Suspension array for multiplex immunoassay of five common endocrine disrupter chemicals.

A low cost and effective indirect competitive method is reported to detect five EDCs, 17-beta-estradiol (E2), estriol (E3), bisphenol A (BPA), diethylstilbestrol (DES), and nonylphenol (NP) simultaneously, based on suspension array technology (SAT). Five kinds of complete antigens (E2-BSA, E3-BSA, BPA-BSA, DES-BPA, NP-BSA) were coupled to different encoding microspheres using purpose-made solutions in our laboratory instead of commercially available amino coupling kits; the method was further optimized for determination and reducing  the cost. Encoding and signaling fluorescence of the particles are determined at 635/532 nm emission wavelengths. High-throughput curves of five EDCs were draw and the limit of detection (LOD) were between 0.0010 ng mL-1 ~ 0.0070 ng mL-1. Compared with traditional ELISA methods, the SAT exhibited better specificity and sensitivity. Experiments using spiked milk and tap water samples were also carried out, and the recovery was between 85 and 110%; the results also confirmed good repeatability and reproducibility. It illustrated great potential of the present strategy in the detection of EDCs in actual samples.

Endocrine-Disrupting Chemicals and Infectious Diseases: From Endocrine Disruption to Immunosuppression.

Endocrine-disrupting chemicals (EDCs) are hormonally active compounds in the environment that interfere with the body's endocrine system and consequently produce adverse health effects. Despite persistent public health concerns, EDCs remain important components of common consumer products, thus representing ubiquitous contaminants to humans. While scientific evidence confirmed their contribution to the severity of Influenza A virus (H1N1) in the animal model, their roles in susceptibility and clinical outcome of the coronavirus disease (COVID-19) cannot be underestimated. Since its emergence in late 2019, clinical reports on COVID-19 have confirmed that severe disease and death occur in persons aged ≥65 years and those with underlying comorbidities. Major comorbidities of COVID-19 include diabetes, obesity, cardiovascular disease, hypertension, cancer, and kidney and liver diseases. Meanwhile, long-term exposure to EDCs contributes significantly to the onset and progression of these comorbid diseases. Besides, EDCs play vital roles in the disruption of the body's immune system. Here, we review the recent literature on the roles of EDCs in comorbidities contributing to COVID-19 mortality, impacts of EDCs on the immune system, and recent articles linking EDCs to COVID-19 risks. We also recommend methodologies that could be adopted to comprehensively study the role of EDCs in COVID-19 risk.

A novel method for rapid and quantitative detection of bisphenol A in urine.

Bisphenol A (BPA) is classified as an endocrine disruptor (ED) and it can interact with variety of hormone receptors leading to hormonal disruption and increased risk of various adverse health effects. Reducing human exposure to BPA is one of the main challenges of public health, as it is constantly present in daily life. A low-cost and commonly applied method to enable determination of BPA in the patient's body has yet to be developed. Currently available techniques are expensive, time-consuming, and require access to highly equipped analytical chemistry laboratories. Here we describe a fast and cheap engineered lateral flow assay of our design, to detect of BPA in urine samples. The technology not only provides an opportunity to perform rapid medical diagnostics without the need for an access to the central laboratory but also a means for self-diagnosis by the patient. The addition of ß-glucuronidase improves the sensitivity of detection as it releases the free BPA from glucuronide complexes in urine. This invention may become a demonstrated analytical means for lowering human exposure to BPA and probably also to other EDs and consequently, may be useful in decrease of the risk for several lifestyle diseases.

Establishment of an immunofiltration strip for the detection of 17ß-estradiol based on the photothermal effect of black phosphorescence.

Recently, the photothermal effect of nanomaterials has opened the door for new appealing strategy, which can generate a promising and powerful tool when combined with immunoassay. As a new kind of nanomaterial, black phosphorus (BP) has aroused widespread interest. In this study, a novel immunofiltration strip method with temperature as the readout signal based on the photothermal effect of BP nanosheets was established. The temperature was monitored by a portable temperature sensor. Using an indirect competitive strategy, it provides a simple, rapid, sensitive, and economic platform for the detection of 17ß-estradiol, a kind of endocrine disrupting compound that is frequently detected in environmental water or food samples. The higher the concentration of 17ß-estradiol in the sample, the less BP nanosheets are brought to bind to the strip surface, along with lower temperature variation when exposed to intensive laser irradiation. Under optimum conditions, a detection limit of 0.104 ng mL-1 was achieved. The feasibility of this assay was assessed by a standard addition method in water and milk samples, showing good performance and indicating potential application value for easy-to-use, inexpensive, and on-site monitoring of 17ß-estradiol.

Sex-associated protective effect of early bisphenol-A exposure during enteric infection with Trichinella spiralis in mice.

