Toxicological responses are inherently spatially heterogeneous, yet conventional analytical strategies generally rely on homogenized samples and therefore provide limited information on where xenobiotics accumulate, which tissue regions or cellular populations are preferentially affected, and how local molecular perturbations relate to pathology and mechanism. Mass spectrometry imaging (MSI) addresses this limitation by enabling label-free, multiplexed, and spatially resolved detection of xenobiotics, metabolites, lipids, peptides, and proteins directly in biological specimens. In this review, we examine the role of MSI in spatial toxicology, with particular emphasis on matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) as a practical core platform for many current tissue-level and emerging cellular-scale toxicology studies. We first discuss why MSI is analytically well suited for spatial toxicology and compare the major MSI platforms in terms of molecular coverage, spatial resolution, and toxicological applicability. We then summarize representative applications at the tissue level, where MSI has already shown clear value in mapping xenobiotic localization, lesion-associated molecular remodeling, and organ-specific toxicity. Next, we discuss cellular-scale MSI and the transition toward true single-cell analysis, emphasizing that current progress is driven more by workflow development, multimodal integration, and computational advances than by routine toxicological deployment. Finally, using per- and polyfluoroalkyl substances as a focused case study, we illustrate how MSI can connect tissue burden, regional bioaccumulation, and localized biochemical response. Overall, MSI is becoming a key analytical framework for mechanistic spatial toxicology, although broader impact will depend on further advances in standardization, annotation confidence, quantification, and multimodal interpretation.
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The accelerating global diversification of food chemicals-encompassing mycotoxins, heavy metals, organophosphate pesticide residues and the increasingly prevalent burden of microplastics and nanoplastics (MNPs)-creates an urgent demand for rapid, human-relevant and ethically acceptable toxicity testing platforms. Conventional animal-based bioassays are insufficient to characterise the thousands of substances awaiting safety evaluation, a deficit particularly acute in tropical food-producing regions including Indonesia where environmental contamination profiles and regulatory infrastructure differ markedly from high-income countries. New approach methodologies (NAMs), comprising validated in vitro cell and organoid systems, high-throughput screening (HTS), multiomics toxicogenomics, in silico modelling and adverse outcome pathway (AOP) frameworks, represent a paradigm shift in food safety toxicology. This review critically appraises current evidence on in vitro NAMs for emerging food contaminants, with emphasis on (i) 2D cell line and 3D organoid/organ-on-chip models for hepatotoxicity, enterotoxicity and nephrotoxicity; (ii) mechanistic pathways-oxidative stress, mitochondrial dysfunction, genotoxicity and intestinal barrier disruption-as interrogated through standardised in vitro endpoints; (iii) AOP frameworks as regulatory bridges between molecular initiating events and adverse outcomes; (iv) high-throughput transcriptomics (HTTr) and Cell Painting integration for mechanism-informed hazard prioritisation; and (v) in vitro-to-in vivo extrapolation (IVIVE) via physiologically based toxicokinetic (PBTK) modelling. Critical gaps in chronic-exposure data, mixture toxicity assessment and BPOM regulatory acceptance are discussed. A novel evidence-based NAM validation roadmap tailored to the Southeast Asian regulatory context is proposed.
Central nervous system (CNS) toxicities remain a major cause of drug attrition and represent a persistent challenge in predicting neurological risk during drug development. Limitations in the predictive resolution and translational relevance of conventional nonclinical paradigms contribute to uncertainty in identifying and interpreting neurotoxicity signals. This manuscript examines key challenges in CNS safety assessment and highlights emerging strategies to improve early detection and prediction of neurological risk. Through a series of case studies, we demonstrate practical approaches for interpreting CNS safety signals and integrating emerging methodologies into nonclinical safety assessment. Examples include sensory and seizure-related endpoints in nonclinical studies and the use of electroencephalography (EEG) to improve detection and characterization of seizure liability. We also highlight the expanding role of advanced sensor technologies and artificial intelligence (AI) in enabling continuous, non-invasive monitoring of animal behavior. In addition, an Integrated Approach to Testing and Assessment (IATA) case study demonstrates how systematic integration of mechanistic data, traditional toxicology findings, and exposure modeling can support regulatory decision-making while aligning with the 3Rs principles (replace, reduce, refine animal testing). Finally, we present regulatory CNS case studies in drug development. Collectively, these approaches enable quantitative assessment of neurological function across circadian cycles, reduce reliance on episodic observer-dependent measurements, and illustrate how integrating refined in vivo methods with New Approach Methodologies (NAMs) and digital technologies can improve prediction of neurological risk and strengthen translation from nonclinical findings to human outcomes in CNS drug development.
