The utilization of insect protein, specifically black soldier fly larvae meal (BSFLM), offers a viable alternative, sustainable, and nutrient-rich ingredient for pet foods. However, limited information is available on optimal inclusion levels and its functional properties. The objective of the study was to evaluate the effects of incremental levels of BSFLM as a partial or full substitute for chicken by-product meal (CM) in extruded diets. Thirty healthy adult Beagles were assigned to one of three dietary treatments: a control diet with CM, a diet containing 15% BSFLM, and a diet with 30% BSFLM, where CM was partially or fully replaced. Following a 14-day adaptation period on the control diet, the dogs were fed the test diets for 56 days. No significant differences were observed in food intake, fecal output, or apparent total tract digestibility (ATTD) of dry matter, crude protein, and fat across the treatment groups (P > 0.05). However, dogs fed the 30% BSFLM diet showed a reduction in ATTD of organic matter (81.2%) and total dietary fiber (18.9%) compared to the control group (83.2% and 30.3%, respectively; P < 0.05), likely due to the chitin content acting as insoluble fiber. Fecal microbiota analysis showed no changes in alpha diversity; however, unweighted UniFrac analysis revealed significant shifts in beta diversity associated with diet and time. The inclusion of BSFLM-modulated fecal metabolites resulted in lower concentrations of short-chain fatty acids and phenols in the 30% BSFLM group (P < 0.05). Serum biomarkers of oxidative stress and inflammation, including a range of cytokines and antioxidant enzymes, were unaffected by the dietary treatments, indicating no adverse health effects. These findings support the use of BSFLM as a viable and environmentally friendly protein ingredient in canine nutrition, offering a promising alternative to traditional protein sources while contributing to the sustainability of pet food production.

ALI/ARDS are clinical syndromes with diverse etiological origins and are characterized by high mortality rates and a lack of specific therapeutic options. Garlic oil (GO) has been utilized in both culinary and medicinal applications for millennia. However, its complex chemical composition and inherent instability have limited further development and clinical application. We aimed to encapsulate GO within liposomes to increase its solubility and stability. The therapeutic efficacy of GO-loaded liposomes (GO-lips) against LPS-induced ALI was subsequently evaluated in vivo . A novel GO-lip formulation was developed, and its preparation process was optimized to ensure its stability and bioavailability. A murine model of LPS-induced ALI was established. The animals were randomly assigned to the normal control, LPS model, GO treatment, or GO-lip treatment group. Therapeutic outcomes were evaluated by lung tissue histopathology, inflammatory cytokine quantification and oxidative stress biomarker measurement. PCR and molecular dynamics simulations were used to verify the ALI treatment-related pathways influenced by GO-lips. We successfully developed GO-lips using a novel fabrication method. GO-lips demonstrated favorable physicochemical characteristics, with a mean particle diameter of 175 ± 3 nm, a PDI of 0.27 ± 0.02, and an encapsulation efficiency of 70.74 ± 2.11%. Compared with the LPS model group, the GO-lip treatment group exhibited significant protection against LPS-induced ALI. GO-lips demonstrated greater efficacy than free GO, as evidenced by the improved lung histopathology, reduced pulmonary edema, decreased inflammatory responses, and attenuated oxidative stress. PCR analysis demonstrated that GO-lips significantly protect mice primarily via Nrf2 pathway activation. These findings suggest that liposomal encapsulation of GO increases its anti-inflammatory and antioxidant activities, protecting against LPS-induced ALI. This research offers a novel clinical therapeutic approach for ALI and contributes to foundational knowledge supporting the development and utilization of GO-derived formulations.

