Dibutyl phthalate disrupts [Ca2+]i, reactive oxygen species, [pH]i, protein kinases and mitochondrial activity, impairing sperm function.

To explore the mechanism of sperm dysfunction caused by dibutyl phthalate (DBP), the effects of DBP on intracellular [Ca2+] and [pH], reactive oxygen species (ROS), lipid peroxidation (LPO), mitochondrial permeability transition pore (mPTP) opening, mitochondrial membrane potential (MMP), adenosine triphosphate (ATP) levels, phosphorylation of protein kinase A (PKA) substrate proteins and phosphotyrosine (p-Tyr) proteins, sperm motility, spontaneous acrosome reaction, and tail bending were examined in mouse spermatozoa. At 100 µg/mL, DBP significantly increased tail bending and [Ca2+]i. Interestingly, DBP showed biphasic effects on [pH]i. DBP at 10-100 µg/mL significantly decreased sperm motility. Similarly, Ca2+ ionophore A23187 decreased [pH]i sperm motility, suggesting that DBP-induced excessive [Ca2+]i decreased sperm motility. DBP significantly increased ROS and LPO. DBP at 100 µg/mL significantly decreased mPTP closing, MMP, and ATP levels in spermatozoa, as did H2O2, indicative of ROS-mediated mitochondrial dysfunction caused by DBP. DBP as well as H2O2 increased p-Tyr sperm proteins and phosphorylated PKA substrate sperm proteins. DBP at 1-10 µg/mL significantly increased the spontaneous acrosome reaction, suggesting that DBP can activate sperm capacitation. Altogether, DBP showed a biphasic effect on intracellular signaling in spermatozoa. At concentrations relevant to seminal ortho-phthalate levels, DBP activates [pH]i, protein tyrosine kinases and PKA via physiological levels of ROS generation, potentiating sperm capacitation. DBP at high doses excessively raises [Ca2+]i and ROS and disrupts [pH]i, impairing the mitochondrial function, tail structural integrity, and sperm motility.

Colonization of phthalate-degrading endophytic bacterial consortium altered bacterial community and enzyme activity in plants.

Phthalates (PAEs) are widely distributed hazardous organic compounds that pose threats to ecosystems and human health. Endophytic bacteria can effectively eliminate PAEs contamination risk. However, limited information is available regarding the impact of endophytic bacterial colonization on bacterial communities within plants. In this study, the endophytic bacterial consortium EN was colonized in lettuce by seed soaking, root irrigation, leaf spraying, and combined spraying-irrigation, resulting in a marked improvement in plant growth. The findings revealed that consortium EN colonization through combined spraying-irrigation exhibited superior degradation capability with 40.54% PAEs removal from soil. Meanwhile, the residual PAEs in lettuce decreased by 94.05% compared with the uninoculated treatment. High-throughput sequencing analysis indicated that colonization of consortium EN altered the bacterial community in lettuce. Specifically, the relative abundance of the dominant genus Pseudomonas was significantly higher than that in the uninoculated control (P < 0.01). Additionally, colonization enhanced the activities of peroxidase and catalase in lettuce, thereby improving plant resistance. This work offers a theoretical foundation for comprehending the mechanism underlying the bioremediation of PAEs contamination by endophytic bacteria in soil-plant system.

Impact of diethyl phthalate on freshwater planarian behaviour, regeneration, and antioxidant defence.

Diethyl phthalate (DEP) has been widely used as a plasticiser in various consumer products, including cosmetics, personal care items, and pharmaceuticals, and recent studies reported a higher abundance of this priority phthalate in the aquatic environment. DEP is a potential endocrine disruptor, affecting immune systems in humans and wildlife even at low-level chronic exposure. As concern over phthalates increases globally, regulatory bodies focus more on their environmental impact. However, limited research is available, particularly using model organisms like planarians. Planarians are ideal for toxicological studies and may provide insightful information on pollutants' neurotoxic, developmental, and ecological effects, especially in freshwater environments where planarians play a vital role in ecosystem balance. Therefore, the objective of the current study was to examine the toxicity of DEP using the freshwater Dugesia sp., as an experimental animal. The LC50 for the test organism was calculated using DEP concentrations of 800, 400, 200, 100, and 50 µM, with an estimated LC50 of 357.24 µM. Furthermore, planarians were exposed to sub-lethal DEP concentration (178.62 µM) for one day as well as eight days to evaluate the impact of DEP on planarian locomotion, feeding behaviour, and regeneration ability. At sub-lethal concentration, locomotion and feeding ability were decreased, and regeneration was delayed. Furthermore, neuro-transmittance in planaria was altered by sub-lethal DEP concentration, as indicated by a reduced acetylcholinesterase (AChE) activity. DEP exposure induced oxidative damage in the tested planarians as shown by a marked increase in stress biomarkers, including lipid peroxidation levels and antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT), peroxidase (POX), and glutathione S-transferase (GST). Our study revealed that DEP exposure may prove fatal to freshwater organisms, such as planarians. The observed alterations in behaviour and regeneration ability demonstrate the severity of the effects exerted by DEP as a toxicant in aquatic ecosystems, thereby indicating the need to restrict its usage to protect aquatic environments.

