搜索结果：Current research in toxicology

Oocyte competency is a crucial determinant of fertilisation success and the initial development of embryos in assisted reproductive technologies. The metabolic and biochemical environment of the ovarian follicle is crucial for determining oocyte developmental potential, alongside genetic integrity. The follicular microenvironment includes a complex network of signalling chemicals that regulate mitochondrial activity, steroidogenesis, oxidative balance, and cellular energy metabolism. Recently, metabolic hormones originating from adipose tissue and skeletal muscle, namely, adipokines and myokines, have received considerable focus as crucial regulators of ovarian physiology. Adiponectin, irisin, and the recently identified hormone asprosin have emerged as crucial metabolic regulators influencing granulosa cell activity, mitochondrial bioenergetics, insulin signalling pathways, and redox homeostasis inside the follicular niche. Adiponectin mostly provides metabolic protection by activating AMP-activated protein kinase (AMPK) and improving insulin sensitivity, which in turn enhances mitochondrial efficiency and steroidogenic function in granulosa cells. Irisin, derived from the breakdown of fibronectin type III domain-containing protein 5 (FNDC5), aids the developing oocyte by facilitating mitochondrial biogenesis, augmenting oxidative phosphorylation, and altering cellular defence mechanisms against oxidative stress. Conversely, asprosin has been associated with glucogenic signalling, metabolic stress, and probable mitochondrial malfunction, suggesting a possible relationship between systemic metabolic problems and negative reproductive consequences. Clinical and experimental research indicate that the levels of these metabolic regulators in follicular fluid may correlate with ovarian response, oocyte quality, fertilisation rates, and embryo development during in vitro fertilisation cycles. This review consolidates current molecular, cellular, and clinical information, clarifying the pathways by which adipokines and myokines influence follicular metabolism and impact oocyte competency. Understanding the metabolic connections between systemic endocrine signals and the follicular milieu may provide novel indicators for reproductive prognosis and provide new treatment targets to improve assisted reproduction outcomes.

PM2.5 contributes to lung injury by inducing oxidative stress, inflammatory responses, and apoptosis. Calcitriol exerts a protective role against lung injury by regulating the vitamin D receptor (VDR) and Nrf2 signaling pathways, mitigating PM2.5-induced oxidative stress and inflammation. In this study, we investigated the protective effects of calcitriol against PM2.5-induced apoptosis, oxidative damage, and inflammation in human bronchial epithelial BEAS-2B cells, with a specific focus on the crosstalk between VDR and Nrf2 signaling. BEAS-2B cells were pre-treated with calcitriol (1, 10, 100 nM) for 24 h before PM2.5 exposure (100 µg/mL). Apoptosis, DNA damage, and inflammation were assessed by flow cytometry, ELISA, qRT-PCR, and Western blot analysis. Chromatin immunoprecipitation (ChIP) was performed to evaluate VDR-antioxidant response element (ARE) binding. Calcitriol suppressed apoptotic signaling by reducing p53 phosphorylation and downregulating the mRNA expression of p53 and caspase-3, while also mitigating oxidative DNA damage, as indicated by decreased levels of 8-hydroxy-2'-deoxyguanosine (8-OHdG) in BEAS-2B cells. Additionally, calcitriol suppressed inflammatory responses by downregulating NF-κB activity and the mRNA expression of NF-κB p65 and its downstream pro-inflammatory genes, including IκB-α, TNF-α, and IL-6. Moreover, calcitriol treatment increased VDR protein expression and enhanced Nrf2 activity. ChIP assays demonstrated that calcitriol enhanced VDR binding to AREs, thereby promoting the transcription of key Nrf2-regulated cytoprotective genes, including heme oxygenase-1 (HO-1) and NADPH quinone dehydrogenase 1 (NQO1). These findings provide mechanistic insight into the pharmacological effects of calcitriol, underscoring its potential to alleviate PM2.5-induced cellular injury through VDR-mediated activation of Nrf2 redox signaling.