To systematically evaluate the dose-dependent ocular safety profile and biological mechanisms of 650 nm repeated red-light (RL) therapy using the chick model. Chicks received monocular 650 nm red-light exposure. Dosimetry was strictly defined as corneal irradiance (power density measured at the corneal surface) under two regimens: a dose-escalation regimen (1, 5, 10, 15 mW/cm2 for 7 days) and a mid-term (5 mW/cm2 for 30 days) regimens (n = 7 per group). Safety assessments included axial length measurements, in vivo macroscopic imaging via swept-source optical coherence tomography and angiography (SS-OCTA), in vivo full-field electroretinography (ffERG), high-resolution ex vivo cellular imaging via full-field OCT (ffOCT), histology/TUNEL assay and untargeted metabolomic profiling coupled with computational toxicology screening. Physiological axial growth was maintained across all irradiances. Retinal architecture, photoreceptor integrity and retinal ganglion cell survival remained intact, with no apoptotic signaling detected. SS-OCTA revealed a significant dose-dependent increase in the choroidal vessel volume per unit area (CVV/a) without accompanying changes in choroidal thickness or vascular index, indicating physiological vascular modulation rather than structural changes. Dual mode of ffOCT demonstrated preserved photoreceptor and retinal ganglion cell layer densities with stable intracellular metabolic motility, ruling out subcellular damage or metabolic stress. ffERG parameters remained stable across all treatment groups. Metabolomic profiling indicated adaptive shifts toward anabolic and bioenergetic pathways, while computational screening detected no enrichment of photochemical or redox-active risk signatures with dose escalation. The chick retina demonstrates a wide biological safety margin for 650 nm repeated red-light exposure, tolerating both supraphysiological irradiance and extended treatment duration without structural, functional or molecular evidence of photochemical injury. These preclinical findings provide evidence evaluating the biological safety margin of repeated 650 nm red light therapy in the retina.
Cadmium (Cd) is classified as a Group 1 carcinogen and constitutes a serious global environmental problem. This study aimed to establish a human colorectal organoid model and to evaluate the feasibility and sensitivity of a luciferase-based ATP bioluminescence assay for detecting toxic responses of colorectal organoids (COs) to Cd exposure. Stem cells were isolated from human colorectal tissue and cultured in a three-dimensional the basement membrane matrix to generate COs. Organoids were treated with a graded series of Cd concentrations for different exposure durations. A luminescent viability reagent was added at an equal volume ratio, gently mixed by rocking at room temperature to promote lysis and reagent penetration, and then incubated to allow the bioluminescent reaction to proceed. Luminescence was recorded in luminescence mode on a multi-mode microplate reader. Dose-response curve fitting was used to calculate the half-maximal inhibitory concentration and to evaluate changes in viability or relative viability. Primary stem cells were successfully isolated from human colorectal tissue and used to generate COs. Cd exposure produced dose-dependent and time-dependent decreases in organoid metabolic activity; dose-response analysis yielded survival curves and IC50 values for the organoids. This study provides methodological guidance and practical insights for utilizing human COs in environmental heavy-metal toxicology studies.
