bims-supasi 2025-11-23 papers

bims-supasi

Biomed News

on Sulfation pathways and signalling

Issue of 2025–11–23
twelve papers selected by
Jonathan Wolf Mueller, University of Birmingham

Chronic Ethanol Drinking Alters Medial Prefrontal Cortex and Nucleus Accumbens Astrocyte Translatome and Extracellular Matrix Glycosaminoglycans.
Modeling highly sulfated infernan.
Heparan sulfate as a central modulator of growth factor dynamics in tissue regeneration.
Cardioprotective potential of sulfated glycosaminoglycan from Amphioctopus marginatus against cardiotoxin-induced toxicity in human iPSC-derived cardiomyocytes.
A high-throughput assay to measure antibodies that block adhesion of Plasmodium falciparum infected erythrocytes to chondroitin sulfate A.
RNA-binding protein PTBP1 mediates HSV-1 attachment and infection through regulation of heparan sulfate 3-O-sulfotransferase gene expression.
Multifunctional Polymeric Nanoparticles Codelivering Paclitaxel and Monoclonal Antibodies for Triple Combination Therapy of Colorectal Cancer.
Evaluation of endogenous steroid sulfates and glucuronides in urine after oral and transdermal administration of testosterone. Part II: Female participants.
Proteoglycofili are glycosaminoglycan-containing fibrillar components released by neutrophils to neutralize bacteria.
Endothelial Glycocalyx Degradation as a Mediator of Neuroinflammation and Cognitive Impairment in Aged Rats: Protective Role of SS-31.
Targeting cancer stem-like cells via cholesterol modulation and ferroptosis induction using a multifunctional nanoplatform to overcome drug resistance.
Prenatal exposure to the pesticides chlorpyrifos and 2,4-Dichlorophenoxyacetic acid is associated with circulating levels of reproductive hormones in healthy infant girls.

bioRxiv. 2025 Oct 02. pii: 2025.09.30.679577. [Epub ahead of print]

Chronic Ethanol Drinking Alters Medial Prefrontal Cortex and Nucleus Accumbens Astrocyte Translatome and Extracellular Matrix Glycosaminoglycans.

Joel G Hashimoto, Angela R Ozburn, Cheryl Reed, Jason Erk, Yuefan Song, Jiyuan Yang, Ke Xia, Fuming Zhang, Yun Yu, Suzanne S Fei, Lina Gao, Robert J Linhardt, Tamara J Phillips, Marina Guizzetti.

Alcohol Use Disorder is a leading preventable cause of morbidity and mortality, yet knowledge of mechanisms driving ethanol-related neuroplasticity remains incomplete. While research has traditionally focused on neuronal signaling, emerging evidence implicates astrocytes in addiction-related adaptations. Here, we investigated the astrocyte-specific molecular consequences of chronic ethanol consumption in the prefrontal cortex and nucleus accumbens, two brain regions critical for executive control and reward processing. Using Translating Ribosome Affinity Purification RNA-seq and bulk RNA-seq in Aldh1l1-EGFP/Rpl10a mice, expressing an EGFP tag on astrocyte ribosomes, we identified hundreds of differentially translated astrocytic genes following chronic continuous two-bottle choice ethanol drinking. Sex-specific analyses revealed greater astrocytic changes in the female PFC and male NAc. Pathway enrichment highlighted extracellular matrix remodeling, synaptic signaling, mitochondrial function, and immune-related pathways. Analyses of individual drinking levels further demonstrated distinct correlations between ethanol intake and astrocytic translation. The major components of the brain extracellular matrix are chondroitin sulfate proteoglycans, produced primarily by astrocytes and covalently bound to chondroitin sulfate glycosaminoglycan chains. Complementary mass spectrometry/liquid chromatography analyses of chondroitin sulfate, heparan sulfate, and hyaluronic acid glycosaminoglycan disaccharides revealed ethanol-induced alterations in chondroitin sulfate glycosaminoglycan sulfation patterns, with additional baseline differences identified between selectively bred high- and low-ethanol preference lines. Together, these findings indicate that astrocytes undergo profound sex- and region-specific adaptations to chronic ethanol, implicating extracellular matrix and glycosaminoglycan remodeling as key risk-factors for and mediators of chronic ethanol-related neuroplasticity.