The increasing use of nanocellulose (NC), including cellulose nanofibrils (CNFs) and cellulose nanocrystals (CNCs), in industrial, biomedical, and consumer products has raised concerns regarding potential inhalation exposure, as these materials contain components within the respirable particle size range (<10 &#xb5;m). Despite expanding applications, data on NC-induced pulmonary and systemic immune effects remain limited. This study investigated the pulmonary and immunotoxic effects of CNF1 (TEMPO-oxidized), CNF2 (mechanically fibrillated), and CNC1 in rats following intratracheal instillation, using multi-walled carbon nanotubes (MWCNTs) as a benchmark. All materials were administered at 2.0&#xa0;mg/kg body weight, with fiber diameters of 14.1-28.2&#xa0;nm and lengths of 0.7-2.2&#xa0;&#xb5;m. At 28 d post-instillation, all NC were phagocytosed by alveolar macrophages. Bronchoalveolar lavage fluid (BALF) and histopathological analyses revealed that CNF2 induced limited inflammation with granuloma formation and minimal BALF changes, whereas CNF1 and CNC1 triggered similar changes in BALF inflammatory markers. Although CNC1 elicited the most notable histopathological changes among NCs, all NC-induced responses were less severe than those caused by MWCNTs. No significant alterations were observed in lymphocyte subsets in the spleen or thymus, indicating minimal systemic immunotoxicity. In vitro assays using NR8383 alveolar macrophages were performed to compare CNC1, sulfuric acid-hydrolyzed CNC2, and desulfurized CNC3. All CNCs were internalized and stimulated pro-inflammatory cytokine production, with responses influenced by surface chemistry despite similar size and morphology. CNC1 and CNC2 exhibited low cytotoxicity following 48h exposure at concentrations up to 100&#xa0;&#x3bc;g/mL. In contrast, CNC3 induced mild to moderate cytotoxicity under the same conditions, upregulated genes linked to inflammatory responses, oxidative stress, apoptosis, and extracellular matrix degradation. These findings reveal that NCs generally exhibit lower pulmonary toxicity than MWCNTs; however, their biological effects are strongly modulated by fiber morphology, surface characteristics, and deposition behavior. To ensure safe use of NCs, comprehensive, material-specific toxicity assessments and standardized evaluation frameworks are essential. In particular, rigorous endotoxin testing and control should be incorporated into future studies to maintain validity and reproducibility of hazard evaluations. Additionally, long-term and repeated exposure models, mechanistic investigations, and case-by-case safety assessments are required, especially for NC variants with limited biodegradability and prolonged pulmonary retention.

The growing prevalence of male infertility has become a significant clinical and public health issue, with environmental exposures increasingly recognized as a major modifiable risk factor. This review synthesizes current evidence within the framework of the "male reproductive exposome", linking lifelong exposure to environmental toxicants-ranging from endocrine-disrupting chemicals to emerging contaminants-to clinically relevant outcomes such as impaired semen parameters, altered reproductive hormone profiles, and an increased risk of testicular dysfunction. We critically evaluate the concept of life-course vulnerability, highlighting how exposures during critical developmental windows-including prenatal, peripubertal, and adult stages-may program distinct pathological trajectories that manifest as reproductive disorders in later life. In addition to classical mechanisms of endocrine disruption, we emphasize oxidative stress and, particularly, epigenetic reprogramming of the germline as key biological pathways contributing to both immediate fertility impairments and potential transgenerational health effects. Furthermore, we discuss the translational importance of these insights, focusing on the development of mechanism-informed biomarker panels for early detection and risk stratification, as well as addressing the persistent challenge of assessing toxicity from complex chemical mixtures. Finally, we underscore the necessity of integrating epidemiological research, mechanistic toxicology, and clinical practice to advance preventive and clinical strategies. This integration requires overcoming methodological challenges in mixture exposure assessment, accelerating biomarker discovery for personalized risk prediction, and formulating evidence-based public health interventions. In a word, this review advocates for a proactive, science-driven approach to mitigate environmental threats to male reproductive health and protect the well-being of future generations.