Polybrominated diphenyl ethers (PBDEs) and their metabolite 4-bromodiphenyl ether (BDE3) are pervasive environmental contaminants suspected of endocrine-disrupting effects. This study investigates their inhibitory potential and mechanism of action against human (h3β-HSD1) and rat (r3β-HSD4) placental 3β-hydroxysteroid dehydrogenase, a pivotal enzyme in progesterone biosynthesis. Comprehensive enzymatic analysis revealed that among eight tested PBDE congeners and the structural analog 4-bromobiphenyl (BDP), only BDE3 and BDP exhibited significant inhibition. BDE3 was a potent, mixed/competitive inhibitor with IC50 values of 5.86 μM (h3β-HSD1) and 12.35 μM (r3β-HSD4), significantly more potent than BDP (IC50 ~ 100 μM). Mechanistic studies using pregnenolone as substrate confirmed mixed/competitive inhibition for both compounds, indicating binding at the steroid substrate site, while analysis with NAD⁺ suggested mixed/noncompetitive inhibition. Molecular docking and dynamics simulations corroborated these findings, indicating BDE3 and BDP binding at the steroid/NAD⁺ interface with favorable binding energies, stabilizing the active-site conformation. Network toxicology analysis identified HSD3B1 (encoding h3β-HSD1) as a central hub gene linking PBDE exposure to preeclampsia, enriched in steroidogenesis and estrogen signaling pathways. Single-cell RNA-seq analysis revealed predominant HSD3B1 expression in extravillous and syncytiotrophoblasts, with reduced expression in preeclampsia. In silico virtual knockout of HSD3B1 predicted downstream disruption of cell cycle regulation and steroidogenic pathways. Pharmacokinetic predictions indicated both compounds have high intestinal absorption but potential hepatotoxicity. These results suggest that BDE3 may act as a potent inhibitor of placental 3β-HSD, potentially linking its disruption of progesterone synthesis to adverse pregnancy outcomes.
Eustrongylides spp. are helminths with a multiple-host life cycle that can infect aquatic oligochaetes, various freshwater fish, fish-eating birds, and accidental mammalian hosts, including humans. This work aimed to morphologically and molecularly characterize the larval stages of Eustrongylides spp. isolated from European perch, and to outline the oxidative/nitrosative stress and histoarchitecture changes induced by active larval migration or tissue-encapsulation. Eighty Perca fluviatilis were collected from the Volga River (Astrakhan, Russia). Thirty-seven were tentatively grouped as infected based on the grossly visible Eustrongylides larvae in the oral cavity, gills, axial muscles, and internal viscera. The nematodes were examined microscopically and characterized at the molecular level by PCR amplification of nuclear small-subunit ribosomal RNA (nSSU-rRNA) gene fragments, followed by sequencing and phylogenetic analysis. Affected organs were evaluated for redox/ nitrosative status and histopathological tissue injury. Fifteen healthy fish were selected as controls. Morphological examination identified the isolated nematodes as Eustrongylides sp., which were subsequently identified as Eustrongylides excisus (E. excisus) by molecular analysis, contributing to an infection rate of 46.25% (37/80). E. excisus was identified based on 100% sequence identity with available GenBank references, although the 18S rRNA marker provides limited species-level identification. Our findings demonstrate that E. excisus infection triggers a significant shift in the host's oxidative and nitrosative burden. This oxidative-inflammatory response leads to marked pathological changes in affected organs, including blood vessel dilation, mononuclear cell infiltration, vacuolar degeneration, and necrosis. Ultimately, these larvae induce severe cellular damage in fish muscle and visceral organs, disrupting cellular integrity essential to normal physiological function.
Herein, we report the synthesis of new 1,4-benzothiazine-1,1-dioxide derivatives bearing bis(1,2,3-triazole) moieties via click chemistry in good to high yields and fully characterized by 1H NMR, 13C NMR and HRMS. The compounds were evaluated using a combined in silico approach, including physicochemical, pharmacokinetic and toxicity analysis, molecular docking and molecular dynamics studies. Drug-like properties of all compounds were favourable. Molecular docking against SRC kinase suggested binding affinities ranging from -7.14 to -8.13 kcal/mol. Compound 11 displayed the most favorable docking score (-8.13 kcal/mol), comparable to that of bosutinib (-8.54 kcal/mol), and formed key interactions within the active site. This finding was consistent with the 100 ns molecular dynamics simulations, which indicated that the SRC-compound 11 complex remained structurally stable and maintained key interactions with residues such as Met341, Tyr340 and Lys295. Despite showing conformational flexibility than bosutinib, compound 11 maintained stable binding throughout the simulation. Overall, compound 11 showed favorable predicted binding toward SRC kinase, suggesting that these bis-triazole derivatives may represent potential leads for future anticancer investigation.