DOI: https://doi.org/10.1101/2025.09.30.679577
Carbohydr Res. 2025 Nov 17. pii: S0008-6215(25)00385-4. [Epub ahead of print]559 109759

Modeling highly sulfated infernan.

Jacob A Clark, Agata Zykwinska, Sylvia Colliec-Jouault, Sergey A Samsonov.

  Sulfated glycosaminoglycans (GAGs) are a large family of linear and highly negatively charged polysaccharides with many roles in tissue structure and physiology. Interest in glycosaminoglycans and molecules with mimetic properties has led to the discovery of a number of exopolysaccharides (EPS), such as the highly sulfated infernan (InfHS). Understanding of the bioactivity of these molecules depends on their structural features. Here, we constructed and simulated a model of InfHS, aiming to characterize its molecular properties. We observed increased intramolecular interactions as additional repeating units were added to the model, alongside folding of the structure. The branched structure and high sulfation also lead to a more pronounced polarization around the molecule in comparison to linear sulfated glycosaminoglycans. The findings demonstrate the unique properties of InfHS and provide a rationale for understanding its bioactivity.

Keywords:  Glycosaminoglycan mimetics; Marine carbohydrates; Molecular dynamics simulations; Sulfated infernan

DOI:  https://doi.org/10.1016/j.carres.2025.109759
APL Bioeng. 2025 Dec;9(4): 040901

Heparan sulfate as a central modulator of growth factor dynamics in tissue regeneration.

Keyu Tao, Ali Shokoohmand, Simon M Cool, Raymond A A Smith.

Growth factors (GFs) are potent regenerative proteins that modulate biological responses and stimulate tissue repair by activating cell signaling pathways to enhance proliferation, differentiation, and migration. However, the rapid degradation and diffusion characteristics of GFs necessitate the use of supraphysiological doses and/or frequent administrations to maintain therapeutic effects, which can result in reduced efficacy and an increased risk of adverse events. To address these challenges, novel GF delivery systems seek to preserve bioactivity and modulate GF release, ultimately promoting more effective tissue regeneration. Heparan sulfate (HS) is an essential regulator of GF activity, executing molecular recognition and information storage for controlling extracellular matrix remodeling and cellular behavior during tissue development. In this perspective, we summarize the diverse roles of HS in tissue repair, focusing on its interactions with GFs. These include protective functions, the tonic release of GFs, promoting the complexation of GFs with their cognate receptors, and enhancing the activation of intracellular signaling. Finally, we provide perspectives on using HS as a component of novel biomaterials and medical devices for improving tissue regeneration.

DOI: https://doi.org/10.1063/5.0289409
Int J Biol Macromol. 2025 Nov 14. pii: S0141-8130(25)09606-0. [Epub ahead of print] 149049

Cardioprotective potential of sulfated glycosaminoglycan from Amphioctopus marginatus against cardiotoxin-induced toxicity in human iPSC-derived cardiomyocytes.

Silpa Kunnappilly Paulose, Kajal Chakraborty.

  Sulfated glycosaminoglycans, structurally diverse polysaccharides integral to the extracellular matrix of marine animal tissues, are recognized for their pharmacological versatility. This study investigates the cardioprotective potential of AMP-3, a sulfated glycosaminoglycan derived from the marine octopus Amphioctopus marginatus, in mitigating myocardial dysfunction triggered by mitochondrial impairment, calcium homeostasis disruption, and subsequent DNA damage in cardiomyocytes. AMP-3 (100 μg/mL) significantly enhanced myocardial function in human pluripotent stem cell-derived cardiomyocytes (hiPSCs) reducing ATP depletion to 38.7 % and improving mitochondrial membrane potential (Δψm) by fourfold. Additionally, AMP-3 promoted a notable increase in 3D cardiosphere diameter (273 μm), comparable to the standard drug liraglutide (290 μm), and reduced intracellular [Ca2+] levels by 98.6 %, restoring calcium homeostasis. DNA damage was alleviated by 73.7 %, further highlighting its antiapoptotic effects. The structure-activity correlations revealed that higher uronic acid content (⁓23 %), sulfation (⁓10 %) and (1→3) β-glycosidic linkages with C-3 sulfation at β-GlcNAcp in AMP-3 contributed to its prospective cardioprotective effects, as further confirmed by ADME studies. These findings suggest that AMP-3 represents a promising bioactive lead with strong cardioprotective potential and offers a multifaceted therapeutic approach for improving cardiovascular health.