Due to its ability to inhibit the growth of hepatoma cells, brefeldin A (BFA) has been considered a promising drug candidate for liver cancer. However, there is limited research on its safety profile and potential impacts when administered alone or in combination with other anticancer drugs. To evaluate the safety of BFA in combination with tunicamycin (TM, a candidate anticancer drug) in human normal liver cells (HL-7702) in terms of its ability to induce endoplasmic reticulum (ER) stress and apoptosis. HL-7702 cells were exposed to BFA (0-2.5&#xa0;mg/L) and TM (0-5&#xa0;mg/L), either alone or in combination, for 24&#xa0;h. Cell viability was measured using the CCK-8 assay, and apoptotic rates were determined using flow cytometry. The mRNA and protein levels of key factors related to cell proliferation, ER stress, and apoptosis were determined using quantitative RT-PCR and Western blot, respectively. BFA and TM, either alone or in combination, significantly reduced the viability of HL-7702 cells. BFA alone and BFA&#xa0;+&#xa0;TM combination could weakly induce apoptosis, increase the expression of caspase 12, and reduce the protein level of proliferating cell nuclear antigen (PCNA). BFA alone and BFA&#xa0;+&#xa0;TM combination could significantly increase the mRNA and protein levels of binding immunoglobulin protein (BiP) and activating transcription factor 4 (ATF4), but did not affect the mRNA and protein levels of C/EBP homologous protein (CHOP) and poly (ADP-ribose) polymerase-1 (PARP-1). This study demonstrates that BFA, alone and in combination with TM, exerts mild pro-apoptotic effects on HL-7702 cells, independent of the CHOP and caspase-3 pathways. These findings underscore the necessity of evaluating the potential hepatotoxicity of BFA-based therapies, particularly in combination treatments, to ensure their safe clinical application.

Vinclozolin is a dicarboximide anti-androgenic fungicide that has endocrine-disrupting impact on mammals. Endocrine disruptors are reported to impair mitochondrial function and reduce adenosine triphosphate (ATP) production. Moreover, increasing evidence has linked exposure to endocrine disruptors with metabolic diseases. However, the toxicity of direct exposure to vinclozolin on mitochondrial function remains insufficiently explored. In this study, mitochondria were isolated from mouse liver-one of the primary organs involved in the uptake and processing of toxicants-and directly exposed to vinclozolin. Several markers, including citrate synthase, mitochondrial complex IV (CIV) activity, ATP production, reactive oxygen species/reactive nitrogen species (ROS/RNS) levels, cytochrome c release, glutathione (GSH) levels, and superoxide dismutase (SOD) activity were measured. The results indicated that vinclozolin decreased citrate synthase activity in both sexes and significantly reduced CIV activity only in males. ATP levels showed a decreasing tendency, while ROS/RNS levels showed an increasing tendency particularly in males, without statistical significance. Notably, SOD activity exhibited a sex-dependent increase specifically in females, whereas vinclozolin exposure did not significantly alter GSH levels in either sex. Despite this, basal GSH levels remained significantly higher in females than in males. In both sexes, an increase in cytochrome c release was observed. Collectively, direct mitochondrial exposure to vinclozolin induced dysfunction by impairing energy production, a process mediated by sex-specific antioxidant responses.