An ultrasensitive free-of-electronic sacrificial agent photoelectrochemical aptasensor for the detection of dibutyl phthalate based on Z-scheme p-n Bi-doped BiOI/Bi2S3 heterojunction.

Dibutyl phthalate (DBP), a common and outstanding plasticizer, exhibits estrogenic, mutagenic, carcinogenic, and teratogenic properties. It is easily liberated from plastic materials and pollutes aquatic ecosystems, endangering human health. Therefore, highly sensitive and selective DBP detection methods are necessary. In this work, a free-of-electronic sacrificial agent photoelectrochemical (PEC) aptasensor for DBP detection was constructed using a novel Z-scheme Bi-doped BiOI/Bi2S3 (Bi-BIS) p-n heterojunction. The Bi-BIS composites had higher visible-light absorption, charge transfer, and separation efficiency. This is attributed to the synergistic effect of the formation of Z-scheme p-n heterojunction between BiOI and Bi2S3, the plasma resonance effect of metallic Bi and photosensitization of Bi2S3, thus exhibiting large and stable photocurrent response in the absence of electron sacrificial agent, that was 10.4 and 6.4 times higher than that of BiOI and Bi2S3, respectively. Then, a DBP PEC aptasensor was constructed by modifying the DBP aptamer on the surface of the ITO/Bi-BIS electrode. The aptasensor demonstrated a broad linear range (2-500 pM) and a low detection limit (0.184 pM). What's more, because there is no interference from electronic sacrificial agent, the aptasensor exhibited excellent selectivity in real water samples. Therefore, the proposed PEC has considerable potential for DBP monitoring.

Vanillic acid ameliorates diethyl phthalate and bisphenol S-induced oxidative stress and neuroinflammation in the hippocampus of experimental rats.

Long-term adverse effects on human health are caused by exogenous compounds that alter the functions of biological systems, especially neuroendocrine disruptors like diethyl phthalate (DEP) and bisphenol S (BPS). Although vanillic acid (VA) has pertinent neuropharmacological characteristics, its effect against DEP + BPS-induced neurotoxicity has not been explored. This study proposed that VA may offer protection against the neurotoxicity caused by DEP + BPS. Thirty male Wistar rats were randomly distributed across five groups: a control group receiving DMSO, a group exposed to a mixture of BPS and DEP, two BPS + DEP-exposed groups treated with VA at doses of 25 mg/kg or 50 mg/kg, and a nonexposed group treated with 50 mg VA/kg. After 21 days, the hippocampal tissues were processed for biochemical analyses. Our results indicate that exposure to DEP + BPS upregulated neurosignaling mediators (NTPDase, ADA, MAO-A, and Ca2+), inhibited others (AChE and Ca2+/Mg2+-ATPase), decreased hippocampus antioxidants (GSH, GPx, CAT, and SOD), and elevated markers of oxidative stress/damage (NO, H2O2, MDA, and AOPP). AR, BAX, TNF-α, BAK1, and IL-1ß expressions were upregulated, while IL-10 and BDNF expressions were downregulated. NF-κB and caspase-3/9 pathways were also upregulated. Co-treatment with vanillic acid remarkably precluded these neurotoxic outcomes by improving neurosignaling, augmenting antioxidant status, abrogating oxidative damage, inflammation (TNF-α, IL-1ß), and apoptosis (BAX, BAK1, caspase-3/9). Vanillic acid also restored IL-10 and BDNF levels, thereby exhibiting neuroprotective effects, corroborated by histological examinations. We posit vanillic acid as a safe and effective therapeutic agent against neurotoxicity occasioned by exposure to neuroendocrine disruptors.