Microplastics (MPs) are emerging environmental pollutants that have received increasing attention in recent years. However, data on their potential risks to mammalian species remain limited. In this study, female rats were exposed via oral gavage to pristine (PPS) and fluorescent polystyrene (FPS) particles (5 μm diameter) over four estrous cycles. FPS-MPs were detected in hepatic tissue (133 ± 35 particles), confirming their translocation. Exposure to PPS-MPs increased superoxide dismutase (SOD) (2.58-fold) and catalase (CAT) activity (1.36-fold), while reducing protein sulfhydryl levels (PSH) (0.35-fold), indicating oxidative stress. Histological analysis and quantitative assessment revealed significant alterations, including significant decrease in glycogen content (2.4- fold). Immunofluorescence analysis showed a pronounced increase in α-tubulin and a decrease in DAAM-1 signals, a formin protein essential for actin filament assembly and cytoskeletal organization, suggesting impaired cytoskeletal dynamics. These observations were further supported by molecular analyses, which showed α-tubulin overexpression (3.35-fold) and reduced DAAM-1 expression (0.5-fold). Our findings indicate that exposure to environmentally relevant concentrations of MPs induces early hepatocellular alterations, suggesting that oxidative stress and cytoskeletal remodeling may be central mechanisms underlying MP-induced hepatotoxicity. These findings should be interpreted with caution, as the study used only female rats, a single dose, particle size, and polymer type, with a short exposure duration, and it should be noted that FPS-MPs were used only to track distribution, whereas toxicity assessments were performed with non-fluorescent particles.
This systematic review evaluated the effects of fructose intake on neuroinflammatory markers in rodent models. The search terms Fructose AND neuroinflammation OR Neurodegeneration OR chemokines OR interleukins OR microglia OR behaviour OR memory OR cognition were used in Google Scholar, Scopus and Web of Science. Thirteen animal studies investigating fructose-induced neuroinflammation that matched the eligibility criteria were included in the study. Across the studies, 16 inflammatory markers were identified as significantly altered following exposure to fructose. The findings consistently demonstrated elevated expression of pro-inflammatory cytokines, TNF-α, IL-6, and IL-1β, following fructose administration. Fructose consumption also dysregulated MCP-1, fractalkine, and CX3CR1 levels, thereby promoting inflammatory signalling and microglial activation. Furthermore, fructose exposure significantly increased IBA-1 and CD11b, indicating sustained neuroimmune activation. Alterations in important inflammatory pathways involving TLR4, NLRP3, NF-κB, MyD88, iNOS, and cyclooxygenases (COX-1 and COX-2) were also observed. In contrast, expression of the anti-inflammatory regulator peroxisome proliferator-activated receptor gamma (PPARγ) was reduced after fructose treatment. Overall, the findings suggest that chronic fructose consumption induces neuroinflammation through multiple inflammatory and immune-related mechanisms in the brain. These effects appear to be dose- and duration-dependent and may contribute significantly to neurodegeneration and cognitive impairment.