Nephrotoxicity is a common side effect of cisplatin (CSP), a widely-used anti-tumor chemotherapy drug. Acute kidney injury (AKI) induced by it is characterized by inflammation, apoptosis, and metabolic reprogramming in proximal tubule (PT) cells. However, the key pathways and upstream transcriptional regulators orchestrating these pathological changes remain poorly understood. This study aimed to identify crucial transcription factors (TFs) involved in CSP-induced AKI and to explore potential therapeutic agents targeting them. An integrative multi-omics analysis of public bulk RNA-seq, proteomic, and single-cell RNA-seq datasets was performed to identify dysregulated pathways and key TFs. The expression of Kr&#xfc;ppel-like factor 15 (KLF15), the lead candidate TF, was validated in a CSP-induced AKI mouse model (20&#xa0;mg/kg intraperitoneal injection for 72&#xa0;h) using RT-qPCR and Western blot. Molecular docking utilizing AutodockVina was employed to virtually screen an FDA-approved drug library for potential compounds targeting KLF15.Transcriptomics and proteomics analyses consistently identified KLF15 as a top-ranked, downregulated TF in CSP induced AKI. Single-cell analysis revealed that PT cells were the most affected, and the loss of Klf15 expression in these cells was strongly associated with a proinflammatory, pro-apoptotic state and suppressed metabolic pathways, including the TCA cycle and fatty acid oxidation. In a murine model of CSP-induced AKI, Klf15 mRNA and KLF15 protein levels were markedly reduced. Furthermore, virtual screening identified 6 drugs, including Simeprevir, Lomitapide, and Avodart, as potential high-affinity compounds targeting human KLF15. In conclusion, our study indicates that the downregulation of KLF15 is a prominent molecular feature in CSP-induced AKI, associated with metabolic failure and injury in proximal tubules.

High pathogenicity avian influenza (HPAI) is a highly infectious and lethal disease of birds that causes systemic symptoms and has been spreading globally, including in Japan. The Okinawa rail (Hypotaenidia okinawae), a flightless bird endemic to Japan, is classified as an endangered species on the Red List. In 2004, the Ministry of the Environment of Japan began implementing a conservation breeding program for Okinawa rails, focusing on maintaining the species' genetic diversity, captive breeding, and reintroduction to the wild. Given the potential for significant losses due to HPAI in Okinawa rails, the establishment of a treatment protocol as a preparedness measure is essential. The aim of this study was to determine an appropriate treatment method for HPAI in Okinawa rails using baloxavir marboxil (BXM), a drug shown to be effective in an avian laboratory model of HPAI virus infection. Single oral administration of BXM at 2.5 or 12.5&#x2009;mg/kg did not produce plasma concentrations sufficient to achieve the expected therapeutic efficacy. Therefore, oral administration was deemed inadequate for generating the desired pharmacological effects. Consequently, subcutaneous administration of BXM to Okinawa rails at a dose of 2.5 or 7.5&#x2009;mg/kg was explored as an alternative protocol, which resulted in higher systemic exposure compared with oral administration. Furthermore, plasma concentrations were maintained at therapeutically relevant levels up to 24 hours after subcutaneous administration at 7.5&#x2009;mg/kg, with mild and reversible injection site irritation the main adverse effect. Based on these results, subcutaneous administration of BXM is proposed as a viable treatment protocol for HPAI in the conservation of endangered Okinawa rails.

Oocyte formation occurs successfully within a meticulously controlled follicular environment characterized by well-documented endocrine, metabolic, and paracrine signals. Yet, the immunological landscape of the follicle and its role in influencing oocyte competency has received less attention in research. Growing research indicates that the ovarian follicle functions as an immunological-active niche necessitating a precise equilibrium between controlled inflammation and targeted immune tolerance. The programmed cell death-1 (PD-1) receptor and its ligand PD-L1 constitute a crucial immune checkpoint pathway, essential for sustaining peripheral immunological tolerance and averting excessive immune activation. Despite their comprehensive research in cancer biology and maternal-fetal interactions, their possible function in the follicular microenvironment remains mostly unexamined. We propose that PD-1/PD-L1 signaling may facilitate the formation of a localized immune-tolerant milieu inside the follicle to safeguard the developing oocyte from inflammatory injury and immune-mediated stress. The disturbance of this suggested equilibrium may lead to a pro-inflammatory follicular environment, compromised granulosa cell function, and modified oocyte maturation, hence affecting fertilization and embryonic developmental potential. In clinical contexts with immunological dysregulation, such as endometriosis, polycystic ovarian syndrome, and unexplained IVF failure, such processes may be especially significant. The purpose of this narrative review is to assimilate the current comprehension of immune regulation in the follicle with the established biology of PD-1/PD-L1 and to investigate a potential correlation between immune checkpoint signaling, oocyte competence, and assisted reproductive outcomes. Considering the follicle as an immune-regulated microenvironment offers a new paradigm for comprehending infertility and identifying novel indicators or therapeutic targets.