Academic stress activates physiological responses mediated by the sympathetic-adrenal-medullary axis and the hypothalamic-pituitary-adrenal axis, leading to the release of biomarkers such as cortisol and proinflammatory cytokines. While stress physiology has been extensively studied in clinical populations, few studies have systematically examined the association between academic stress and multiple salivary biomarkers in undergraduates, particularly with attention to sex differences. This cross-sectional study investigated the relationship between self-reported academic stress survey measured via the SISCO inventory and four salivary biomarkers of cellular inflammation: cortisol, interleukin-1β, C-reactive protein, and immunoglobulin A in 81 undergraduates (53 females, 28 males). Biomarker levels were quantified using ELISA, and data were analyzed via multivariate approaches (ANOVA, Pearson correlations, and linear regression modeling). Participants were categorized as low (37%), moderate (35%), and high (28%) stress levels based on SISCO scores. Although no statistically significant associations were found between SISCO scores and individual biomarkers, multivariate analysis revealed a predictive model (R² = 0.14) combining all four biomarkers, with stress level predictions within ±20% of observed values. Males in the high-stress score showed lower cortisol trends but higher proinflammatory markers compared to females, suggesting divergent physiological stress responses by sex. These findings provide preliminary evidence for sex-differential association in self-reported academic stress with biological markers of inflammation, highlighting the potential of biomarker panels rather than single markers to capture the complexity of academic stress. In addition, this study establishes a methodological framework for combining psychometric tools with multi-biomarker analyses in stress research, addressing a critical gap in the literature on academic stress physiology.

Alpha-lipoic acid (ALA) is a naturally occurring organosulfur compound with antioxidant and anti-inflammatory activities. The time-dependent effects of ALA and mechanism of interaction with cystathionine-γ-lyase (CSE—an enzyme responsible for hydrogen sulfide—H2S synthesis) in RAW 264.7 macrophages remain unknown. In this study, we report results supporting the hypothesis that anti-inflammatory effects of ALA are associated with the reduction in CSE expression. To investigate the temporal effect of ALA in lipopolysaccharide (LPS—a potent stimulator of inflammation) treated RAW 264.7 macrophages, ALA was administered 1 h before LPS stimulation and 1, 3, and 6 h post LPS stimulation. Effects of ALA on different inflammatory and oxidative stress biomarkers including tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), monocyte chemoattractant protein-1 (MCP-1), catalase activity (CAT), and malondialdehyde (MDA) levels were investigated. LPS stimulation significantly increased TNF- α, IL-6, MCP-1, MDA levels, and CSE expression and decreased CAT activity compared with the control group (p &lt; 0.05 to 0.0001). ALA treatment at 1000 µM significantly attenuated LPS-stimulated inflammatory response in the macrophages across different time points (p &lt; 0.05 to 0.0001). Furthermore, we found that ALA treatment reduced the expression of CSE in both pre- and post-treated LPS-stimulated macrophages in a time-dependent manner. In conclusion, this study demonstrated for the first time that the protective effects of ALA are dependent on the reduction in CSE expression in LPS-stimulated RAW 264.7 macrophages.

Ecotoxicological effects of cadmium on Echinometra mathaei: Oxidative stress, genotoxicity, and biochemical responses.

