Filling the Blank Space: Branched 4-Nonylphenol Isomers Are Responsible for Robust Constitutive Androstane Receptor (CAR) Activation by Nonylphenol.

4-Nonylphenol (4-NP), a para-substituted phenolic compound with a straight or branched carbon chain, is a ubiquitous environmental pollutant and food contaminant. 4-NP, particularly the branched form, has been identified as an endocrine disruptor (ED) with potent activities on estrogen receptors. Constitutive Androstane Receptor (CAR) is another crucial nuclear receptor that regulates hepatic lipid, glucose, and steroid metabolism and is involved in the ED mechanism of action. An NP mixture has been described as an extremely potent activator of both human and rodent CAR. However, detailed mechanistic aspects of CAR activation by 4-NP are enigmatic, and it is not known if 4-NP can directly interact with the CAR ligand binding domain (LBD). Here, we examined interactions of individual branched (22NP, 33NP, and 353NP) and linear 4-NPs with CAR variants using molecular dynamics (MD) simulations, cellular experiments with various CAR expression constructs, recombinant CAR LBD in a TR-FRET assay, or a differentiated HepaRG hepatocyte cellular model. Our results demonstrate that branched 4-NPs display more stable poses to activate both wild-type CAR1 and CAR3 variant LBDs in MD simulations. Consistently, branched 4-NPs activated CAR3 and CAR1 LBD more efficiently than linear 4-NP. Furthermore, in HepaRG cells, we observed that all 4-NPs upregulated CYP2B6 mRNA, a relevant hallmark for CAR activation. This is the first study to provide detailed insights into the direct interaction between individual 4-NPs and human CAR-LBD, as well as its dominant variant CAR3. The work could contribute to the safer use of individual 4-NPs in many areas of industry.

New insights into the regulation mechanism of Pacific white shrimp (Litopenaeus vannamei) hepatopancreas under 4-nonylphenol exposure using transcriptome analysis.

4-Nonylphenol (4-NP) is one of the common endocrine-disrupting chemicals (EDCs) in estuaries and coastal zones, which can exert detrimental effects on the physiological function of aquatic organisms. However, the molecular response triggered by 4-NP remains largely unknown in Pacific white shrimp (Litopenaeus vannamei). In this study, transcriptomic analysis was performed to investigate the underlying mechanisms of 4-NP toxicity in the hepatopancreas of L. vannamei. Nine RNA-Seq libraries were generated from L. vannamei at 0 h, 24 h, and 48 h following exposure to 4-NP. Compared with 0 h vs 24 h, 962 up- and 463 down-regulated differentially expressed genes (DEGs) were identified, indicating that many genes in L. vannamei were induced to resist adverse circumstances by 4-NP exposure. In contrast, 902 up- and 1027 down-regulated DEGs were revealed in the comparison of 0 h vs 48 h, demonstrating that prolonged exposure to the stress from 4-NP resulted in more inhibited genes. To validate the accuracy of the transcriptome data, eight DEGs were selected for quantitative real-time polymerase chain reaction (qRT-PCR), which were consistent with the RNA-Seq results. Through KEGG pathway enrichment analysis, three specific pathways related to hormonal effects and endocrine function of L. vannamei were enriched significantly, including tyrosine metabolism, insect hormone biosynthesis, and melanogenesis. After 4-NP stress, genes involved in tyrosine metabolism (Tyr) and melanogenesis pathway (AC, CBP, Wnt, Frizzled, Tcf, and Ras) were induced to promote melanin pigment to help shrimp resist adverse environments. In the insect hormone biosynthesis, ALDH, CYP15A1, CYP15A1/C1, and JHE genes were activated to synthesize juvenile hormone (JH), while Spook, Phm, Sad, and CYP18A1 were induced to generate molting hormone. There is an enhanced interaction between the molting hormone and JH, with JH playing a dominant role and maintaining its "classic status quo action". Our study demonstrated that 4-NP exposure led to impairments of biological functions in L. vannamei hepatopancreas. The genes and pathways identified provide novel insights into the molecular mechanisms underlying 4-NP toxicity effects in prawns and enrich the information on the toxicity mechanism of crustaceans in response to EDCs exposure.