Keywords:  Cardioprotective; Marine octopus Amphioctopus marginatus; Sulfated glycosaminoglycan

DOI:  https://doi.org/10.1016/j.ijbiomac.2025.149049
J Immunol Methods. 2025 Nov 13. pii: S0022-1759(25)00203-0. [Epub ahead of print]545 114003

A high-throughput assay to measure antibodies that block adhesion of Plasmodium falciparum infected erythrocytes to chondroitin sulfate A.

Yvonne Dube, Wina Hasang, Andrew Teo, Mwayiwawo Madanitsa, Morten A Nielsen, Feiko O Ter Kuile, Elizabeth H Aitken, Stephen J Rogerson.

  Placental malaria due to Plasmodium falciparum (Pf) is associated with adverse pregnancy outcomes. Infected erythrocytes (IEs) bind to placental chondroitin sulfate A (CSA) and antibodies that inhibit this adhesion are a potential correlate of protection. We developed a simplified adhesion inhibition assay that uses diaminofluorene to measure haemoglobin release from IEs bound to CSA. Using hyperimmune plasma, the assay demonstrated concentration-dependent inhibition of CSA adhesion. Assay performance was consistent over time with strong reproducibility (r = 0.82), and results correlated with a published assay (r = 0.53). In 466 Malawian pregnant women (321 P. falciparum-infected and 145 uninfected at first antenatal visit), adhesion inhibitory antibodies were significantly higher in mid-pregnancy in infected multigravidae (46.3 % IQR 23.2 %, 74.8 %) compared to infected primigravidae (9.7 % IQR 0 %, 29.3 %, p < 0.001) and in uninfected multigravidae (29.9 % IQR 8.6 %, 54.5 %) than uninfected primigravidae (15.6 % IQR 0 %, 36.4 %, p = 0.04). Similar, significant gravidity-dependent differences were observed at delivery in both infected and uninfected women. Between enrolment and delivery, changes in antibodies were similar in infected and uninfected women. Adhesion inhibitory antibodies protected against placental malaria. Of 162 women with infection at mid-pregnancy, 88 with no placental malaria had more adhesion inhibitory antibody (33.8 %, IQR 1.9 %, 63.2 %) than 74 with past placental malaria (12.5 %, IQR 0 %, 37.8 %; p = 0.002). This low cost, reproducible, and rapid high-throughput adhesion inhibition assay shows promise as a correlate of protection against placental malaria.

Keywords:  Adhesion; Antibodies; Inhibition; Malaria; PfEMP1; Pregnancy; VAR2CSA

DOI:  https://doi.org/10.1016/j.jim.2025.114003
Cell Biosci. 2025 Nov 20. 15(1): 159

RNA-binding protein PTBP1 mediates HSV-1 attachment and infection through regulation of heparan sulfate 3-O-sulfotransferase gene expression.

Weikang Sun, Mengyu Zhang, Ruilin Wang, Jie Yang, Ameena Tur Rasool, Renjie Luo, Xiangdong Liu, Peng Cao, Erguang Li.

   BACKGROUND: The RNA-binding protein polypyrimidine tract-binding protein 1 (PTBP1), also known as heterogeneous nuclear ribonucleoprotein I (hnRNP I), mediates gene expression through splicing regulation. Its role in virus infection is undefined.
RESULTS: We show that genetic ablation of PTBP1 renders cell resistant to herpes simplex virus 1 (HSV-1) infection. HSV-1 utilizes 3-O-sulfated heparan sulfate proteoglycans (HSPGs) for attachment and for infection of epithelial cells. We found that knockout of PTBP1 expression resulted in loss of HS3ST3A1 and HS3ST3B1, heparan sulfate glucosaminyl 3-O-sulfotransferase genes for 3-O-sulfation of the heparan sulfate (HS) chains of HSPGs. Each of the HS3ST3A1/HS3ST3B1 genes is composed of 2 exons separated by an extraordinarily long intron whose removal requires PTBP1-associated looping. We found that PTBP1 interacted with the intronic region of HS3ST3A1/HS3ST3B1 pre-mRNAs and modulated their processing to mRNA. The essential role of PTBP1 in functional HS3ST3A1 expression and in HSV-1 infection was demonstrated by ectopic re-expression in the knockout (ko) cells. In addition, we showed that targeting PTBP1 expression by microRNA mimics reduced disease symptoms in a mouse herpetic stromal keratitis (HSK) model.
CONCLUSIONS: The results demonstrate that PTBP1 mediates HSV-1 infection of epithelial cells through splicing regulation of HS3ST3A1/HS3ST3B1. These studies provide a new area for novel therapeutic strategies through splicing regulation.