The current study devised and synthesized a novel class of pyrazole derivatives based on indole as possible inhibitors of cyclin-dependent kinase-2 (CDK-2). 1H NMR, 13C NMR, NOESY, HMQC, and elemental analysis were used to confirm the structural integrity of the synthesized compounds. Promising CDK-2 inhibitory activity was observed in biological assays, and numerous compounds exhibited sub-micromolar IC50 values. Compound 5 d outperformed the reference inhibitor Roscovitine (IC50 = 0.716 µM) as the most potent inhibitor (IC50 = 0.536 µM), followed by compound 9 g (IC50 = 0.675 µM). SAR analysis showed that the observed activity was significantly influenced by the electronic nature of the added substituents as well as the orientation of the indole bond, with brominated derivatives exhibiting greater potency. The antiproliferative activity of the most potent compounds against the cancer cell lines HepG2, HCT-116, and MCF-7 was further assessed. In addition to having an enhanced selectivity index for normal MCF-10A cells (SI = 8.00 vs. 4.96 for Roscovitine), compound 5 d had the greatest activity against MCF-7 cells (IC50 = 6.78 µM), surpassing Roscovitine (IC50 = 8.11 µM). According to mechanistic investigations, compound 5 d significantly reduced the S-phase population, markedly promoted apoptosis, and caused G1 and G2/M cell-cycle arrest. Additionally, the consistent binding of compound 5 d within the ATP-binding pocket of CDK-2 was confirmed by molecular docking and molecular dynamics simulations, and attractive drug-like and pharmacokinetic features, similar to those of Roscovitine, were demonstrated by in silico ADMET predictions. All of these results point to compound 5 d as a promising lead scaffold for developing potent CDK-2-targeted anticancer agents.
Liver regeneration after hepatectomy is a challenge in patients with metabolic dysfunction-associated steatohepatitis (MASH), as well as small-for-size syndrome and ex vivo organogenesis. Endogenous lipid metabolites promoting liver regeneration are poorly understood. A 70% partial hepatectomy (PHx) was performed in mice with healthy and MASH liver. Biomarkers promoting liver regeneration were explored using hepatic lipidomics followed by structural identification. The functional validation was performed in in vivo and in vitro models. Liver regeneration post-PHx peaked earlier in the MASH liver than that in the healthy liver. Phosphatidylcholine (PC) (16:0/20:4) increased in both MASH liver and non-MASH liver, and it promoted the proliferation of Hepa 1-6, Huh-7, human liver organoids, and mouse PHx models. In patients with metabolic dysfunction-associated steatotic liver disease (MASLD) and those undergoing hepatectomy, the serum PC (16:0/20:4) also increased, supporting the findings in this work. PC (16:0/20:4) is an endogenous lipid biomarker promoting liver regeneration which increased during hepatectomy in healthy and MASH liver.
Distinguishing peripheral blood (PB) from menstrual blood (MB) is critical in forensic body fluid identification, particularly in sexual assault and violent crime investigations. The miR-451a/miR-21-5p ratio has been proposed as a discriminatory marker, but its stability under environmentally relevant temperatures has not been systematically evaluated. To assess the effect of three temperatures (4 °C, 24 °C, and 40 °C) on the discriminatory performance of the miR-451a/miR-21-5p ratio for PB-MB differentiation, and to compare ratio-based performance against individual miRNA analysis. Forty blood samples (20 PB, 20 MB) were collected from healthy female volunteers. Total RNA was extracted, and miR-451a and miR-21-5p levels were quantified by RT-qPCR after 24 h of exposure to each temperature. The ratio was significantly higher in PB than in MB at all temperatures (P < 0.001). Individual miRNA analysis was less reliable: miR-451a significantly differentiated PB from MB only at 4 °C and 40 °C, while miR-21-5p did not differentiate between blood types at any temperature. Receiver operating characteristic (ROC) analysis identified cut-offs of 1.06, 1.02, and 0.765 at 4 °C, 24 °C, and 40 °C (all P < 0.001), with sensitivity/specificity of 70/90%, 100/100%, and 100/100%, respectively. A universal, temperature-independent cut-off (<0.900) also achieved acceptable discrimination (AUC 0.927; sensitivity 96.7%, specificity 86.7%). The miR-451a/miR-21-5p ratio maintained discriminatory performance across all temperatures and outperformed individual miRNA markers. However, the observed differences reflect the combined effect of temperature and storage format rather than temperature alone; therefore, future studies under identical dried conditions are needed to isolate the temperature effect.