A novel core-shell composite of PCN-222@MIPIL for ultrasensitive electrochemical sensing 4-nonylphenol.

A novel core-shell composite of PCN-222 and molecularly imprinted poly (ionic liquid) (PCN-222@MIPIL) with high conductivity and selectivity was prepared for electrochemical sensing 4-nonylphenol (4-NP). The electrical conductivities of some MOFs including PCN-222, ZIF-8, NH2-UIO-66, ZIF-67, and HKUST-1 were explored. The results indicated that PCN-222 had the highest conductivity and was then used as a novel imprinted support. PCN-222@MIPIL with core-shell and porous structure was synthesized using PCN-222 as support and 4-NP as template. The average pore volume of PCN-222@MIPIL was 0.085 m3 g-1. In addition, the average pore width of PCN-222@MIPIL was from 1.1 to 2.7 nm. The electrochemical response for PCN-222@MIPIL sensor for 4-NP was 2.54, 2.14, and 4.24 times that of non-molecularly imprinted poly (ionic liquid) (PCN-222@NIPIL), PCN-222, and MIPIL sensors, respectively, which result from superior conductivity and imprinted recognition sites of PCN-222@MIPIL. The current response of PCN-222@MIPIL sensor to 4-NP concentration from 1 × 10-4 to 10 µM presented an excellent linear relationship. The detection limit of 4-NP was 0.03 nM. The synergistic effect between the PCN-222 supporter with high conductivity, specific surface area and shell layer of surface MIPIL results in the outstanding performance of PCN-222@MIPIL. PCN-222@MIPIL sensor was adopted for detecting 4-NP in real samples and presented to be a reliable approach for determining 4-NP.

Exposure to 4-nonylphenol compromises Leydig cell development in pubertal male mice.

Exposure to 4-nonyl phenol (4-NP) on Leydig cell (LC) development and function remains poorly understood. We explored the effects of 4-NP on LC development and elucidate the underlying mechanisms. Male (28-day-old) mice received orally 4-NP (0.125, 0.25, and 0.5 mg/kg/day) for 28 days. We found that 4-NP at ≥ 0.125 mg/kg markedly compromised serum testosterone levels and LC numbers. Gene and protein expression analysis demonstrated downregulation of key genes and their proteins involved in LC steroidogenesis, including Star, Cyp11a1, Cyp17a1, Hsd17b3, Hsd3b6, and Scarb1. Furthermore, exposure to 4-NP induced oxidative stress, as evidenced by elevated reactive oxygen species (ROS) and malondialdehyde (MDA), as well as reduced superoxide dismutase 1/2 and catalase (CAT). Apoptosis was also observed in LCs following exposure to 4-NP, as shown by an increased BAX/BCL2 ratio and caspase-3. A TM3 mouse LC line further confirmed that 4-NP induced ROS and the expression of apoptosis-related genes and proteins. In conclusion, this study demonstrates that 4-NP exposure compromises LC development through multiple mechanisms.

Degradation of 4-nonylphenol in marine sediments using calcium peroxide activated by water hyacinth (Eichhornia crassipes)-derived biochar.