Phthalate monoesters affect membrane fluidity and cell-cell contacts in endometrial stromal adherent cell lines and spheroids.

Phthalate monoesters have been identified as endocrine disruptors in a variety of models, yet understanding of their exact mechanisms of action and molecular targets in cells remains incomplete. Here, we set to determine whether epidemiologically relevant mono(2-ethyl-5-hydroxyhexyl) phthalate (MEHHP) can affect biological processes by altering cell plasma membrane fluidity or formation of cell-cell contacts. As a model system, we chose endometrial stromal cell lines, one of which was previously used in a transcriptomic study with MEHHP or MEHHP-containing mixtures. A short-term exposure (1 h) of membrane preparations to endocrine disruptors was sufficient to induce changes in membrane fluidity/rigidity, whereas different mixtures showed different effects at various depths of the bilayer. A longer exposure (96 h) affected the ability of cells to form spheroids and highlighted issues with membrane integrity in loosely assembled spheroids. Finally, in spheroids assembled from T-HESC cells, MEHHP interfered with the formation of cell-cell contacts as indicated by the immunostaining of zonula occludens 1 protein. Overall, this study emphasized the need to consider plasma membrane, membrane-bound organelles, and secretory vesicles as possible biological targets of endocrine disruptors and offered an explanation for a multitude of endocrine disruptor roles documented earlier.

[Content and Health Risks of Microplastics and Phthalate Esters in Bottled Water].

To study the content and health risks of microplastics （MPs） and phthalate esters （PAEs） in bottled water, a quantitative analysis of MPs was conducted by using Rose Bengal staining and stereomicroscopy. Seven PAEs were quantified by using gas chromatography-triple quadrupole tandem mass spectrometry （GC-MS/MS）. The daily intake of MPs was estimated and the carcinogenic and non-carcinogenic risks of PAEs were evaluated through a health risk assessment model. The results showed that the abundance of MPs in 21 bottled waters ranged from 48 n·L-1 to 216 n·L-1 （with the median abundance of 88 n·L-1）. The majority （72.1%） of MPs were fibrous in shape, and fragments accounted for only 27.9%. The average proportion of small-sized （10-50 µm） MPs was 33.9%, and that of large-sized MPs （>500 µm） was 4.3%. Most MPs were blue. The ∑(PAEs) in bottled water was 1.15-2.47 µg·L-1 （average 1.62 µg·L-1）. PAEs detected with high frequencies （100%） included dimethyl phthalate （DMP）, diethyl phthalate （DEP）, diisobutyl phthalate （DIBP）, di-n-butyl phthalate （DBP）, and di（2-ethylhexyl） phthalate （DEHP）, while the detection frequencies of butylbenzyl phthalate （BBP） and di-n-octyl phthalate （DNOP） were relatively low. The concentrations of DBP, DEHP, and DEP were all below the standard limits for drinking water in China. The ∑(PAEs) in the migration experiments was 0.61-2.04 µg·L-1 （average 1.33 µg·L-1）. The migration amounts of DBP and DEHP were also within the allowable range under the condition of 60℃ for 10 days. Seven PAEs were detected in both the bottles and caps, and the average content of DEHP in bottles was the highest, while DBP had the highest content in caps. The estimated intake of MPs （EDI） by drinking bottled water in different age groups of humans was 2.87 n·ï¼ˆkg·d）-1 for adults, 3.87 n·ï¼ˆkg·d）-1 for children, and 5.85 n·ï¼ˆkg·d）-1 for infants. The carcinogenic risks of DEHP in 21 bottled water samples and the migration test were less than the maximum acceptable risk level （1×10-6）, and the non-carcinogenic risk indices （HIs） of PAEs were all less than 1, indicating no non-carcinogenic risk to humans； however, the risk value of infants and children was higher than that of adults and should not be ignored.

Dual lateral flow test for simple and rapid simultaneous immunodetection of bisphenol A and dimethyl phthalate, two priority plastic related environmental contaminants.