Keywords:  Alternative splicing; HS3ST3A1; Herpes simplex virus 1; agomiR-124; hnRNP I

DOI:  https://doi.org/10.1186/s13578-025-01495-7
Biomacromolecules. 2025 Nov 17.

Multifunctional Polymeric Nanoparticles Codelivering Paclitaxel and Monoclonal Antibodies for Triple Combination Therapy of Colorectal Cancer.

Yingchun Zeng, Jingxuan Yang, Ka Wang, Huan Xu, Yue Zheng, Songxin Guo, Jingwen Luo, Sha Liu, Xin Ming, Xue Ying.

Combination therapy is a promising strategy for clinically nonresectable colorectal cancer (CRC) treatment, but its efficacy is compromised by challenges such as cumbersome dosing regimens, poor targeting, high toxicity, and rapid drug clearance. To address these limitations, we designed a functional multitargeted polymeric nanoparticle system (PTX+ATE+BEV-TNPs) modified with octreotide (OCT) and chondroitin sulfate (CS) as targeting ligands to simultaneously encapsulate the chemotherapeutic drug paclitaxel (PTX), the anti-PD-L1 antibody atezolizumab (ATE), and the anti-VEGF antibody bevacizumab (BEV) for triple combination therapy of CRC. The optimized nanoparticles exhibited high endocytosis efficiency and effective lysosome escape, significantly inhibiting the proliferation of tumor cells and promoting their apoptosis. Following systemic administration, PTX+ATE+BEV-TNPs could effectively achieve tumor-targeted codelivery of small-molecule drugs and macromolecular antibodies, promising therapeutic outcomes and displaying good biocompatibility in a CRC mouse model. Overall, PTX+ATE+BEV-TNPs present a viable strategy for CRC and other cancer treatments, demonstrating potential for future translational applications.

DOI: https://doi.org/10.1021/acs.biomac.5c01421
J Steroid Biochem Mol Biol. 2025 Nov 19. pii: S0960-0760(25)00233-X. [Epub ahead of print] 106905

Evaluation of endogenous steroid sulfates and glucuronides in urine after oral and transdermal administration of testosterone. Part II: Female participants.

Sandra Pfeffer, Günter Gmeiner, Nenad Dikic, Marija Andjelkovic, Guro Forsdahl.

  Detecting testosterone (T) doping remains a significant challenge, driving the search for novel biomarkers and advancements in the steroidal Athlete's Biological Passport (ABP). Phase II metabolites of endogenous anabolic androgenic steroids (EAAS) have emerged as promising biomarkers, demonstrating prolonged detection times (DTs) and greater sensitivity compared to conventional biomarkers. Studies involving male participants investigated the effect of intramuscular, oral, and transdermal administration of T on these biomarkers and proposed their integration in future urinary steroid profiling. However, before the inclusion of phase II EAAS metabolites, it is crucial to address a range of possible doping scenarios and the influence of known confounding factors, like ethnicity or sex, on the steroid profile. This study addresses this gap by investigating the impact of oral and transdermal T administration on phase II EAAS metabolites in both men and women. This second part of the study presents the results for female participants, which have not been included in prior research on this topic. Results partially confirm the trends observed in men, with sulfate ratios exhibiting prolonged detection times and higher sensitivity compared to conventional steroid profile biomarkers following multiple oral and transdermal T administration. However, the evaluation in women showed greater variability due to lower steroid concentrations and greater fluctuations influenced by the menstrual cycle. This study provides additional evidence supporting the inclusion of phase II EAAS metabolites for enhanced detection of T doping. Further, it underscores the need for further research to address the unique challenges of female steroid profiling.

Keywords:  Doping; combined ratio; endogenous steroids; phase II metabolites; sex differences; testosterone

DOI:  https://doi.org/10.1016/j.jsbmb.2025.106905
Cell Rep. 2025 Nov 13. pii: S2211-1247(25)01312-9. [Epub ahead of print]44(11): 116541

Proteoglycofili are glycosaminoglycan-containing fibrillar components released by neutrophils to neutralize bacteria.