The contamination of marine sediments by 4-nonylphenol (4-NP) has become a global environmental problem, therefore there are necessaries searching appropriate and sustainable remediation methods for in-situ applications. Herein, water hyacinth [(WH) (Eichhornia crassipes)]-derived metal-free biochar (WHBC) prepared at 300-900 °C was used to promote the calcium peroxide (CP)-mediated remediation of 4-NP-contaminaed sediments. At [CP] = 4.37 × 10-4 M, [WHBC] = 1.5 g L-1, and pH = 6.0, the degradation of 4-NP was 77% in 12 h following the pseudo-first order rate law with rate constant (kobs) of 4.2 × 10-2 h-1. The efficient 4-NP degradation performance and reaction mechanisms of the WHBC/CP system was ascribed to the synergy between the reactive species (HO• and 1O2) at the WHBC surface on which there were abundant electron-rich carbonyl groups and defects/vacancies in the catalyst structure provides active sites, and the ability of the graphitized carbon framework to act as a medium for electron shuttling. According to microbial community analysis based on amplicon sequence variants, bacteria of the genus Solirubrobacter (Actinobacteria phylum) were dominant in WHBC/CP-treated sediments and were responsible for the biodegradation of 4-NP. The results showed great promise and novelty of the hydroxyl radical-driven carbon advanced oxidation processes (HR-CAOPs) that relies on the value-added utilization of water hyacinth for contaminated sediment remediation in achieving circular bioeconomy.

Effects of 4-nonylphenol on adipogenesis in 3T3-L1 preadipocytes and C3H/10T1/2 mesenchymal stem cells.

Obesogens are a subset of endocrine disruptor chemicals (EDCs) that cause obesity. The typical EDC 4-nonylphenol (4-NP) has been identified as an obesogen. However, the in vitro effects of 4-NP on adipogenesis remain unclear. In this study, 3T3-L1 preadipocytes and C3H/10T1/2 mesenchymal stem cells (MSCs) were used to investigate the influence of 4-NP on adipogenesis. The differentiation protocols for 3T3-L1 preadipocytes and C3H/10T1/2 MSCs took 8 and 12 days, respectively, beginning at Day 0. In differentiated 3T3-L1 preadipocytes, 20 µM 4-NP decreased cell viability on Days 4 and 8. Exposure to 4-NP inhibited triglyceride (TG) accumulation and adipogenic marker expression on Days 0-8, but the inhibitory effects were weaker on Days 2-8. The protein expression of pSTAT3 or STAT3 decreased on Days 0-8 and 2-8. Conversely, 4-NP promoted TG accumulation and the adipogenic marker expression in C3H/10T1/2 adipocytes. The opposing effects were attributed to physiological differences between the two cell lines. The 3T3-L1 preadipocytes are dependent on mitotic clonal expansion (MCE) to drive differentiation, while C3H/10T1/2MSCs and human preadipocytes are not. Additionally, 4-NP downregulated ß-catenin expression in C3H/10T1/2 adipocytes. Accordingly, we hypothesized that 4-NP promotes adipogenesis. The role of the canonical Wnt pathway in the promotion of adipogenesis by 4-NP requires further validation. This study provides new insights into the mechanisms and appropriate risk management of 4-NP.

Developmental immunotoxicity and its potential gender differences of perinatal exposure to 4-nonylphenol on offspring rats: JAK-STAT signaling pathway involved.

The aim of our study was to explore the developmental immunotoxicity (DIT) and its potential gender differences of perinatal exposure to 4-nonylphenol (4-NP), which was significant for the risk assessment of 4-NP exposure to fetuses and infants. Wistar pregnant rats were given the National Institution of Health (NIH)- 31 modified feed containing 0, 10, 100 and 500 mg/kg 4-NP from the gestation day (GD) 6 to the postnatal day (PND) 21. At PND21, the offspring rats were randomly selected to detect developmental immunotoxicity related indicators. Results suggested that high-dose 4-NP perinatal exposure caused growth retardation in infancy of male offspring rats, which was not obvious in female offspring rats. Also, 4-NP perinatal exposure induced DIT (mainly manifested as immunosuppression) with potential gender differences, including decreased weight of immune organs, suppressed immune function, decreased ratio of transforming growth factor (TGF)-ß/interleukin (IL)- 17A, increased ratio of T helper (Th) 17/regulatory T (Treg) cells et al. In addition, exploration of the Janus kinase (JAK)-signal transducer and activator of transcription (STAT) signaling pathway showed that JAK-STAT pathway mediated the leftward of Th17/Treg cells balance. Furthermore, the DIT to female offspring rats was more sensitive than to the males, which may be related to the differences of biological processes involved and needed to be further explored.