Contamination of the environment by technogenic endocrine disrupting compounds (EDCs) becomes serious threat to public health. To effectively prevent this threat, it is necessary to improve analytical methods for EDCs to ensure mass, fast and productive monitoring. In the given work, a dual lateral flow test (LFT) is developed in the first time for simultaneous immunodetection of bisphenol A and dimethyl phthalate, priority EDCs releasing from plastic and belonging to different chemical classes. It combines integrated detection of two EDCs by one analytical system with rapidity and simplicity of LFTs allowing for off-lab testing without additional reagents and devices. Gold nanoparticles differing in shape and color (red gold nanospheres and blue gold nanoflowers) are applied as markers to simplify interpretation of the obtained results. Under the optimal conditions chosen for efficient control of the both analytes, the detection limits of bisphenol A and dimethyl phthalate are 0.67 ng/mL and 2.22 ng/mL, respectively. Time of the assay is 15 min. The proposed dual LFT has confirmed its practical applicability by analyzing natural water samples with recovery of bisphenol A and dimethyl phthalate in the ranges of 90.4-107.0% and 86.8-118.0%, respectively.

Phthalate Esters in Different Types of Cosmetic Products: A Five-Year Quality Control Survey.

Phthalate esters are commonly included in the formulations of cosmetics and related products in order to retain fragrance, enhance flexibility (i.e., by acting as plasticizers), facilitate the dissolution and dispersion of other ingredients, and improve the overall texture and sensory experience of the products. This study aimed to assess the presence and concentrations of phthalates in cosmetics by analyzing a comprehensive set of samples collected over a period of five years (2016-2020). The concentrations of nine different phthalate esters (BBP, DEHP, DNOP, DPP, DBP, DIPP, DMEP, DMP and PIPP) in 1110 cosmetics samples from France and Spain were determined by gas chromatography-mass spectrometry. The samples were included in five categories: soaps and shampoos; hand and body creams; lip gloss and lipsticks; nail polish; and facial makeup and skincare products. Some of the samples (4.86%) contained at least one phthalate at concentrations above the threshold limit (1 µg mL-1). Variable concentrations of different phthalates were determined in the 54 positive samples identified. DEHP was the most frequently detected phthalate, followed by DBP. The findings revealed different profiles according to the different categories of cosmetics and the phthalates detected in each. The results were critically compared with those obtained in various previous studies.

Di(2-ethylhexyl) phthalate exposure aggravates hypoxia/reoxygenation injury in cerebral endothelial cells by downregulating epithelial cadherin expression.

Di-(2-ethylhexyl) phthalate (DEHP) is a widely used plasticizer that has adverse health effects. Most phthalates exhibit reproductive toxicity and are associated with diseases such as cardiovascular disorders. However, the effect of DEHP exposure on acute hypoxia/reperfusion injury remains unknown. Therefore, we assessed whether hypoxia/reperfusion injury is aggravated by exposure to DEHP and investigated plausible underlying mechanisms, including oxidative stress and expression of cyclooxygenase-2 (COX-2)/prostaglandin E2 (PGE2) and endothelial junctional proteins. bEnd.3 cells were exposed to DEHP and subsequently subjected to oxygen-glucose deprivation (OGD). Cell viability was analyzed using 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) proliferation assay. The effect of DEHP/OGD/reoxygenation (R) was evaluated by assessing the levels of NO, reactive oxygen species (ROS), and PGE2. The expression of COX-2, cleaved caspase-3, cleaved PARP, inducible nitric oxide synthase (iNOS), and the endothelial tight junction proteins claudin-5 and ZO-1 was evaluated using quantitative polymerase chain reaction and western blotting. OGD/R decreased cell viability, and DEHP exposure before OGD/R further aggravated cell viability. DEHP/OGD/R significantly increased NO, PGE2, and ROS production following OGD/R. In the DEHP/OGD/R group, iNOS, COX-2, cleaved caspase-3, and cleaved PARP expression increased, and claudin-5 and ZO-1 levels decreased compared with those in the OGD/R group. E-Cadherin expression decreased significantly after DEHP/OGD/R exposure compared with that after OGD/R; this decrease in expression was recovered by treatment with the COX-2 inhibitor indomethacin and antioxidant N-acetylcysteine. Exposure to DEHP exacerbated hypoxia-reoxygenation injury. The enhanced damage upon DEHP exposure was associated with increased oxidative stress and COX-2 expression, leading to E-cadherin downregulation and increased apoptosis.

Responses of transcriptome and metabolome in peanut leaves to dibutyl phthalate during whole growth period.