Leishmaniasis is a vector-borne parasitic disease transmitted by female sandflies of the genus Phlebotomus and affects approximately 1.2 million people annually in more than 90 countries worldwide. Morocco remains one of the most affected countries in North Africa, where the disease continues to represent a major public health concern. This study aimed to analyze the spatial and temporal trends of leishmaniasis incidence across all municipalities of Errachidia Province between 2010 and 2025 and to assess the influence of ecological, demographic, and socio-economic factors on its distribution. Also, the study aimed to predict the monthly CL cases using the Seasonal Autoregressive Integrated Moving Average (SARIMA) model. Epidemiological data on parasitologically confirmed cases obtained from the Errachidia Provincial Health Delegation were processed using geographic information system (GIS) tools and statistical methods to explore spatial patterns and correlations with environmental and socio-demographic variables. The models are trained and evaluated using monthly CL cases collected from 2010 to 2025, with the optimal model selected based on Akaike Information Criterion (AIC). During 16-years study period, 7034 cases were recorded in Errachidia Province, with the highest incidence reported in 2010 (860 cases per 100,000 inhabitants). Overall, the incidence showed marked temporal fluctuations but demonstrated a general decreasing trend over the study period. A pronounced spatial heterogeneity was observed between rural municipalities (Sid Ali, Melaab, and Ferkla) and urban municipalities (Errachidia, Arfoud, and Goulmima) (p < 0.01). The disease was slightly more frequent in females (54.41%) than in males (45.59%), and a significant difference was observed among age groups (p = 0.019), with the 0-9 and 10-19-year groups being the most affected. Seasonal analysis revealed a peak incidence during winter. In addition, higher incidence was associated with low- to medium-altitude municipalities, while no significant association was observed with poverty or vulnerability indices. The SARIMA (0,0,1)12 model demonstrated the best predictive performance. These findings highlight the heterogeneity and ecological determinants of leishmaniasis in southeastern Morocco and may support targeted surveillance and control strategies in high-risk areas.
Neutrophils are the most abundant circulating leukocytes and play a central role in innate immunity. As in vitro functional assays (NETosis, degranulation, phagocytosis, etc.) depend on cell viability, purity and basal activation, it is essential to validate neutrophil suspensions before experimental use. To characterize isolated neutrophils using an in-house flow cytometry workflow for assessing viability, purity, and activation, and to establish a reference interval (RI) for neutrophil basal activation. Twenty healthy volunteers were recruited, and neutrophils were isolated from whole blood by negative selection. Cell viability was determined by propidium iodide staining. Leukocyte and neutrophil percentage were evaluated using conjugated antibodies against CD45 and CD15 respectively. Basal activation was assessed by measuring mean fluorescence intensity (MFI) using a conjugated antibody against CD66b. To validate the RI of basal activation, neutrophils were also stimulated with TNF. RI were calculated as mean ± 2 standard deviations (SD). Neutrophil suspensions displayed high quality across all criteria. Viability averaged 99.4% (SD =0.4). Leukocyte purity reached 99.3% (SD =0.4), and neutrophil purity was elevated, with a mean of 98.6% (SD = 0.5). Isolated neutrophils presented a mean CD66b MFI of 37.4 (SD =10.8), and CD66b expression increased significantly following TNF stimulation. The calculated RI was 37.4 ± 21.6. This study demonstrated that neutrophils isolated by negative selection displayed high viability, purity, and minimal activation. It also allowed the definition of a RI for basal activation which could serve as objective criterion to validate neutrophil suspensions for functional assays.
