the patients, respectively, whereas altered consciousness was recorded in 37% of the cohort. The real-time PCR methodology identified 38 patients (10%) as positive for meningitis, in contrast to microbial culture, which detected only 10 (3%) of these positive cases. The two diagnostic modalities exhibited a correlation coefficient of 0.4 (p < 0.05). Conclusion:The traditional microbiological culture technique, in conjunction with Gram staining, was found to have a limited sensitivity in identifying bacterial meningitis compared with the real-time PCR methodology. Consequently, the integration of molecular approaches with higher sensitivity, such as real-time PCR, facilitates prompt diagnosis and precise treatment, while simultaneously sustaining the overarching meningitis surveillance framework.

The bacterium Neisseria meningitidis causes life-threatening disease worldwide, typically with a clinical presentation of sepsis or meningitis, but can be carried asymptomatically as part of the normal human oropharyngeal microbiota. The aim of this study was to examine N. meningitidis carriage with regard to prevalence, risk factors for carriage, distribution of meningococcal lineages and persistence of meningococcal carriage. Throat samples and data from a self-reported questionnaire were obtained from 2744 university students (median age: 23 years) at a university in Sweden on four occasions during a 12-month period. Meningococcal isolates were characterised using whole-genome sequencing. The carriage rate among the students was 9.1% (319/3488; 95% CI 8.2-10.1). Factors associated with higher carriage rate were age ≤22 years, previous tonsillectomy, cigarette smoking, drinking alcohol and attending parties, pubs and clubs. Female gender and sharing a household with children aged 0-9 years were associated with lower carriage. The most frequent genogroups were capsule null locus (cnl), group B and group Y and the most commonly identified clonal complexes (cc) were cc198 and cc23. Persistent carriage with the same meningococcal strain for 12 months was observed in two students. Follow-up times exceeding 12 months are recommended for future studies investigating long-term carriage of N. meningitidis.

The severity of infectious disease outcomes is dependent on the virulence factors of the pathogen and the host immune response. CARD8 is a major regulator of the innate immune proinflammatory response and has been suggested to modulate the host response to common inflammatory diseases. In the present study, the C10X genetic polymorphism in the CARD8 gene was investigated in relation to bacterial meningitis. A total of 400 clinically suspected meningitis patients hospitalized at the University of Gondar Hospital were enrolled in the study. Cerebrospinal fluid (CSF) and blood samples were collected for laboratory investigations. The collected CSF was cultured, and all the results obtained from the culture were confirmed using direct RT‒PCR. Genotyping of whole-blood samples was performed using a TaqMan assay. The results were compared with apparently healthy controls and with PCR-negative meningitis suspected patients. Of the included patients, 57

The presence of microplastics (MPs) is increasing at a dramatic rate globally, posing risks for exposure and subsequent potential adverse effects on human health. Apart from being physical objects, MP particles contain thousands of plastic-associated chemicals (i.e., monomers, chemical additives, and non-intentionally added substances) captured within the polymer matrix. These chemicals are often migrating from MPs and can be found in various environmental matrices and human food chains; increasing the risks for exposure and health effects. In addition to the physical and chemical attributes of MPs, plastic surfaces effectively bind exogenous chemicals, including environmental pollutants (e.g., heavy metals, persistent organic pollutants). Therefore, MPs can act as vectors of environmental pollution across air, drinking water, and food, further amplifying health risks posed by MP exposure. Critically, fragmentation of plastics in the environment increases the risk for interactions with cells, increases the presence of available surfaces to leach plastic-associated chemicals, and adsorb and transfer environmental pollutants. Hence, this review proposes the so-called triple exposure nexus approach to comprehensively map existing knowledge on interconnected health effects of MP particles, plastic-associated chemicals, and environmental pollutants. Based on the available data, there is a large knowledge gap in regard to the interactions and cumulative health effects of the triple exposure nexus. Each component of the triple nexus is known to induce genotoxicity, inflammation, and endocrine disruption, but knowledge about long-term and inter-individual health effects is lacking. Furthermore, MPs are not readily excreted from organisms after ingestion and they have been found accumulated in human blood, cardiac tissue, placenta, etc. Even though the number of studies on MPs-associated health impacts is increasing rapidly, this review underscores that there is a pressing necessity to achieve an integrated assessment of MPs’ effects on human health in order to address existing and future knowledge gaps.