Introduction: The application of agricultural film mulching technology has significantly contributed to increasing crop yield and income, but the pollution caused by residual film has seriously affected agricultural production and the natural environment. Agricultural film is commonly employed to enhance the yield of peanuts; its use may lead to excessive dibutyl phthalate (DBP) residues in peanut kernels. But, limited investigations have been conducted on the regulatory mechanism of peanut leaves in response to DBP exposure throughout the entire growth period. Methods: To bridge this knowledge gap, we investigated the differences in transcriptome and metabolome of peanut leaves under DBP stress. Results: According to visual observations, the results of morphological response showed that the growth of peanut plants was significantly inhibited from seedling to pod stage under DBP treatment. Transcriptomic analysis results showed that the genes AH19G05510 (LRR receptor-like serine threonine-protein kinase) and AH20G31870 (disease resistance), belonging to the FAR1 family and bZIP family respectively, may be key genes involved in the resistance to DBP stress throughout its growth stages. Metabolomic analysis results showed that during the initial stage of DBP stress, the key metabolites in peanut leaves response to stress were carboxylic acids and derivatives, as well as fatty acyls. As peanut growth progressed, flavonoids gradually became more prominent in the resistance to DBP stress. By integrating metabolomics and transcriptomics analysis, we have identified that purine metabolism during seedling and flowering stages, as well as the flavone and flavonol biosynthesis pathways during pod and maturity stages, played a crucial role in response to DBP stress. Discussion: These findings not only provide valuable key gene and metabolic information for studying anti-plasticizer pollution throughout the entire growth period of peanuts, but also offer reference for enhancing crop resistance to plasticizer pollution through genetic modification and metabolic regulation.

Revealing the bioavailability and phytotoxicity of different particle size microplastics on diethyl phthalate (DEP) in rye (Secale cereale L.).

Understanding how widely distributed microplastics (MPs) and diethyl phthalate (DEP) interact with crops remains limited, despite their significant implications for human exposure. We used physiology, transcriptomics, adsorption kinetics, and computational chemistry to assess rye's molecular response to two sizes of MPs (200 nm and 5 µm) and DEP, both individually and in combination. Findings systematically highlight potential ecological risks from MPs and DEP, with ecotoxicity ranking as follows: CK (Control Check) < LMPs < SMPs < DEP < LMPs+DEP < SMPs+DEP. Fluorescence and scanning electron microscopy revealed SMP's translocation ability in rye and its potential to disrupt leaf cells. DEP increased the electronegativity on MPs, which enhanced their uptake by rye. DEP adsorption by MPs in hydroponics reduced DEP bioavailability in rye (18.17-46.91 %). Molecular docking studies showed DEP interacted with chlorophyll, superoxide dismutase, and glutathione S-transferases proteins' active sites. Transcriptomic analysis identified significant up-regulation of genes linked to mitogen-activated protein kinase signaling, phytohormones, and antioxidant systems in rye exposed to MPs and DEP, correlating with physiological changes. These findings deepen the understanding of how MPs can accumulate and translocate within rye, and their adsorption to DEP raises crop safety issues of greater environmental risk.

Integrative models for environmental forecasting of phthalate migration from microplastics in aquaculture environments.

The pervasive utilization of plastic tools in aquaculture introduces significant volumes of microplastic fibers, presenting a consequential risk through the leaching of additives such as phthalates. This study scrutinizes the leaching dynamics of six prevalent phthalate esters (PAEs) from thirteen plastic aquaculture tools comprising polyethylene terephthalate (PET), polypropylene (PP), and polyethylene (PE), with ΣPAEs ranging from 0.24 to 4.26 mg g-1. Di(2-ethylhexyl) phthalate (DEHP) and dibutyl phthalate (DBP) emerged as predominant, marking significant environmental concern. Over a 30-day period, leaching quantities of Σ6PAEs from PET, PP, and PE fibers reached 36.65 µg g-1, 21.87 µg g-1 and 19.11 µg g-1, respectively, influenced by factors such as time, temperature, turbulence, and salinity. Notably, turbulence exerted the most pronounced effect, followed by temperature, with negligible influence from salinity. The kinetic models aligning with interface diffusion control was developed, predicting PAEs' leaching behavior with activation energies (Ea) indicative of the process's thermodynamic nature. The application of this model to real-world aquaculture waters forecasted significant risks, corroborating with empirical data and underscoring the pressing need for regulatory and mitigation strategies against PAEs contamination from aquaculture practices.