As the cosmetic industry replaces traditional animal safety studies with next generation risk assessment approaches, the approach to safety substantiation for peptides used in cosmetic products must also evolve. While the need to provide assurances of safety for local and systemic toxicity endpoints remains the same, adoption of bioinformatic tools developed in the food, agricultural biotechnology, and drug development industries may add to the weight of evidence for the safety substantiation of peptides in cosmetics. Here we review the historical development and safety evaluation of peptides utilized in the cosmetic industry and provide a new safety evaluation framework that incorporates six bioinformatic tools. To test the framework, a variety of peptides (palmitoyl hexapeptide-12, caffeoyl hexapeptide-9, palmitoyl pentapeptide-4, amanitin alpha, conotoxin ArlB, bradykinin, and enkephaline) are evaluated with NCBI BLASTp, ToxinPred3.0, Peptipedia, BIOPEP-UWM, AllerCatPro 2.0, and IEDB bioinformatic tools. The results correctly identified safety concerns (toxins) for amanitin and conotoxin peptides and the biological actions of bradykinin and enkephaline, while palmitoyl hexapeptide-12, caffeoyl hexapeptide-9, and palmitoyl pentapeptide-4 demonstrated sequence homology with extracellular matrix proteins in the skin (collagen, elastin, fibronectin) without the safety concerns of the other peptides. The incorporation of bioinformatic tools into the safety framework provides an additional means to screen for toxins and allergens as well as insights into potential biological activities when sequence homology with existing proteins and peptides occurs. Further testing of the framework by the cosmetic industry is needed to lend support and reveal opportunities for refinements that advance the safety substantiation of peptides.

Microplastics (MPs) are now recognized as pervasive environmental contaminants with emerging evidence of bioaccumulation in the human reproductive system. Recent studies have detected MPs in human follicular fluid, oocytes, placenta, and semen, raising concerns about potential effects on fertility, gamete quality, and embryo development. Experimental data from animal models and in vitro human studies demonstrate that MPs can induce oxidative stress, mitochondrial dysfunction, meiotic spindle disruption, and apoptosis, ultimately impairing fertilization and developmental potential. These findings underscore the urgency of identifying effective strategies to reduce human exposure. Ozone-based water and wastewater treatment technologies are gaining attention as a means to mitigate environmental MPs before human ingestion or contact. Laboratory, pilot-scale, and full-scale plant studies show that ozonation-alone or in combination with granular activated carbon (GAC), coagulation-flocculation, sand filtration, or catalytic oxidation-can remove or degrade a substantial proportion of MPs (up to 96%), with smaller particles often removed more efficiently. Ozone induces surface oxidation and increases hydrophilicity, promoting aggregation and facilitating downstream filtration or sedimentation. While ozonation is less effective than some advanced oxidation processes in complete degradation, it offers the advantage of integration into existing treatment infrastructure. This review synthesizes current knowledge on MPs in ovarian health and the role of ozone-based treatments in limiting exposure. It discusses detection in the ovaries, mechanisms of reproductive toxicity, and the latest engineering strategies for MP removal, with a focus on translational implications for protecting fertility. Despite promising developments, significant gaps remain in linking environmental MP reduction directly to reproductive outcomes. Multidisciplinary collaboration between environmental engineers, reproductive biologists, and clinicians is essential to close these gaps and develop evidence-based public health interventions.