Antonin C André, Marina V Barroso, Jurate Skerniskyte, Lorine Debande, Vanessa Paul, Nathan Broussaudier, Ahmad Sabbah, Jules Tritschler, Mélina Siegwald, Caroline Ridley, Isabelle Svahn, Amber D Bowler, Valérie Demais, Katia Boniface, Stéphane Rigaud, Jean-Yves Tinevez, Philippe J Sansonetti, Freddy Radtke, Romain R Vivès, David J Thornton, Benoit S Marteyn.

  Here, we uncover that neutrophils release a fibrillar complex, named proteoglycofili (PGF), under hypoxic conditions or upon bacterial infection, using Shigella as a model. PGF are preferentially released by neutrophils over neutrophil extracellular traps upon bacterial infection. PGF are released by living neutrophils, do not contain DNA, are mainly composed of granule proteins, and contain cytokines. We reveal that the fibrillar structure of PGF is sustained by two glycosaminoglycans (GAGs), hyaluronic acid, and chondroitin sulfate, released by neutrophils. We demonstrate that PGF are potent antimicrobials, which degrade virulence factors of Shigella and block its growth, similar to E. coli or Salmonella. GAGs are essential for PGF antimicrobial activity both in vitro and in vivo. Beyond bacterial infections, and reported in a mouse model of colon cancer, our results strongly suggest that GAGs may be released by neutrophils in a broad range of inflammatory diseases.

Keywords:  CP: Immunology; CP: Microbiology; GAG; Shigella; anoxia; antimicrobial; bacteria; chondroitin sulfate; glycosaminoglycan; hyaluronic acid; neutrophils; proteoglycofili

DOI:  https://doi.org/10.1016/j.celrep.2025.116541
Mol Neurobiol. 2025 Nov 17. 63(1): 65

Endothelial Glycocalyx Degradation as a Mediator of Neuroinflammation and Cognitive Impairment in Aged Rats: Protective Role of SS-31.

Min-Hui Kan, Yang Liu, Fan-Qi Meng, Long Fan, Yan Wu, Tian-Long Wang.

  To investigate the role of endothelial glycocalyx (eGC) degradation in mediating the progression from systemic inflammation to neuroinflammation and cognitive impairment in aged rats, and to explore the protective effects of the mitochondrial-targeted peptide SS-31. Aged male Wistar rats (24 months) were assigned to four groups: Vehicle, LPS, LPS + SS-31, and SS-31 alone. Systemic inflammation was induced by intraperitoneal injection of lipopolysaccharide (LPS). SS-31 was administered 30 min prior to LPS injection in the LPS + SS-31 group. Serum levels of eGC degradation products (syndecan-1 [SDC-1], hyaluronic acid [HA], heparan sulfate [HS]) and inflammatory markers (IL-1β, TNF-α) in hippocampus were measured using ELISA. Hippocampal levels of postsynaptic density 95 (PSD-95) were also assessed. LPS-induced systemic inflammation led to significant increases in serum levels of SDC-1, HA, and HS, correlating with elevated hippocampal IL-1β and TNF-α levels and reduced PSD-95 expression. These findings suggest that eGC degradation facilitates the transfer of systemic inflammation to the brain, contributing to neuroinflammation. SS-31 pretreatment attenuated eGC degradation, reduced neuroinflammation, and restored PSD-95 levels, suggesting its potential protective role. eGC degradation is a key intermediary linking systemic inflammation to neuroinflammation and cognitive decline in aged rats. SS-31 may serve as a promising preventive strategy by preserving eGC integrity and mitigating neuroinflammatory processes.

Keywords:  Cognitive dysfunction; Elamipretide; Endothelial glycocalyx; Neuroinflammation; Systemic inflammation

DOI:  https://doi.org/10.1007/s12035-025-05413-3
J Nanobiotechnology. 2025 Nov 19. 23(1): 722

Targeting cancer stem-like cells via cholesterol modulation and ferroptosis induction using a multifunctional nanoplatform to overcome drug resistance.

Leiguang Ye, Jinying Zhu, Xiaoman Wang, Nianhan Chen, Ying Sun, Xuemei Zeng, Shijian Liu, Shuangqian Yan.