Slaughter is a critical phase in aquaculture that can severely compromise both animal welfare and product quality. Stress responses triggered during this stage may accelerate post-mortem biochemical degradation and promote oxidative damage in fish fillets. Essential oils, known for their antioxidant and anti-inflammatory properties, have been proposed as dietary supplements to help mitigate stress and preserve flesh quality. This study investigated the effects of dietary essential oil supplementation and different slaughter methods, air asphyxia and percussion, on stress biomarkers, oxidative processes, and fillet quality in rainbow trout (Oncorhynchus mykiss), both immediately after slaughter and during frozen storage. Air asphyxiation significantly accelerated ATP degradation, increased lipid and protein oxidation products and caused texture loss in fillets assessed immediately post-slaughter. These effects were markedly less pronounced in percussion-slaughtered fish. Lipid mediators such as 12-HpEPE+15-HpEPE and PGD3 + PGE3 were elevated only in asphyxiated fish, providing potential biomarkers for slaughter-induced stress. Proteomic analysis identified several glycolytic enzymes as highly responsive to air asphyxia. Dietary supplementation with essential oils (0.02%) did not mitigate the immediate physiological stress responses to slaughter and was associated with increased muscle protein oxidation at death. However, during 45 days of frozen storage, fillets from fish fed the supplemented diet exhibited delayed progression of oxidative damage, with improved texture and colour retention, especially under high-stress conditions such as air asphyxia. Although dietary essential oils did not reduce slaughter-induced stress, they proved to be an effective strategy for delaying fish fillet degradation and preserving texture and colour during frozen storage. © 2026 The Author(s). Journal of the Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Microplastics (MPs) are prevalent in freshwater systems; consequently, fish ingest them either accidentally or intentionally. Once ingested, MPs can translocate to various organs and cause physiological effects. Most studies have focused on tropical and marine fishes, and many have used mass-based methods that measure exposure only by the total mass of microplastics, ignoring particle number and size. These studies have also rarely examined MP effects or fate after a depuration period, limiting our understanding of MP impacts on temperate fishes, hindering the harmonisation of toxicological studies, and complicating assessments of food safety for cultured and wild fish. This study investigated the physiological impacts of dietary exposure to polystyrene microplastics (PS-MPs; 1–10 µm) in Salmo trutta fed a diet with ~5.4 × 106 PS-MPs g−1 feed for 21 days, followed by a 90-day depuration period. PS-MPs translocation from the intestine to the liver and muscle was investigated. Enzymatic biomarkers of oxidative stress and metabolism were analysed in the liver, digestive enzyme activity was assessed in the intestine, and inflammatory enzyme responses were evaluated in both liver and intestinal tissues. In addition, malondialdehyde (MDA) concentration, an indicator of lipid peroxidation, was quantified in blood, muscle, and liver samples. Results show that 1–5 µm PS-MPs translocated to the liver and muscle, while 10 µm particles largely remained in the intestine, with a small fraction detected in muscle tissue but not in the liver. Most biochemical markers were unaffected; however, both trypsin and peroxidase activities significantly decreased after 21 days, and lipid peroxidation increased in blood following 90 days of depuration. PS-MPs persisted in muscle following 90 days of depuration. These findings demonstrate that dietary exposure to PS-MPs in the size range 1–10 µm leads to selective physiological alterations in S. trutta and results in persistent accumulation of MPs in organs, especially muscle tissue consumed by humans, highlighting a clear concern for food safety.

T50 is the time required for primary calciprotein particles (CPPs) to transform into secondary CPPs in vitro, reflecting serum calcification propensity, and used as a biomarker for calcification stress. Since secondary CPPs induce inflammation and oxidative stress, they may promote atherosclerosis. We investigated whether or not T50 was associated with carotid artery intima-media thickness (IMT). This was a cross-sectional study of 202 health examinees. T50 was measured by the established nephelometric method. Carotid artery IMT was measured by high-resolution ultrasonography. The association between T50 and IMT was evaluated by a multivariable-adjusted linear regression analysis. In a univariate analysis, IMT was not significantly correlated with T50. A multivariable-adjusted linear regression analysis showed that IMT was independently associated with age, sex, diabetes mellitus, dyslipidemia, and fetuin-A but not with T50 in the total subjects. However, when stratified by the estimated glomerular filtration rate (eGFR), T50 was independently and inversely associated with IMT in the subgroup with an eGFR ＜60 mL/min/1.73 m2 (β = -0.418, P = 0.013), whereas it was not in the subgroup with an eGFR ≥ 60 mL/min/1.73 m2. T50 was independently and inversely associated with IMT in health examinees with a reduced kidney function, suggesting a novel link between calcification stress and atherosclerosis, particularly in those with chronic kidney disease.