Diisononyl phthalate (DINP) is a high molecular weight phthalate used in commercial products and polyvinyl chloride production. Herein, a systematic evaluation of DINP evidence streams (i.e., human cancer, animal cancer, and mechanistic data) was carried out to inform carcinogenic hazard in humans. Relevant data from peer-reviewed literature and publicly available laboratory reports were extracted and critically appraised. Mechanistic data were organized according to the Key Characteristics of Carcinogens (KCCs) and integrated into key events in rodent cancer modes of action (MoAs). Evidence from epidemiological studies is limited, but does not indicate an association between DINP exposure and cancer, with three studies reporting no association with breast cancer, and one reporting an imprecise increase in prostate cancer risk. Four chronic bioassays demonstrated DINP causes cancer in rodents, with increases in liver tumors in mice and rats, kidney tumors in male F344 rats, and mononuclear cell leukemia (MNCL) in F344 rats. Mechanistic data strongly support that DINP is non-genotoxic (KCC2), and that in rodents DINP induces oxidative stress (KCC5) and alters cell proliferation (KCC10). Multiple evidence stream integration and interpretation support that DINP elicits rodent-specific liver tumors through the peroxisome proliferator-activated receptor alpha, a MoA widely considered to lack human relevance. Likewise, the weak kidney tumor response in male rats was attributed to &#x3b1;2u-globulin nephropathy, a male rat-specific response. MNCL, a common lesion in aging F344 rats, was not considered relevant for predicting human cancer. Together, these data indicate that DINP is unlikely to pose a carcinogenic hazard to humans.

Airway epithelial ferroptosis is critically involved in PM2.5-driven asthmatic airway damage, and our previous studies have demonstrated the protective role of club cell protein 16 (CC16) against fine particulate matter (PM2.5)-induced airway inflammation by inhibiting airway epithelial ferroptosis using C57BL/6&#xa0;J mouse and TC-1 cell model. Nevertheless, it remains unclarified whether CC16 exerts its anti-ferroptotic effects through interacting with E-cadherin, which represents a core research gap absent in our previous studies. Herein, we established PM2.5-intervened asthmatic mice and TC-1 cell models to explore the undiscovered molecular mechanism. Quantitative proteomics combined with bioinformatics screening identified E-cadherin as a pivotal downstream target of CC16. PM2.5 exposure markedly inhibited E-cadherin expression and aggravated epithelial ferroptosis, while CC16 intervention efficiently restored E-cadherin levels, elevated NRF2/GPX4/SLC7A11 expression, suppressed ACSL4, and reduced lipid peroxidation. This study first verified the direct binding interaction between CC16 and E-cadherin via molecular docking, Co-IP and pull-down assays. Furthermore, CDH1 knockdown completely abolished CC16-mediated inhibition of ferroptosis and epithelial injury. Collectively, this work establishes a novel CC16/E-cadherin/ferroptosis signaling axis, demonstrating that E-cadherin is an indispensable mediator for CC16 to relieve PM2.5-triggered airway damage. These findings substantially complement and advance the mechanistic system of CC16-related airway protection, providing novel targets for environmental asthma therapy.

Mutations in RS1 are associated with X-linked retinoschisis (XLRS), a common cause of juvenile macular degeneration in males. Schisis cavities in the central retina of these patients have hampered submacular delivery with conventional adeno-associated viruses (AAVs). Clinical trials employing intravitreally injected AAVs showed a lack of efficacy and inflammation. Here, we demonstrate, in non-human primate (NHP) retina, that AAV.SPR, a laterally spreading capsid, transduced photoreceptors in the macula/fovea without the need for central retinal detachment, enabling transgene expression multiple millimeters beyond the subretinal injection (SRI) bleb margins. Peripheral SRI of AAV.SPR-hGRK1-RS1 resulted in robust and properly localized RS1 expression in NHP fovea. Despite being a secreted protein, biodistribution of RS1 remains confined to the area of AAV-RS1 transduction. Having established feasibility for the approach, we performed preclinical proof-of-concept, safety, and efficacy studies in support of "ATSN-201" (NCT05878860). In RS1KO mice, a "hybrid" efficacy/safety study demonstrated dose-dependent improvements in retinal function and structure and proper localization of RS1 following treatment with ATSN-201. A good laboratory practice (GLP) toxicology study in NHPs established safety at the highest dose evaluated. The enhanced transduction and lateral spreading ability of AAV.SPR make it an attractive option for treating inherited retinal diseases including, but not limited to, XLRS.