Multi- and transgenerational biochemical effects of low-dose exposure to bisphenol A and 4-nonylphenol on testicular interstitial (Leydig) cells.

Bisphenol A (BPA) and 4-nonylphenol (NP) are well-known endocrine-disrupting chemicals (EDCs) that have been proven to affect Leydig cell (LC) functions and testosterone production, but whether BPA and NP have multi- and transgenerational biochemical effects on Leydig cells (LCs) is unknown. Fourier transform infrared (FTIR) spectroscopy is a powerful analytical technique that enables label-free and non-destructive analysis of the tissue specimen. Herein we employed FTIR coupled with chemometrics analysis to identify biomolecular changes in testicular interstitial (Leydig) cells of rats after chronic exposure to low doses of BPA and NP. Cluster segregations between exposed and control groups were observed based on the fingerprint region of 1800-900 cm-1 in all generations. The main biochemical alterations for segregation were amide I, amide II and nucleic acids. BPA and NP single and co-exposure induced significant differences in the ratio of amide I to amide II compared to the corresponding control group in all generations. BPA exposure resulted in remarkable changes of cellular gene transcription and DNA oxidative damage across all generations. Direct exposure to BPA, NP, and BPA&NP of F0 and F1 generations could significantly decrease lipid accumulation in LCs in the F2 and F3 generations. The overall findings revealed that single or co-exposure to BPA and NP at environmental concentrations affects the biochemical structures and properties of LCs.

[4-nonylphenol combined with bisphenol A induces testicular autophagy and spermatogenic impairment by inhibiting the Akt/mTOR pathway in F1 male offspring rats].

OBJECTIVE: To study the impacts of exposure of F0 generation rats to 4-nonylphenol (4-NP) and bisphenal A (BPA) on the autophagy of testicular cells and key gene expressions of the Akt/mTOR pathway in the F1 generation offspring rats. METHODS: Using a 2×2 factorial design, F0 female and male rats were randomly assigned to receive intragastrically sigma vegetable oil (the control), BPA at 0.5 mg/kg, 4-NP at 5 mg/kg, and BPA+4-NP both at 0.5 mg/kg, respectively, qd alt for 30 days. Then the rats in each group were mated at a ratio of 1∶3. After pregnancy, the female rats continued the above intragastrical administration till delivery. The F1 generation male offspring rats were killed at 60 postnatal days, and their testes harvested for sperm count, observation of the morphological and autophagic changes of the testis, and determination of the relative mRNA expression levels of Akt, mTOR, 4EBP1 and p70S6K and protein expression levels of LC3-I and LC3-II. RESULTS: The sperm count of the F1 generation male offspring rats was markedly decreased in the BPA and BPA+4-NP groups compared with that in the control (P < 0.05) but no significant interactive effect was observed between BPA and 4-NP (P > 0.05). The ratio of LC3-II / LC3-I was remarkably increased in the 4-NP, BPA and BPA+4-NP groups in comparison with that in the control (P < 0.05), and a significant interactive effect was shown between BPA and 4-NP (P < 0.05). The cells in the seminiferous tubules of the rats in the 4-NP, BPA and BPA+4-NP groups were loosely arranged, with a decreased count of sperm and increased number of autophagic vacuoles. Significant down-regulation was observed in the relative mRNA expression of p70S6K in the 4-NP group, as well as in that of mTOR in the BPA group and Akt in the BPA+4-NP group (all P < 0.05). A remarkably up-regulated expression of 4EBP1 mRNA was found in all the three intervention groups (P < 0.05), with a significant interactive effect between BPA and 4-NP on the expressions of Akt and 4EBP1 mRNA (P < 0.05). CONCLUSION: Long-term exposure to low-concentration BPA and 4-NP impairs the testicular function of the F1 generation male offspring rats, which is closely related to the autophagy of testicular cells and changes of the Akt/mTOR pathway.

The mechanism of cytotoxicity of 4-nonylphenol in a human hepatic cell line involves ER-stress, apoptosis, and mitochondrial dysfunction.