Introduction Cisplatin (CIS) is a commonly utilized chemotherapeutic agent, but its use is often accompanied by adverse effects such as neurotoxicity and cognitive impairments, collectively referred to as chemobrain. This condition impacts over 70% of cancer survivors, and currently, there are no established therapeutic interventions. This study aimed to evaluate the efficacy of tirzepatide in mitigating the neuropathy effects induced by cisplatin therapy. Methods Forty female Wistar albino rats were divided into four groups of ten: control (untreated), cisplatin (CIS), tirzepatide (TIRZ), and CIS/TIRZ. Treatments were administered intraperitoneally in two injections. The CIS group received cisplatin at a dosage of 5 mg/kg, while tirzepatide was administered at 1.35 mg/kg. In the CIS/TIRZ group, tirzepatide (1.35 mg/kg) was administered prior to cisplatin (5 mg/kg), with a 3-h interval between the two treatments. Post-treatment, behavioral assessments (Y-maze) and oxidative stress biomarkers were evaluated, including enzymatic antioxidants catalase, superoxide dismutase (SOD), and glutathione peroxidase (GPx-1), as well as oxidative damage markers such as reactive oxygen species (ROS) and malondialdehyde (MDA). Results Survival rates were 90% in both the TIRZ and CIS groups, and 70% in the CIS/TIRZ group, whereas all rats in the control group survived. All treatment groups experienced a reduction in body weight compared to the control group. Cisplatin administration resulted in impaired learning and memory in the Y-maze test, which was linked to decreased levels of the antioxidants GPx-1 and catalase, with no alteration in SOD levels. Additionally, ROS and MDA levels were slightly elevated in the CIS and TIRZ groups individually. Although tirzepatide did not ameliorate the memory deficits or antioxidant reductions caused by cisplatin, it did lead to a reduction in ROS and MDA levels. Discussion CIS therapy accelerates memory deficits in female rats by increasing oxidative stress. However, TRIZ did not alleviate the memory deficits or antioxidant reductions, although it did reduce ROS levels.

Several studies have demonstrated that methionine supplementation in fish diets enhances immune status, inflammatory response, and resistance to bacterial infections by modulating for DNA methylation, aminopropylation, and transsulfuration pathways. However, the immunomodulatory effects of methionine in viral infections remain unexplored. This study aimed to evaluate the effect of methionine supplementation on immune modulation and resistance to the viral haemorrhagic septicaemia virus (VHSV) in rainbow trout (Oncorhynchus mykiss). Two diets were formulated and fed to juvenile rainbow trout for four weeks: a control diet (CTRL) with all nutritional requirements, including the amino acid profile required for the species, and a methionine-supplemented diet (MET), containing twice the normal requirement of DL-methionine. After feeding, fish were bath-infected with VHSV, while control fish were exposed to a virus-free bath. Samples were collected at 0 (after feeding trial), 24, 72, and 120 h post-infection for the haematological profile, humoral immune response, oxidative stress, viral load, RNAseq, and gene expression analysis. In both diets, results showed a peak in viral activity at 72 h, followed by a reduction in viral load at 120 h, indicating immune recovery. During the peak of infection, leukocytes, thrombocytes, and monocytes migrated to the infection site, while oxidative stress biomarkers (superoxide dismutase glutathione S-transferase, and glutathione redox ratio) suggested a compromised ability to manage cellular imbalance due to intense viral activity. At 120 h, immune recovery and homeostasis were observed due to an increase in the amount of nitric oxide, GSH/GSSG levels, leukocyte replacement, monocyte influx, and a reduction in the viral load. When focusing on the infection peak, gene ontology (GO) analysis showed several exclusively enriched pathways in the skin and gills of MET-fed fish, driven by the upregulation of several key genes. Genes involved in recognition/signalling, inflammatory response, and other genes with direct antiviral activity, such as TLR3, MYD88, TRAF2, NF-κB, STING, IRF3, -7, VIG1, caspases, cathepsins, and TNF, were observed. Notably, VIG1 (viperin), a key antiviral protein, was significantly upregulated in gills, confirming the modulatory role of methionine in inducing its transcription. Viperin, which harbours an S-adenosyl-L-methionine (SAM) radical domain, is directly related to methionine biosynthesis and plays a critical role in the innate immune response to VHSV infection in rainbow trout. In summary, this study suggests that dietary methionine supplementation can enhance a more robust fish immune response to viral infections, with viperin as a crucial mediator. The improved antiviral readiness observed in MET-fed fish underscores the potential of targeted nutritional adjustments to sustain fish health and welfare in aquaculture.