Diet has an important impact on intestinal homeostasis, and the establishment of appropriate experimental models to study the effect of food compounds is of interest. The organoid model can be used to check the positive protective role of active food compounds on intestinal tissue. In the current study, mouse intestinal organoids were used to model air-liquid interface (ALI), lipopolysaccharide (LPS)-induced inflammation, and macrophage co-culture-based inflammation modelling. The activity of hesperidin, capsaicin, allicin, and 18&#x3b2;-glycyrrhetinic acid (18&#x3b2;-GA) was determined in organoid culture. Morphology, crypt number, area, and intensity were analyzed. mRNA expression analysis and immunostaining analysis were performed for inflammation and proliferation markers. The ALI model exerted a suitable organoid culture system to mimic intestinal growth based on our results. Hesperidin, capsaicin, and allicin demonstrated positive effects on LPS-induced inflammation. All of the food compounds showed positive effects in macrophage co-culture for organoid structure and growth but not for macrophage proliferation and viability. All compounds reduced the inflammatory gene expression and increased stem cell marker and proliferation-related gene expression in the ALI model. In addition, capsaicin showed positive effects on organoid growth and maturation. This study generated an experimental model system to test food components and might be used in further research.

Chlorine gas (Cl2) is a highly toxic chemical associated with both localized lung injury and systemic health effects. While pulmonary damage has been well characterized, the systemic inflammatory and metabolic responses remain poorly understood. We aimed to define the temporal and multi-organ responses to Cl2 exposure in a murine model, with a focus on identifying spatiotemporal inflammation and its impact on survival and lethality. SKH1 mice were exposed for 10&#xa0;min to varying concentrations of Cl2 (94.4-810&#xa0;ppm, representative of non-lethal, LD10, and LD50 doses) and monitored for respiratory function, perfusion, and acidosis using organ-specific imaging. At multiple time points (40&#xa0;min, 6&#xa0;h, 24&#xa0;h, and 7 d), we measured phosphoproteins, cytokines, chemokines, growth factors, and metabolic hormones in the lungs, heart, cortex, and plasma. Statistical modeling and logistic regression were used to identify biomarkers associated with lethality and survival. We found that lung injury was the primary cause of potential lethality, particularly via early phosphoprotein signaling disruptions. However, survival correlated with early systemic coordination of inflammatory and metabolic signals across organs. Perfusion and acidosis imaging were strongly associated with chemokine and hormone responses. Key survival-associated plasma biomarkers included decreased insulin, increased ghrelin, and decreased eotaxin. While potential lethality from Cl2 exposure is locally driven by pulmonary injury, survival depends on systemic, multi-organ responses that occur rapidly post-exposure. Within this model, our findings identify a potential therapeutic window to enhance survival and suggest candidate biomarkers that may be explored translationally for both triage and treatment of chlorine-related incidents.

Menthol evokes cooling sensations by activating the transient receptor potential melastatin 8 (TRPM8) channel, resulting in relaxing, anti-inflammatory, and analgesic effects when administered through inhalation and topical application. Although the toxicity of menthol is relatively low, the mechanism underlying menthol-induced cytotoxic effects remains unclear. Thus, this study aimed to investigate the cytotoxic effects of menthol in the A549 lung cancer cell line. Menthol induced increases in intracellular Ca2+ concentrations ([Ca2+]i) in distinct modes depending on its concentration. A relatively low concentration (0.3&#xa0;mM) of menthol activated transient receptor potential ankyrin 1 (TRPA1) despite the expression of TRPM8 in A549 cells. A higher concentration (3&#xa0;mM) of menthol nonspecifically induced Ca2+ release from intracellular stores. Menthol inhibited Ca2+-ATPase in the organelle membrane. At 3&#xa0;mM, menthol elicited necrotic cell death accompanied by morphological changes within 60&#xa0;min. This cytotoxicity was not prevented by HC-030031 (a TRPA1 blocker) or BAPTA-AM (an intracellular Ca2+ chelator). Furthermore, the analysis of the cytotoxicity of monoterpene analogs of menthol revealed structure-related activity in menthol-induced cytotoxicity. These findings indicate that menthol-induced cytotoxic effects are concentration-dependent and may provide valuable insights into novel therapeutic strategies for lung cancer.