4-Nonylphenol (4-NP) is an emerging environmental pollutant widely diffused in waters and sediments. It mainly derives from the degradation of alkyl phenol ethoxylates, compounds commonly employed as industrial surfactants. 4-NP strongly contaminates foods and waters for human use; thus, it displays a wide range of toxic effects not only for aquatic organisms but also for mammals and humans. After ingestion through the diet, it tends to accumulate in body fluids and tissues. One of the main organs where 4-NP and its metabolites are concentrated is the liver, where it causes, even at low doses, oxidative stress and apoptosis. In the present study, we analyzed the effects of 4-NP on a human hepatic cell line (HepG2) to deepen the knowledge of its cytotoxic mechanism. We found that 4-NP, in a range of concentration from 50 to 100 µM, significantly reduced cell viability; it caused a partial block of proliferation and induced apoptosis with activation of caspase-3 and overexpression of p53. Moreover, 4-NP induced-apoptosis seemed to involve both an ER-stress response, with the appearance of high level of GRP78, CHOP and the spliced XBP1, and a dysregulation of mitochondrial physiology, characterized by an overexpression of main markers of mitochondrial dynamics. Our data support the idea that a daily consumption of 4-NP-contaminated foods may lead to local damages at the level of gastrointestinal system, including liver, with negative consequences for the organ physiology.

Exposure to 4-nonylphenol induces a shift in the gene expression of gsdf and testis-ova formation and sex reversal in Japanese medaka (Oryzias latipes).

The branched isomer mixture 4-nonylphenol (4-NP) has been used worldwide as a surfactant, and can have endocrine-disrupting effects on aquatic organisms. For instance, 4-NP induces the formation of testis-ova (i.e., testicular and ovarian tissue in the same gonad) or male to female sex reversal of various teleost fishes. Recently, our group revealed that altered gsdf gene expression is associated with disruption of gonadal differentiation in Japanese medaka (Oryzias latipes) embryos exposed to methyltestosterone or bisphenol A, suggesting that gsdf might be useful as a biomarker for predicting the impact of endocrine-disrupting chemicals (EDCs) on gonadal differentiation. Here, we used 4-NP to examine further whether gsdf expression at the embryo stage is useful for predicting EDC impact on gonadal sex differentiation. When fertilized medaka eggs were exposed to 32 or 100 µg/L 4-NP, testis-ova in genetic males and sex reversal from genetic male to phenotypic female were observed. At stage 38 (just before hatching), 4-NP exposure at 1-100 µg/L did not affect gsdf expression in XX embryos compared with the nontreated control; however, in XY embryos, the gsdf expression in the 100 µg/L-exposed group was significantly lower than that in the controls. The 4-NP concentration at which gsdf expression was suppressed was equal to that at which testis-ova and sex reversal were induced. These results indicate that expression of the gsdf gene at the embryonic stage in medaka is a useful biomarker for predicting the impact of EDCs on sexual differentiation.

A prominent environmental endocrine disruptor, 4-nonylphenol, promotes endometriosis development via plasmacytoid dendritic cells.

Endometriosis is an estrogen-dependent chronic inflammatory disease and is associated etiologically with environmental endocrine disruptor (EED) exposure. 4-nonylphenol (NP), a widely found EED, has weak estrogenic activity and modulates plasmacytoid dendritic cell (pDC) function in vitro and in vivo. We aimed to elucidate the immunomodulatory effect of NP on the development of endometriosis, particularly focusing on pDCs. This study established a surgically induced endometriosis murine model (C57BL/6) under conditions of NP treatment that are relevant to the level and route of human exposure. Multi-parametric flow cytometry was used for analysis of infiltrated immune cell subsets in lesions. The results showed that NP exposure significantly promoted endometriotic lesion growth, survival and angiogenesis development of lesions as well as pDC accumulation in the lesions in mice. Adoptive transfer of NP-conditioned pDCs into mice significantly enhanced lesion development and local pDC infiltration, whereas NP-conditioned conventional dendritic cells did not affect lesion growth. In vitro functional analysis showed that NP-conditioned pDCs in lesions expressed high levels of CD36, a scavenger receptor and NP-conditioned splenic pDCs secreted an enhanced level of IL-10 in response to apoptotic cell recognition in a CD36-dependent manner. Furthermore, we observed that local treatment with blocking antibodies against IL-10 and CD36 on the day of surgery significantly inhibited lesion development. NP exposure also altered the estrous cycle in mice. The results suggest that chronic and low-dose exposure to NP enhances endometriotic lesion growth by altering pDC homeostasis and function. This study has important implications for understanding the environment-innate immunity interaction in human endometriosis.