Efficacy of a short-term physical exercise intervention on stress biomarkers and mental health in adolescents with ADHD: A randomized controlled trial.

A comprehensive screening of different environmental contaminants (total metals, thermostable metal fractions associated with detoxification, and persistent organic pollutants) was conducted in an endangered common thresher shark (Alopias vulpinus ) individual incidentally captured in southeastern Brazil. Stress biomarkers were assessed to evaluate physiological responses, and SDS-PAGE was applied to identify thermostable protein profiles. Most metals were more abundant in total than in thermostable fractions, suggesting partial cellular absorption, likely through compartmentalization in organelles or binding to metallothioneins. Of 25 screened pesticides, only p,p'-DDT and Mirex were detected, with unexpectedly high levels observedin liver. Several low molecular weight PAHs were also found in both tissues, especially muscle, suggesting chronic exposure and bioaccumulation. The SDS-PAGE revealed ~15 kDa bands, consistent with metallothioneins, and higher bands possibly corresponding to matrix metalloproteinases. Poor resolution in the Ampullae of Lorenzini was likely due to high salt content. Antioxidant biomarkers showed tissue-specific patterns, with high H2O2 levels and SOD activity in gills, blood, and brain, suggesting oxidative stress. Further studies are, however, required, as a sample number of one precludes broad conclusions at the species level. Despite this limitation, the study provides valuable preliminary insights and a baseline for Alopias spp. The integrated biochemical and molecular approach applied herein may aid in detecting early physiological stress and sublethal contamination effects and, combined with ecological and life-history data, can inform conservation strategies, such as habitat prioritization, pollutant mitigation, and monitoring programs, to support the long-term survival and population viability of this and other vulnerable shark species.

A Composite Immuno-inflammatory and Oxidative Stress Biomarker Panel for Risk Stratification in Multiple Blood-transfused Patients: A Systematic Review

Background Type 2 diabetes (T2D) is strongly linked to chronic inflammation and oxidative stress, which drive cardiovascular complications. Glucagon-like peptide-1 receptor agonists (GLP-1 RAs) demonstrate cardioprotective benefits that may extend beyond glycemic control, but their effects on key inflammatory and oxidative stress biomarkers compared to other glucose-lowering medications remain inconsistently reported across individual studies. Methods A systematic review and meta-analysis of randomized controlled trials (RCTs) was conducted. Databases were searched for RCTs comparing GLP-1 RAs against other antidiabetic drugs or placebo in adults with T2D, reporting changes in inflammatory biomarkers (C-reactive protein [CRP], interleukin-6 [IL-6], tumor necrosis factor-alpha [TNF-α]) or the oxidative stress marker malondialdehyde (MDA). Data were pooled using a random-effects model, and outcomes were stratified by comparator type (placebo, insulin, other oral antidiabetic drugs [OADs]). Results Forty RCTs (n=6029 participants) were included. GLP-1 RA therapy significantly reduced CRP levels compared to placebo (SMD = -0.59; 95% CI: -0.84 to -0.34) and other OADs (SMD = -1.06; 95% CI: -1.64 to -0.47). A significant reduction in TNF-α was observed versus placebo (SMD = -0.61; 95% CI: -0.89 to -0.32) and oral antidiabetic drugs add on (SMD = -1.62; 95% CI: -2.86 to -0.38). Data for MDA were limited and showed a non-significant trend toward reduction. GLP-1 RAs also significantly reduced IL-6 versus insulin (SMD = -0.24; 95% CI: -0.46 to -0.02). While significant heterogeneity was noted across the analyses, sensitivity analyses confirmed a consistent direction of effect, reinforcing the class-wide anti-inflammatory properties of GLP-1 RAs. Conclusion GLP-1 RAs significantly improve key biomarkers of systemic inflammation (CRP, TNF-α) in patients with T2D compared to various active comparators and placebo. These pleiotropic effects provide a mechanistic rationale for their cardiovascular benefits and support their use as a multifaceted therapeutic strategy in T2D management.