Bisphenol A (BPA) is an endocrine disruptor compound with estrogenic activity, possessing affinity for both nuclear (ERα and ERß) and membrane estrogen receptors. The main source of BPA exposure comes from the contamination of food and water by plastic storage containers or disposable bottles, among others, in which case BPA is easily ingested. Exposure to BPA during early pregnancy leads to lifelong effects; however, its effect on the immune system has not been fully studied. Since endocrine and immune systems interact in a bidirectional manner, the disruption of the former may cause permanent alterations of the latter, thus affecting a future anti-parasitic response. In this study, neonate BALB/c mice were exposed to a single dose of BPA (250 µg/kg); once sexual maturity was reached, they were orally infected with Trichinella spiralis (T. spiralis). The analyses performed after 5 days of infection revealed a decreased parasitic load in the duodenum of mice in the BPA-treated group. Flow cytometry analyses also revealed changes in the immune cell subpopulations of the infected animals when compared to the BPA-treated group. RT-PCR analyses of duodenum samples showed an increased expression of TNF-α, IFN-γ, IL-4, IL-5, and IL-9 in the BPA-treated group. These findings show a new aspect whereby early-life exposure to BPA contributes to the protection against T. spiralis by modulating the anti-parasitic immune response.

Neutrophils life under estrogenic and xenoestrogenic control.

Over 100 years ago, scientists had identified cells that represent the most abundant population of peripheral blood leukocytes; they called this population neutrophils. Day by day, the knowledge specific to neutrophils is augmented with new and often surprising aspects and facts about neutrophils' life or death. Estrogens (estrone, estriol, and estradiol) are relevant for the regulation of immune responses that are related with neutrophils. An understanding of the molecular mechanism of the action of endogenous hormones allows us to predict the effects of the substances that commonly occur in an environment with estrogen-like properties (xenoestrogens (e.g., bisphenol A, DDT, tributyltin, polychlorinated biphenyls, nonylphenol and octylphenol)). Therefore, we summarize current literature on the impact of estrogens and xenoestrogens, on each aspect of neutrophil life, as well as describe its mechanism of actions in neutrophils.

A survey of 17α-ethinylestradiol and mestranol residues in Hawkesbury River, Australia, using a highly specific enzyme-linked immunosorbent assay (ELISA) demonstrates the levels of potential biological significance.

This study reports on the potential status of 17α-ethinylestradiol (EE2) and mestranol (MeEE2) residues in aquatic environments in New South Wales (NSW), Australia, based on the analysis by a specific ELISA we developed. Polyclonal antibodies were raised against the EE2 hapten with a linker attached at the C3-position to direct the antibody binding towards the ring D of EE2/MeEE2. Using this approach, an ELISA highly specific to EE2 and MeEE2 was successfully developed, showing less than 3.1% cross-reactivity (% CR) with other major steroidal sex hormones and their derivatives. The assay performed with the limit of detection (LOD) of 0.04 ± 0.01µg/L for both EE2 and MeEE2, and the limit of quantitation (LOQ) of 0.05 ± 0.01ng/L when it was coupled with the SM2-Biobeads solid phase extraction. Prior to conducting the survey study, it was validated against the gas chromatography-mass spectrophotometry (GC-MS) method, which showed high correlation with R2 of 0.934. Fresh surface water samples collected at different sites along Hawkesbury River in New South Wales (NSW) were analyzed for the EE2/ MeEE2 residues using the developed ELISA. The EE2/MeEE2 levels were found to range between 4.1 and 8.3ng/L in Emigrant Creek, NSW, where the primary activity was macadamia plantation, and higher levels between 15 and 29ng/L in South Creek, NSW, Greater Western Sydney at sites upstream and downstream of the municipal sewage treatment plants.

Environmental alkylphenols modulate cytokine expression in plasmacytoid dendritic cells.

BACKGROUND: Alkylphenols, such as nonylphenol (NP) and 4-octylphenol (4-OP), have the potential to disturb immune system due to their weak estrogen-like activity, an effect with potential serious public health impact due to the worldwide distribution of these substances. Plasmacytoid dendritic cells (PDCs) can secrete large amounts of type I IFNs and are critical in immune regulation. However, there has been limited study about the influence of alkylphenols on the function of pDCs. OBJECTIVE: The aim of this study was to examine the effect of alkylphenols on pDC functions in vitro and in vivo and then further explored the involved signaling pathways and epigenetic changes. METHODS: Circulating pDCs from human peripheral blood mononuclear cells were treated with alkylphenols with or without CpG stimulation. Alkylphenol-associated cytokine responses, signaling events, histone modifications and viral activity were further examined. In NP-exposed mice, the effect of NP on splenic pDC function and allergic lung inflammation were also assessed. RESULTS: The results showed that NP increased the expression of TNF-α, but suppressed IL-10 production in the range of physiological doses, concomitant with activation of the MKK3/6-p38 signaling pathway and enhanced levels of acetylated histone 3 as well as histone 4 at the TNFA gene locus. Further, in CpG-stimulated pDCs, NP suppressed type I IFNs production, associated with down-regulation of IRF-7 and MKK1/2-ERK-Elk-1 pathways and led to the impaired anti-enterovirus 71 activity in vitro. Additionally, splenic pDCs from NP-exposed mice showed similar cytokine changes upon CpG stimulation under conditions relevant to route and level of exposure in humans. NP treatment also enhanced allergic lung inflammation in vivo. CONCLUSION: Alkylphenols may influence pDCs' functions via their abilities to induce expression of a pro-inflammatory cytokine, TNF-α, and to suppress regulatory cytokines, including IL-10, IFN-α and IFN-ß, suggesting the potential impact of endocrine disrupting chemicals on immune regulation.