Fentanyl analogs present a significant global risk to public health and safety due to their high abuse potential and related mortality. However, the absence of a structured research framework focused on their clinical pharmacology and toxicology has led to a scarcity of studies on the mechanisms underlying the toxicity of and addiction to these substances, particularly regarding the rapidly emerging new fentanyl analogs. In this study, we employed a nontargeted metabolomics strategy. By conducting multivariate statistical analysis on ultrahigh performance liquid chromatography-high resolution mass spectrometry (UHPLC-HRMS) data from rat urine and plasma to elucidate the metabolic disruptions induced by fentanyl and carfentanyl. Adult male SD rats were randomly assigned to four groups: low-dose fentanyl group, high-dose fentanyl group, low-dose carfentanyl group, and high-dose carfentanyl group. All groups underwent continuous tail vein injection of toxicants for 5 consecutive days. Plasma and urine samples were collected from the rats before the first administration and at different time points after the last administration, followed by detection and analysis. Specifically, we aimed to elucidate the dose- and time-dependent metabolic toxicity of fentanyl and carfentanyl through a nontargeted metabolomics strategy. Oxidative stress, immunosuppression, and energy dysregulation were identified as core toxic effects of fentanyl and carfentanyl, tightly linked to perturbations in taurine and glutathione pathways, taurine regulating immune function and neuronal homeostasis, glutathione maintaining redox homeostasis. Succinic acid and taurine were confirmed as common biomarkers in urine and plasma. Succinic acid showed decreased plasma levels and increased urine levels, directly indicating fentanyl and carfentanyl-induced energy metabolism impairment. Taurine exhibited similar dysregulation, reflecting drug-induced immunosuppression and neuronal excitability abnormalities. These findings provide critical references and experimental support for studies on fentanyl-related hazards, toxicological mechanisms, and forensic detection.

Areca nut (AN), which is commonly consumed in Southeast Asia, contains bioactive compounds that may influence cellular functions. Accumulating evidence has revealed several health impacts of AN consumption, but the toxicological effects of areca nut extract (ANE) on muscle cells remain largely unexplored. Myogenesis, a critical process for muscle development and regeneration, is closely tied to metabolic activity, which governs the differentiation and function of myocytes. This study aimed to evaluate the effects of ANE on myogenesis in murine C2C12 myoblasts and differentiated myotubes. In ANE-treated C2C12 myoblasts, we observed a significant decrease in the intracellular glutamine level that was accompanied by decreased GSH levels, decreased mTOR signaling, and increased autophagy in myoblasts but not in differentiated myotubes. ANE treatment decreased glutamine and 6-phosphogluconate levels in both myoblasts and myotubes, suggesting the widespread suppression of amino acid and redox-related metabolic pathways. Moreover, ANE significantly altered the metabolomic profile, upregulating the levels of glycolysis and TCA cycle intermediates but reducing ATP levels, indicating impaired energy metabolism in differentiated myotubes. ANE also downregulated the expression of key metabolic genes, including those involved in glycolysis (AKR1B3 and LDHA), glycerol metabolism (GPD1 and GPD2), and nitrogen metabolism (GLUD1, ARG1, GS, and GLS1), indicating that ANE disrupts critical pathways involved in muscle cell metabolism and myogenesis. This study provides new insights into the mechanisms by which AN consumption affects muscle development, emphasizing the need for further research into the dietary and environmental factors influencing myogenesis.