Associations between phenols, parabens, and phthalates and depressive symptoms: The role of inflammatory markers and bioinformatic insights.

Phenols, parabens, and phthalates are commonly found in consumer products, yet there is limited research on their individual and combined effects on depressive symptoms, particularly regarding the role of inflammation in these associations. This study aimed to evaluate these effects and explore potential molecular mechanisms, with a focus on inflammation as a mediator. We conducted a cross-sectional analysis involving 2766 adult participants from the National Health and Nutrition Examination Survey (NHANES) 2013-2016. Urine samples were analyzed for 15 chemicals, including 3 phenols, 2 parabens, and 10 phthalates. Depressive symptoms were assessed using the Patient Health Questionnaire-9 (PHQ-9). Statistical analyses included linear regression, restricted cubic splines, Bayesian Kernel Machine Regression and quantile g-computation models to investigate the relationships between chemical exposures and depressive symptoms. Additionally, mediation analysis was employed to explore the potential role of inflammation (immune cells, CRP, NLR) in these associations. The underlying molecular mechanisms were analyzed using bioinformatic approaches. Notably, BPA, MECPP, MEHHP, MiBP and MBP were found to be positively associated with depressive symptoms among females. Besides, BPA was the most significant positive contributor to the effect in the context of the chemical mixture, while the overall mixture effect was relatively weak. Furthermore, WBC were found to mediate a marginal portion (4 %) of the potential effects of MBP on depressive symptoms. The 15 genes identified are primarily involved in neurotransmission, mood regulation, and stress response. Further research is needed to elucidate the mechanisms underlying the observed associations.

Persistent effects of di-n-butyl phthalate on liver transcriptome: impaired energy and lipid metabolic pathways.

The environmental contaminant dibutyl phthalate (DBP) is reported to be hepatotoxic, but the underlying molecular pathways and pathological processes remain unclear. Here we used RNA-sequencing to characterize persistent hepatic transcriptional effects one week after the conclusion of five weeks oral exposure to 10 mg/kg/day or 100 mg/kg/day DBP in male mice. The exploratory transcriptome analysis demonstrated five differentially expressed genes (DEGs) in the 10 mg/kg/day group and thirteen in the 100 mg/kg/day group. Gene Set Enrichment Analysis (GSEA), which identifies affected biological pathways rather than focusing solely on individual genes, revealed nine significantly enriched Reactome pathways shared by both DBP treatment groups. Additionally, we found 54 upregulated and one downregulated Reactome pathways in the 10 mg/kg/day DBP group, and 29 upregulated and 13 downregulated pathways in the 100 mg/kg/day DBP group. According to the DEGs and the GSEA findings DBP exposure disrupts several key biological processes, including protein translation, protein folding, apoptosis, hedgehog signaling, degradation of extracellular matrix and alterations in the energy/lipid metabolism. Subsequent liver tissue analysis corroborated these findings, showing that DBP exposure induced tissue disorganization, oxidative stress, lipid accumulation, increased TNF-α, ATP and glucokinase levels. In addition, several proteins central for the metabolic system were affected, mostly in a dose-response pattern. Taken together the results show that DBP can cause hepatic stress and damage and suggest a potential role for DBP in the development of non-alcoholic fat liver disease, the most prevalent liver disease worldwide.

Urinary concentrations of phthalate and phthalate alternative metabolites and sperm DNA methylation: A multi-cohort and meta-analysis of men in preconception studies.