Facile and rapid magnetic relaxation switch immunosensor for endocrine-disrupting chemicals.

In order to develop facile, fast and sensitive detection methods for endocrine-disrupting chemicals (EDCs), we described a sensitive biosensing system involving magnetic relaxation switch, based on the assembly of cross-linked superparamagnetic iron oxide (CLIO) nanoparticles induced by the antigen-antibody biorecognition. The design of smart CLIO-based superparamagnetic iron oxide nanoparticles and antigen-OVA was described for the detection of bisphenol A [2,2-bis (4-hydroxyphenol) propane (BPA)]. The addition of BPA to the rapid magnetic relaxation switch immunosensor led to transverse relaxation time (T2) shortening compared to a blank control as shown by NMR relaxometry measurements. This process was also applied to the rapid and facile determination of concentrations of BPA in drinking water (tap water). Good linearity for all calibration curves was obtained, and the limit of detection (LOD) for BPA was 0.4 ng/mL in tap water.

Ontogenetic expression and 17ß-estradiol regulation of immune-related genes in early life stages of Japanese medaka (Oryzias latipes).

Accumulating evidence suggests that environmental endocrine disrupting chemicals (EDCs) may exert adverse effects on aquatic organisms via the modulation of immune competence in addition to the endocrine system. However, to date, most studies have been undertaken only on biochemical and histopathological endpoints, and few studies have addressed the role of immune response gene transcript abundance in response to estrogen. In the present study, the ontogenetic expression of immune-related genes, including three complement components (C3-1, C3-2 and Bf/C2), two cytokines (IL-21 and type I IFN [IFN]), lysozyme (LZM), novel immune-type receptor (NITR-18), Ikaros (IK) and ceruloplasmin (CP) were characterized during different developmental periods (from 0 to 28 d post-hatch [dph]) in Japanese medaka. Furthermore, the responses of these genes to natural estrogen (i.e., 17ß-estradiol [E2]) were evaluated. E2 exposure at sublethal concentrations (0.1-10 µg/L) down-regulated the gene expression of C3-1, C3-2, Bf/C2, LZM and CP, while up-regulating the expression of IL-21, IFN, NITR-18 and IK. The results demonstrate a very different trend in gene expression in fish larvae exposed to E2 when compared with the ontogenetic changes in control, suggesting that exposure to environmental chemicals with estrogenic activities may interfere with immune-related genes and thus potentially influence the susceptibility of fish to opportunistic infections. These findings confirm the ability of exogenous estrogens to elicit changes in immune-related gene expression, and broaden our understanding about the mechanisms underlying the actions of EDCs. In addition, the expression profiles of immune-related genes can be developed for use as biomarkers for future immunotoxicological studies.

Environmental factors and genetic background that interact to cause autoimmune thyroid disease.

PURPOSE OF REVIEW: To provide an updated list of genetic and environmental causative factors of autoimmune thyroid disease, and report about the recent discoveries concerning their interaction in the pathogenesis of thyroid autoimmunity. RECENT FINDINGS: Although significant discoveries have been made on genetic and environmental factors underlying the development of autoimmune thyroid disease, few data are available about the mechanisms by which they interact. The most interesting news in this field comes from research on molecular mimicry between microbial antigens and thyroid autoantigens. The molecular mimicry model postulates that, in predisposed subjects, a microbial antigen could trigger autoimmunity because of its structural similarity to an autoantigen of the host, and is a paradigmatic example of the multifactorial interaction of several genes and environmental factors to cause autoimmune diseases, including thyroid diseases. SUMMARY: Recent findings help us to better understand the functional mechanisms of the immune system, which are still only partially known. Beyond the scientific interest, this knowledge has immediate repercussions on clinical practice because it can suggest possible therapeutic targets for new treatments, as well as better and more specific uses of currently available drugs and resources.

Effects of androgen disruption by DDE on the development and functioning of the immune system in Japanese quail.

We hypothesized that immunosuppression in birds that is caused by exposure to antiandrogenic chemicals occurs mainly through disruption of the development of the androgen-sensitive avian lymphoid organ, the bursa of Fabricius. Injections of 20.0 or 40.0 mug of p,p'-DDE [ethylene, 1,1-dichloro-2,2-bis(p-chlorophenyl)], an antiandrogen, were administered at embryonic day 1. Bursas from only chicks treated with DDE were larger than, had fewer follicles, and exhibited vacuolization within follicles compared with controls; spleens were unaffected. No differences in either immune response test were observed. This study demonstrates that the bursa may play a role in androgen-active endocrine disrupting chemical-induced immunosuppression.