Removal and biotransformation of 4-nonylphenol by Arthrospira maxima and Chlorella vulgaris consortium.

4-Nonylphenol (4-NP) is an anthropogenic contaminant found in different environmental matrices that has an effect over the biotic and abiotic factors within the environment. Bioremediation by microorganisms can be used as a potential treatment to remove this pollutant. In this work, a consortium of two microorganisms, Arthrospira maxima and Chlorella vulgaris, was employed to remove 4-NP from water. The parameters analyzed included cell growth, removal of 4-NP, and 4-NP remnant in the biomass. In addition, the metabolites produced in the process by this consortium were identified. It was found that C. vulgaris is more resistant to 4-NP than A. maxima (cell growth inhibition by 4-NP of 99%). The consortium used in this study had an IC50 greater than any strain of microalgae or cyanobacteria reported for 4-NP removal (9.29 mg/L) and reduced up to 96% of 4-NP in water in the first 48 h of culture. It was also observed that there is a bio-transformation of 4-NP, comparable with the process carried out by another bacterium, in which three similar metabolites were found (4-(1-methyl-octyl)-4-hydroxy-cyclohex-2-enone, 4-nonyl-4-hydroxy-ciclohexa-2,5-dienone and 4-nonyl-4-hydroxy- ciclohex-2-enone) and one that is similar to plant metabolism (4-nonyl-(1-methyl,6,8-metoxy)-hydroxybenzene). These results indicate that microalgae and cyanobacteria consortium can be used to remove 4-NP from water.

An updated weight of evidence approach for deriving a health-based guidance value for 4-nonylphenol.

4-Nonylphenol (NP) is a persistent estrogen-active compound. Human exposure to NP is primarily through water and food. Although risk assessments of NP have been conducted by the European Union and a few other countries, only the Danish Veterinary and Food Administration, in 2000, proposed a tolerable daily intake of 0.005 mg kg-1 body weight (bw) day-1 . New data have been accumulated since then, prompting an update on the risk assessment of NP. A weight of evidence approach is recommended for use in scientific assessments by several agencies, e.g., European Food Safety Authority, etc. Based on the results of a weight of evidence approach, two methods were used to derive the health-based guidance value (HBGV) for NP in this study, namely a no observed adverse effects level/lowest observable adverse effect level method, and a benchmark dose method. Considering the considerable uncertainty of benchmark dose model fitting of the available data, a tolerable daily intake value of 0.025 mg kg-1 bw day-1 was derived as a provisional HBGV for NP based on the lowest observable adverse effect level value of 15 mg kg-1 bw day-1 of the renal toxicity in rats, together with the uncertainty factor of 600. However, the HBGV of NP still needs further investigation.

Evaluation of 4-nonylphenol and bisphenol A toxicity using multiple molecular biomarkers in the water flea Daphnia magna.