Phthalates are ubiquitous pollutants in the environment; however, the mechanisms of phthalate-associated reproductive disorders in men are not fully understood. The aim of this study is to investigate associations between urinary phthalate metabolite concentrations and sperm DNA methylation. The study was conducted on 697 men from three prospective pregnancy cohorts: Longitudinal Investigation of Fertility and the Environment (LIFE) Study, Sperm Environmental Epigenetics and Development Study (SEEDS), and Environment and Reproductive Health (EARTH) Study. Eighteen phthalate and two phthalate alternative metabolites were quantified by mass spectrometry in preconception urinary samples and sperm DNA methylation was measured via Illumina EPIC Array (v1). Regional methylation analyses were conducted to identify cohort-specific loci associated with urinary phthalate metabolites. Models were adjusted for age, body mass index (BMI), race, smoking status, urinary creatinine/specific gravity, and analytical batch for phthalate measurements. The cohort-specific results were meta-analyzed using METAL. Participants had an average age of 30 years, most (79.6 %) of whom had BMI>25 kg/m2 and were non-smokers (90.1 %). A total of 7,979 differentially methylated regions (DMRs; 7,979 LIFE-specific DMRs, 72 SEEDS-specific DMRs, and 23 EARTH-specific DMRs) were associated with urinary MBzP, MiBP, MMP, MCNP, MCPP, MBP, and MCOCH. Meta-analysis identified fewer DMRs than cohort-specific models: 946 DMRs were associated with MBzP, 27 DMRs associated with MiBP, and 1 DMR associated with MEHP. The majority of cohort-specific and meta-analysis-derived DMRs displayed a positive association with phthalate metabolite concentrations and were enriched in genes associated with spermatogenesis, response to hormones and their metabolism, embryonic organ development and developmental growth. In conclusion, several preconception urinary phthalate metabolites were associated with increased DNA methylation patterns in sperm. These findings provide an epigenetic pathway by which environmental phthalate exposures can impact couples' reproductive outcomes.

Occurrence, bioaccumulation, and partitioning of phthalate acid esters in the third largest freshwater lake (Lake Taihu) in China.

Phthalate acid esters (PAEs) are a category of plasticizers that are ubiquitous in freshwater environments attributable to extensive utilization. We collected water, suspended particulate matter (SPM), surface sediments, phytoplankton, and zooplankton from 23 sampling sites to investigate and complement the occurrence, bioaccumulation, and partitioning of five PAEs including dimethyl phthalate (DMP), diethyl phthalate (DEP), di-n-butyl phthalate (DBP), butyl benzyl phthalate (BBP), and di (2-ethylhexyl) phthalate (DEHP) in the third largest freshwater lake (Lake Taihu) of China. PAEs were extracted using Soxhlet extraction and solid phase extraction, and determined by gas chromatography-mass spectrometry. The average concentrations of the five PAEs in the water column, SPM, sediments, phytoplankton, and zooplankton of Lake Taihu were 1.93 ± 1.57 µg L-1, 765 ± 766 µg g-1, 1.68 ± 1.47 µg g-1, 1358 ± 1877 µg g-1, and 72.7 ± 134 µg g-1, respectively. DBP and DEHP were the dominant PAE congeners in the five environment compartments. The logarithmic concentrations of DBP, BBP, and DEHP in the SPM were negatively correlated with the logarithmic content of the SPM. Biodilution significantly impacted the occurrence of PAEs in the plankton. Bioaccumulation of PAEs was found in the plankton with log BCF (bioconcentration factor) in the phytoplankton ranging from 1.78 ± 0.86 to 4.13 ± 1.23 and log BAF (bioaccumulation factor) in the zooplankton varying from -0.10 ± 0.26 to 3.04 ± 0.64. Biomagnification of the PAEs from phytoplankton to zooplankton was not observed. DMP, DEP, and BBP migrated from sediments to water. DBP was in dynamic equilibrium in the sediment-water system. DEHP transferred from water to sediments. Our results provide crucial complementary knowledge on bioaccumulation and transfer of PAEs in planktonic food web, and their partitioning in different compartments of waters.

Exposure to phthalate plasticizer compromises normal brain function in an adult vertebrate.

Phthalates are key additives in many plastic products and among the most frequently used plasticizers. The release of some of them into the environment has been shown to have serious effects on development and reproduction. Based on such effects, diisononyl phthalate (DINP) has been advocated as a safer alternative to di-2-ethylhexyl phthalate (DEHP). Recently, it has been suggested that DEHP may affect the vertebrate blood-brain barrier. This could have serious consequences not only for the developing, but also for the adult brain. Here we tested for such impact on neuronal function and demonstrate acute exposure effects of both plasticizers on fundamental aspects of brain function in an adult vertebrate. We used the Mauthner neuron in the hindbrain of fish and its diverse inputs from various sensory systems as a model. After exposing intact goldfish to environmentally relevant plasticizer concentration (either 100 µg L-1, or 10 µg L-1), we show from in vivo intracellular recording that one month of environmental exposure to DEHP or DINP affected the sensory input to this central neuron, offset the balance between excitation and inhibition, and reduced its conduction speed by 20 %. The effects of both plasticizers were strong even at the concentration of 10 µg L-1. In an adult vertebrate, our findings thus demonstrate a previously neglected high sensitivity of various crucial brain functions to the acute exposure to phthalates.