Organic compounds present in environmental pollution are currently regarded as major health threats. Phenanthrene (Phe), a polycyclic aromatic hydrocarbon, impairs testicular function through oxidative stress, leading to the failure of spermatogenesis. This study aimed to explore the potential beneficial effects of photobiomodulation (PBM) on testicular tissue and sperm parameters following Phe exposure in mice. Twenty-four adult male mice, aged 8 weeks, were randomly divided into three groups: control, Phe, and Phe+PBM. In the Phe and Phe+PBM groups, mice received Phe (500 ng/kg) via gavage every 48 hours for 5 weeks. Following Phe exposure, the testes of the Phe+PBM mice were irradiated with laser photons every other day for 35 days. After euthanasia, epididymal tails and testes were collected for molecular and histological analyses. PBM significantly improved sperm count, motility, and viability (p<0.0001). Moreover, reactive oxygen species production, lipid peroxidation, and apoptosis were markedly reduced in the testicular tissue of the laser-treated mice (p<0.0001). Improvements were also observed in seminiferous epithelium thickness and cell distribution following PBM (p<0.0001). Laser therapy significantly mitigates testicular damage from Phe exposure by reducing oxidative stress and apoptosis biomarkers, thereby improving testicular tissue and sperm parameters.

Synergistic effects of metal pollution and habitat degradation from artisanal gold mining drive species-specific oxidative stress and biodiversity loss in a semi-arid river catchment.

Purpose – to evaluate changes in oxidative stress biomarkers in macular tissue and the structural–functional disturbances of the retinal microcirculation system in an experimental model of arterial hypertension. Material and methods The research was conducted on 65 Chinchilla rabbits at the vivarium of Azerbaijan Medical University, where a pharmacological hypertension model was induced using ergometrine maleate for 3, 5, 10, and 15 days. In homogenates prepared from the macular region, major markers of lipid peroxidation (H₂O₂, DC, MDA) and indicators of the antioxidant defense system (protein-SH groups, peroxidase, catalase, TAC) were measured. Results The results showed that oxidative stress progressively intensified in parallel with the increase in arterial pressure. From day 3 onward, the concentrations of H₂O₂ and DC increased noticeably, and this rise became more pronounced on days 5 and 10. By day 15, all lipid peroxidation markers – particularly DC and H₂O₂ – reached their maximum levels compared to the intact state. MDA levels also showed a tendency to rise throughout the model's progression. In contrast, a gradual decline was observed in the markers of the antioxidant defense system. The levels of surface and structural proteinSH groups, as well as peroxidase and catalase, significantly decreased over time. TAC exhibited a comparatively later and milder reduction than the other markers. Overall, a direct relationship was identified between the progression of hypertension, the increase in oxidative stress, and the weakening of the antioxidant defense system. Conclusion The study demonstrates that sustained arterial hypertension enhances oxidative stress in macular tissue, and this process may represent one of the key pathogenic mechanisms underlying retinal microcirculatory dysfunction. Key words: arterial hypertension, oxidative stress, macular tissue, retinal microcirculation