  Overcoming therapy resistance in triple-negative breast cancer (TNBC) requires the effective targeting of cancer stem-like cells (CSCs). TNBC is characterized by hyperactivation of the mevalonate pathway, leading to cholesterol accumulation in CSC membranes, which alters membrane properties, enhances stemness, and restricts both drug penetration and lipid peroxidation-a key driver of ferroptosis. Here, we develop Fe/CDP, a nanoparticle with a Fe3O4 core coated with chondroitin sulfate and loaded with pravastatin, a mevalonate pathway inhibitor, and doxorubicin (DOX). In TNBC mouse models, Fe/CDP selectively targets tumors and CSCs via CD44-chondroitin sulfate interactions, enabling localized drug release. Pravastatin suppresses cholesterol biosynthesis, restoring membrane rigidity and fluidity, thereby reducing CSC stemness, disrupting P-glycoprotein function, and downregulating ALDH1, which enhances DOX sensitivity via the EGFR/Src/HMGCR axis. Moreover, cholesterol depletion facilitates lipid peroxidation, synergizing with Fe3O4 to trigger ferroptosis through CoQ10/GPX4/FSP1 downregulation. By eliminating both bulk tumor cells and CSCs, Fe/CDP provides a cholesterol-modulating strategy to overcome TNBC drug resistance.

Keywords:  Cancer stem cell; Cholesterol depletion; Drug resistance; MVA pathway; Membrane modulation

DOI:  https://doi.org/10.1186/s12951-025-03796-y
Reprod Toxicol. 2025 Nov 16. pii: S0890-6238(25)00283-7. [Epub ahead of print]139 109112

Prenatal exposure to the pesticides chlorpyrifos and 2,4-Dichlorophenoxyacetic acid is associated with circulating levels of reproductive hormones in healthy infant girls.

Anna-Patricia Iversen, Jessica Bruun, Lars Christian Lund, Sarah Bakkær Munk Andreasen, Þórhallur Ingi Halldórsson, Anders Juul, Hanne Frederiksen, Anna-Maria Andersson, Casper P Hagen, Flemming Nielsen, Marianne Skovsager Andersen, Tina Kold Jensen, Helle Raun Andersen.

   BACKGROUND: Pesticides are widespread in the environment and suspected endocrine disruptors that may interfere with sex hormones. Following the chlorpyrifos ban in 2020, use of alternative pesticides has increased; 2,4-Dichlorophenoxyacetic acid (2,4-D) remains widely used. This study examined the association between maternal pesticide exposure and pituitary, gonadal, and adrenal hormones in offspring during infancy.
METHODS: We recruited pregnant women from 2010 to 2012 in the Odense Child Cohort, including 489 mother-child pairs. Maternal urinary concentrations of the generic pyrethroid metabolite 3-phenoxybenzoic acid (3-PBA), the chlorpyrifos metabolite 3,5,6-trichloro-2-pyridinol (TCPY), and the herbicide 2,4-D were measured at gestational week 28. Serum concentrations of luteinizing hormone (LH), follicle stimulating hormone (FSH), testosterone (T), estrone (E1), estradiol (E2), 17-hydroxyprogesterone (17-OHP), Androstenedione (Adione), and Dehydroepiandrosterone sulfate (DHEAS) were assessed in infancy. Associations between prenatal pesticide exposure and offspring reproductive hormones (expressed as age- and sex-specific standard deviation (SD) scores) were assessed using multivariate linear regression.
RESULTS: In girls, higher maternal urinary TCPY and 2,4-D concentrations were associated with lower LH (-0.07 SD, 95 % CI: - 0.13; - 0.01 and - 0.06 SD, 95 % CI: - 0.11; - 0.02, per 1 µg/L increase, respectively); there were trends towards associations between 3-PBA, TCPY, 2,4-D and lower LH, FSH, E1 and E2, respectively. No associations were seen in boys.
CONCLUSION: In this low-exposed cohort, prenatal exposure to chlorpyrifos and 2,4-D may affect the reproductive hormones in girls, but not boys, during minipuberty, which may have long-term implications. This is of public health concern given the fact that > 90 % of participants were exposed.

Keywords:  Chlorpyrifos; Cohort study; Herbicide; Minipuberty; Pesticides; Pyrethroids; Reproductive hormones

DOI:  https://doi.org/10.1016/j.reprotox.2025.109112