Alkylphenols are well-known endocrine disruptors and may cause developmental and reproductive disorders in aquatic organisms. Daphnia magna is commonly used in ecotoxicological studies as a promising model species to investigate the effects of endocrine distruptors. In the present study, transcriptional modulation of eleven potential molecular indicators related to detoxification, antioxidant, development, and cellular stress was analyzed in D. magna exposed to different concentrations of bisphenol A (BPA) and 4-nonylphenol (4-NP) for 24 h and 48 h, using real-time qPCR. A hierarchical clustering analysis was applied to investigate relations among molecular markers depending on the compound, exposure duration, and concentration. Our findings suggested that GSH-related systems and stress proteins may be involved in cellular defense against BPA and 4-NP-mediated toxicity with different modes of action. Furthermore, these compounds may interrupt molting and reproduction in daphnids. In particular, D. magna GSH-related genes seem to be strongly affected by 4-NP exposure, indicating their potential as molecular biomarkers.

[Risk of pathoosophermia in men with joint exposure to endocrine disruptors].

The purpose of the study is to assess the risk of pathozoospermia with the combined effect of bisphenol A, triclosan and 4-nonylphenol. MATERIALS AND METHODS: 84 samples of sperm were studied in men with normo-and patozoospermia. In seminal fluid, the concentrations of bisphenol A, triclosan and 4-nonylphenol were simultaneously determined by gas chromatography with mass spectrometry (GC-MS). The spermiological study was carried out according to the WHO recommendations (2010) taking into account the assessment of the number of spermatozoa, their motility and morphology, as well as sperm damage. The results were subjected to statistical processing using the Mann-Whitney U-test and the multiple logistic regression method. Results were considered statistically significant at p<0.05. RESULTS: Bisphenol A was found in 100% of the ejaculate samples with a median concentration of 0.150 ng/ml. Triclosan and 4-nonylphenol were detected in 84.3% and 98.1% of ejaculate samples with a median concentration of 0.11 ng/ml. and 0.16 ng/ml, respectively. The comparison groups were statistically significantly distinguished by the concentrations of bisphenol A and triclosan (p<0.001; p=0.018), respectively, as well as the DNA index of spermatozoa fragmentation (p=0.002). Bisphenol A, by increasing the concentration in seminal fluid by 0,1 ng/ml, increased the chance of developing pathosospermia by 24.9 times. The effect of triclosan and 4-nonylphenol on pathosospermia was not statistically significant. In a multiple logistic regression equation describing the joint effect of EDs on patozoospermia, as well as the ratio of the chances of its occurrence with a change in the concentration of one of the endocrine disruptors per unit while fixing the other variables, the coefficients of triclosan and 4-nonylphenol on the patozoospermia were also not statistically significant. In addition, the confidence interval for the odds ratio included one. CONCLUSION: The development of pathozoospermia is primarily associated with the effect of bisphenolA in seminal fluid in men. It should be assumed that there is no synergistic effect of endocrine disruptors on patozoospermia.

Fabrication of a highly sensitive electrochemical sensor based on electropolymerized molecularly imprinted polymer hybrid nanocomposites for the determination of 4-nonylphenol in packaged milk samples.

Herein, an imprinted electrochemical sensor based on graphene-Au nanoparticles incorporated with molecularly imprinted polymer (MIP) was fabricated for determination of 4-nonylphenol (4-NP). Grafted MIP electropolymerized on nanoscale multilayer films electrode was achieved using 4-NP as a template and P-aminothiophenol as a functional monomer. The electrochemical properties of the MIP nanoscale multilayer membrances were characterized and measured by cyclic voltammetry and differential pulse voltammetry techniques; using ferrocyanide/ferricyanide-redox marker. Several important parameters were optimized and investigated to improve the performance of the sensor. Under the optimized conditions, the developed sensor showed an excellent linear response over the concentration ranges of 50-500 ng mL-1 (4-NP) with a detection limit of 0.01 ng mL-1(S/N = 3). The developed sensor showed a good selective recognition of 4-NP compared with structural analogue, exhibited a good reproducibility and accuracy with long-term stability. At last, the feasibility of the proposed methodology was successfully applied fordetection of 4-NP in milk and its packaging materials.

Modulatory effects of green tea extract against the hepatotoxic effects of 4-nonylphenol in catfish (Clarias gariepinus).