Introduction: While youth with bipolar disorder (BD) spend the majority of the time experiencing symptoms of depression, there are fewer evidence-based options for the pharmacological treatment of bipolar depression versus mania. Given evidence of increased inflammation and oxidative stress in BD, engaging these treatment targets may address underlying pathophysiological mechanisms. We conducted a pilot trial of curcumin, a widely available, safe nutraceutical with anti-inflammatory and antioxidative properties, for the treatment of bipolar depression in youth. Methods: Six participants with bipolar depression were enrolled and received 8 weeks of open-label curcumin. The starting dose was 500 mg daily, then increased to 500 mg twice daily at week 2, then increased to 1000 mg twice daily at weeks 3–8. Symptoms were evaluated with the Children’s Depression Rating Scale–Revised (CDRS-R), Kiddie Schedule for Affective Disorders and Schizophrenia Depression Rating Scale (DRS), and Clinical Global Impression Scale. Treatment response was defined as greater than or equal to 50% reduction in CDRS-R score. Blood biomarkers of inflammation (interferon‐gamma, interleukin-10 [IL-10], IL-8, tumor necrosis factor alpha) and oxidative stress (8-iso-prostaglandin F2alpha [8-ISO], lipid peroxidation [LPO]) were evaluated at baseline, 4 weeks, and 8 weeks. Analyses examined changes in depressive symptoms in relation to changes in biomarkers over time. Results: There were significant reductions in clinical global impression of depression severity [χ 2 (4) = 10.97, p = 0.03, W = 0.46] and overall illness severity [χ 2 (4) = 10.25, p = 0.04, W = 0.43] from baseline to 8 weeks. The most common side effects were related to the central nervous system and gastrointestinal system. From baseline to 4 weeks, greater reduction in CDRS-R scores was associated with greater reduction in 8-ISO ( r = 0.89, p = 0.02), and a greater reduction in DRS scores was associated with a greater reduction in LPO ( r = 0.82, p = 0.05). Conclusions: This trial provides preliminary evidence that the antidepressant effects of curcumin may be associated with its antioxidative properties. However, larger controlled trials are needed to evaluate the efficacy of curcumin for bipolar depression, and to examine oxidative stress markers as predictors and/or mediators of antidepressant effects.

185 Background: Fluoropyrimidines remain the cornerstone of systemic therapy for metastatic colorectal cancer (mCRC), but 5-fluorouracil (5-FU) efficacy is limited by acquired resistance and dose-limiting gastrointestinal toxicity. CF10 is a novel polymeric FdUMP engineered to directly deliver the active thymidylate synthase (TS) and topoisomerase-I (TOP1) inhibitor. Methods: CF10 was studied in clinically relevant preclinical models to determine its antitumor activity, to evaluate its tolerability and examine whether on-target pharmacodynamic (PD) readouts align with efficacy and tolerability signals. A comprehensive set of preclinical systems were used, including patient-derived CRC organoids, isogenic paired parental and 5-FU–resistant cell lines, mouse efficacy/tolerability models and a rat liver-metastasis model. Clinically translatable endpoints- tumor control and survival; body weight; and pharmacodynamic biomarkers of replication stress and DNA damage were measured. Results: In patient derived CRC organoids, CF10 demonstrated superior dose-responsive reductions in size and viability versus control and 5-FU. Consistent with its expected mechanism of action, CF10 activated replication-stress induced DNA damage response pathways, which may provide measurable PD markers for future studies. In three paired parental and 5-FU–resistant cell line models, CF10 effectively killed the 5-FU resistant cell lines, suggesting its potential to overcome drug resistance. In Vivo, CF10 significantly slowed tumor growth and extended survival in mouse models and primary patient derived cell line xenograft (CDX) models. Critically, body weight remained stable and serum chemistries did not worsen relative to 5-FU comparators, providing a strong tolerability signal. Additionally, in a rat liver-metastasis model, a dominant metastatic site, CF10 reduced hepatic tumor burden and improved survival versus control and 5-FU. Conclusions: Overall, CF10 shows preclinical antitumor activity in CRC, including in 5-FU–resistant models and a challenging liver-metastasis setting. These findings are complemented by encouraging tolerability and identification of measurable PD biomarkers. These findings provide promising critical preclinical data to support an early-phase I clinical evaluation of CF10. Median survival (days) in CRC CDX models (TXCC-201, TXCC-199). Median survival (days) CDX model Control 5FU CF10 Log-rank P(CF-10 vs 5-FU) n/arm TXCC201 18 29 43 0.00990 9 TXCC199 24 32 NR 0.0040 8 Medians from Kaplan–Meier; NR = not reached (no death of any mice); P from log-rank (CF-10 vs 5-FU); n/arm shown.

Hair cortisol concentration and adolescent mental health: Insight from the Queensland Twin Adolescent Brain Project.