Sulforaphane substantially impedes testicular ferroptosis in adult rats exposed to di-2-ethylhexyl phthalate through activation of NRF-2/SLC7A11/GPX-4 trajectory.

Di-2-ethylhexyl phthalate (DEHP) is a common plasticizer with a deleterious impact on testicular functionality and male fertility. Growing evidence implicates ferroptosis as one of the plausible mechanisms for DEHP-induced testicular injury. Sulforaphane (SFN) is a natural isothiocyanate displaying beneficial effects on testicular injury in several animal models. Herein, we explored the potential protective effect of SFN on testicular ferroptosis and toxicity evoked by DEHP. Adult male Wistar rats were equally distributed into three groups (n = 6/group): (i) CON group; (ii) DEHP group, received DEHP (2 g/kg PO) for 4 weeks; and (iii) DEHP + SFN group, received SFN (10 mg/kg, PO) 1 week prior to DEHP then concurrently with DEHP for further 4 weeks. Compared to CON group, exposure to DEHP caused testicular atrophy, deteriorated testicular architecture, testicular fibrosis, reduced sperm count and motility, higher sperm deformity, and declined serum testosterone level. All these abnormalities were ameliorated by SFN preconditioning. Additionally, pretreatment with SFN reversed the increased aromatase level and upregulated the steroidogenic markers in testes of DEHP-exposed rats. SFN pretreatment also counteracted DEHP-induced oxidative stress and boosted the total antioxidant capacity in testicular tissue via activation of the nuclear factor erythroid 2-related factor 2 (NRF-2) and its downstream target, hemeoxygenase-1 (HO-1). Moreover, SFN preconditioning mitigated DEHP-induced ferroptosis through up-surging SLC7A11, GPX-4, and GSH, while suppressing iron overload and ACSL4-induced lipid peroxidation in testicular tissue of rats. These findings may nominate SFN as a promising protective intervention to alleviate testicular ferroptosis associated with DEHP exposure through activation of NRF-2/SLC7A11/GPX-4 trajectory.

Phthalate exposure induces cell death and ferroptosis in neonatal microglial cells.

Background/aim: Phthalates are the materials used for plasticizing polyvinyl chloride. Di-(2-Ethylhexyl) phthalate (DEHP) is one of the phthalates most frequently used in a wide range of applications, including medical equipment such as endotracheal and feeding tubes, intravenous catheters, central lines, extracorporeal membrane oxygenation sets, total parenteral nutrition bags, blood product sets, and intravenous pump lines, respiratory sets in neonatal intensive care units (NICUs). Studies have shown that phthalates, including DEHP, can cross the placenta and blood-brain barrier, possibly leading to neurodevelopmental impairment in vitro and in vivo. However, the molecular mechanisms affected by phthalate exposure have not been explored in depth. This study aimed to illuminate the effects of DEHP on neuroinflammation at the molecular level using neonatal microglial cells as the model. Materials and methods: Mouse BV-2 neonatal microglia cells were exposed to DEHP under controlled conditions. Cellular toxicity was assessed via a cell viability assay and specific markers were used to evaluate the apoptosis/necrosis, cellular iron content, reactive oxygen species (ROS), and organelle integrity. Proinflammatory proteins were quantified using enzyme-linked immunosorbent assay, while ferroptosis was assessed using a ferroptosis blocker, and affected gene expressions were determined using quantitative reverse-transcriptase real-time polymerase chain reaction (RT-PCR). Results: The results revealed that high concentrations of DEHP exposure increased toxicity via increased levels of ROS and inflammation. Elevated ROS levels were observed to increase the tendency for mitochondrial-lysosomal disruption, bringing about apoptosis or necrosis. Moreover, iron homeostasis was dysregulated by DEHP, which putatively triggered ferroptosis in a dose-dependent manner. Conclusion: This study indicates that neonatal exposure to DEHP may be linked to neurodevelopmental impairment via inflammation-related cell death and ferroptosis. The prevalence of DEHP in NICU medical devices raises concerns about potential neurodevelopmental deficits, including disorders like autism and mental retardation. These findings highlight the urgency of addressing DEHP exposure in neonatal care.