The NAT2 pharmacogene is essential in drug metabolism, particularly for aromatic amines and hydrazines. Genetic variations in NAT2 categorize individuals as rapid, intermediate, or poor metabolizers based on their acetylation capacity to inform dosing guidelines. Genotyping and short-read sequencing allele calling methodologies are unable to phase variants, leading to ambiguous diplotype and phenotype calls while long-read sequencing technologies not only provide variant detection but also variant phasing to unequivocally establish diplotype. This study utilized long-read sequencing to analyze NAT2 genetic variation in a large cohort of 1828 long-read and 662 paired short-read patient samples from the Genomic Answers for Kids program, comparing call accuracy with short-reads and simulated SNP panel tests. A custom tool, staR-NAT2, identified 11 new star alleles and four new suballeles. Long-reads demonstrated high diplotype accuracy (> 99%) with pb-StarPhase and Aldy, excluding novel haplotypes, while short-reads showed a significant reduction to 64% in diplotype concordance, but only affecting 4% of phenotype calls. Despite poor diplotype resolution and accuracy compared with long-read data, short-read sequencing and a 4-SNP panel accurately predicted phenotype for over 95% of subjects. However, the 4-SNP and 5-SNP panels both resulted in discordant or ambiguous phenotypes for non-white individuals at 4.5% and 25%, respectively. Additionally, population-specific alleles were identified among 1144 unrelated individuals, including NAT2*14 (8.9%) and *43 (3.57%) in Blacks and *7 (7.35%) in Hispanics. These findings underscore the limitations of short-read sequencing and support long-read sequencing as a robust approach for accurate and equitable NAT2 pharmacogenetic testing to guide individualized drug therapy.
Knowledge of Haemaphysalis ticks infesting domestic and wild animals remains scarce in northern Kerala, India. A systematic study was undertaken to identify and characterize Haemaphysalis tick species infesting domestic and wild animals in northern Kerala, India. They were further characterized using mitochondrial 16S rRNA, nuclear 18S rRNA and mitochondrial cytochrome c oxidase subunit 1 (MT-COI) genes and the phylogenetic analysis was performed using the generated sequences. In this study, H. bispinosa, H. spinigera, H. turturis, H. minuta, and H. sambar were recovered from wild animals, and H. bispinosa, H. spinigera and H. turturis were identified in domestic animals. The most predominant haemaphysalid ticks infesting goats was H. bispinosa (70.44%), followed by H. turturis (1.25%); while in cattle, it was H. bispinosa (78.88%), followed by H. spinigera (9.31%). The interspecific genetic distance of all the Haemaphysalis sp. studied based on MT-COI ranged from 9.52 to 26.73%, exceeding the delimiting genetic distance. In addition, the rediscovery and redescription of H. sambarHoogstraal, 1971, from a new host, the Bengal tiger in Kerala, India, previously known only from a single male specimen, is also reported.
The heterogeneity in associations between circulating fatty acids (FA) and mortality remained underexplored. Proteomics can profile the human physiological status. This study aimed to estimate interactions between FA and proteins in relation to mortality. We randomly divided 30,190 UK Biobank participants into train and test datasets. Multivariable Cox regression was utilized to assess the associations between FA and all-cause mortality and to explore proteome-wide interactions of FA in relation to mortality. Subgroup analyses were conducted to examine heterogeneity across varied protein levels. We also explored interactions between proteins and FA in relation to cause-specific mortality. We documented 3,345 deaths during 13.9 years of follow-up. MUFA-pct, Non-LA Omega-6 pct, Omega-6/Omega-3 ratio and SFA-pct were positively associated with all-cause mortality, while PUFA-pct, DHA-pct, LA-pct, and Omega-3-pct were negatively associated. We identified several robust interactions of proteins with MUFA-pct (n = 3), Omega-3-pct (n = 4), and Omega-6/Omega-3 ratio (n = 2). In subgroup analyses, individuals with high-level TNFRSF1B, MMP10, and CRHBP had higher all-cause mortality risks associated with MUFA-pct, while protective associations between Omega-3-pct and all-cause mortality were stronger among individuals with high-level TSPAN8, PLAU, ITGA5, and CEACAM1. Moreover, participants with high-level TSPAN8 and PLAU had higher risks of all-cause mortality with Omega-6/Omega-3 ratio. For cause-specific mortality, interaction and subgroup results were largely consistent with those of all-cause mortality. Our findings can provide new insights into heterogeneity in FA-mortality associations and highlight potential protein targets for personalized interventions across individuals with different physiological status.