The antioxidant role of the green tea (Camellia sinensis) extract (GTE) was examined to remedy the toxic effects of (0.2mgl-1) 4-nonylphenol(4-NP). Biochemical parameters, antioxidant enzymes, liver lipid peroxidation (LPO), DNA fragmentation, and apoptosis as well as histopathology of liver of African catfish Clarias gariepinus were considered. Catfishes were divided into four groups: first group (control), second group (0.2mgl-1 of 4-NP), third group (0.2mgl-1of 4-NP +100mg GTE l-1water), and fourth group (0.2mgl-1 of 4-NP +200mg GTE l-1water). The results showed that significant increments of serum glucose, AST, ALT, total protein, total lipids, cholesterol, G6PDH, and cortisol. Meanwhile, serum acetylcholinesterase, ALP, and LDH were significantly reduced. In addition, antioxidant enzymes (SOD, CAT, GST, and TAC) levels were reduced in 4-NP treated fish compared to control. Also, there were significant increments in hepatic LPO, DNA fragmentation, and apoptotic erythrocytes in 4-NP treated fish compared to control. Liver of 4-NP treated fish showed some histopathological alterations such as, vacuolization in hepatocytes, congestion in central vein, infiltration of mononuclear inflammatory cells, and necrosis as well as depletion of glycogen content of liver. Addition of green tea extract into the water restored the alterations in most of those biomarkers induced by 4-NP. We concluded that, GTE has a protective role against hepatic failure, depletion of antioxidant defense, and genotoxicity induced 4-NP in C. gariepinus.

Estrogenic chemicals at body burden levels attenuate energy metabolism in 3T3-L1 adipocytes.

The study aimed to examine effects of environmental estrogens at body burden levels on energy metabolism in fat cells. Acclimation of T47D-KBluc cells in estrogen-deprived medium was established for high performance of estrogen-responsive luciferase reporter assay. With the assay, relative estrogenic potency of four selected estrogen receptor (ER) agonists, i.e. diethylstilbestrol, ß-estradiol, 4-nonylphenol and bisphenol A, were determined. Immunoblot analysis revealed that the ER agonists at both EC80 and EC100 caused rapid and transient phosphorylation of extracellular signal-regulated kinases (ERK) in an ER-dependent manner. 3T3-L1 adipocytes treated with the ER agonists at EC80 for 24 hours exhibited significant downregulation in mitochondrial respiration and glycolytic function. Importantly, EC80 values of 4-nonylphenol (6.0 × 10-10  m) and bisphenol A (1.0 × 10-8  m) are in the range of human body burdens. The finding that estrogenic chemicals at body burden levels cause significant impact on fat cell energy metabolism raises an important public health issue that deserves more attention.

Induction of apoptosis and DNA damage by 4-nonylphenol in African catfish (Clarias gariepinus) and the antioxidant role of Cydonia oblonga.

In this study, we assessed the toxic effects of sub lethal concentration (0.1mgl-1) 4-nonylphenol (4-NP) on serum biochemical parameters, liver lipid peroxidation (LPO) and antioxidant enzymes of the African catfish Clarias gariepinus for 14 days and the ability of the quince leaf extract to alleviate the effects of (4-NP). Fish were categorized into four groups: control, exposure to 0.1mgl-1 4-NP, exposure to 0.1mgl-1 4-NP with quince leaf extract (10ml/30L water), and exposure to 0.1mgl-1 4-NP with quince leaf extract (20ml/30L water). 4-NP exposure induced a significant (p<0.05) increase in the levels of glucose, AST, ALT, creatinine, urea, uric acid, cholesterol, and G6PDH as well as, the percentages of hepatic LPO level, DNA fragmentation, and apoptotic erythrocytes (p<0.05). A significant (p<0.05) decrease in alkaline phosphatase (ALP), total protein, albumin, globulin, total lipids, and LDH were also recorded. Liver enzyme activities (SOD, CAT and TAC) were increased. Addition of the quince leaf extract into the water was able to reinstate the alterations in biochemical parameters, antioxidant biomarkers, apoptotic level and hepatic DNA damage induced by 4-NP.