Histological and Immunohistochemical Studies to Determine the Mechanism of Cleft Palate Induction after Palatal Fusion in Mice Exposed to TCDD.

Rupture of the basement membrane in fused palate tissue can cause the palate to separate after fusion in mice, leading to the development of cleft palate. Here, we further elucidate the mechanism of palatal separation after palatal fusion in 8-10-week-old ICR female mice. On day 12 of gestation, 40 µg/kg of 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD), sufficient to cause cleft palate in 100% of mice, was dissolved in 0.4 mL of olive oil containing toluene and administered as a single dose via a gastric tube. Fetal palatine frontal sections were observed by H&E staining, and epithelial cell adhesion factors, apoptosis, and cell proliferation were observed from the anterior to posterior palate. TUNEL-positive cells and Ki67-positive cells were observed around the posterior palatal dissection area of the TCDD-treated group. Moreover, in fetal mice exposed to TCDD, some fetuses exhibited cleft palate dehiscence during fusion. The results suggest that palatal dehiscence may be caused by abnormal cell proliferation in epithelial tissues, decreased intercellular adhesion, and inhibition of mesenchymal cell proliferation. By elucidating the mechanism of cleavage after palatal fusion, this research can contribute to establishing methods for the prevention of cleft palate development.

The role of MEG3 in the proliferation of palatal mesenchymal cells is related to the TGFß/Smad pathway in TCDD inducing cleft palate.

Cleft palate (CP) is a common birth defect with a high incidence of occurrence in humans. The 2, 3, 7, 8-tetrachlorodibenzo-p-dioxin (TCDD) is a highly toxic halogenated aromatic hydrocarbon, with a strong CP effect on mice. Increasing recent evidences have shown that long-noncoding RNAs (lncRNAs) play an important role in several diseases, including CP. However, there is a paucity of studies on the role of lncRNA MEG3 in the occurrence and development of TCDD-induced CP. In this study, the relationship between MEG3 and the proliferation of palatal mesenchymal cells and the underlying molecular mechanism were studied by establishing fetal CP with TCDD (64 µg/kg) in C57BL/6N mice. The results revealed that MEG3 was highly expressed during the critical period of CP formation and that the fetal mesenchymal proliferation was significantly inhibited at certain critical periods in the mice receiving TCDD. In addition, we noted a possibility of a crosstalk between MEG3 and the TGF-ß/Smad pathway, such that the inhibition of the TGF-ß/Smad pathway was induced by TCDD. Cumulatively, our study suggests that TCDD-induced CP may be caused by MEG3 inhibition of the proliferation of palatal mesenchymal cells involving the TGFß/Smad pathway, which may provide a novel perspective to understand the pathogenesis of CP.

Quantum mechanical studies on dioxin-imprinted polymer precursor composites: Fundamental insights to enhance the binding strength and selectivity of biomarkers.

We present a benchmark study of binding energies for dioxin-imprinted polymer complexes. A density functional theory approach was used for screening the polymerization precursors in the rational design of molecularly imprinted polymers (MIPs). Tetrachlorodibenzo-p-dioxin (TCDD) was taken as an imprinted molecule. The geometry optimization, natural bond orbital charge, and molecular electrostatic potential of TCDD and acrylamide (AM) were studied at the M062X level and 6-31G(d,p) belonging to one of the hybrid density functional theories. The results of molecular electrostatic potential and natural bond orbital charge analysis were comparable. Among the studied functional monomers-AM, methacrylic acid (MAA), itaconic acid, and vinyl pyridine-AM was confirmed as the best functional monomer, because the strongest interaction (the maximum number of hydrogen bonds and the lowest binding energy) occurs between TCDD and AM. The stability property was excellent when the ratio of TCDD and AM was 1:4. The polarizable continuum model was used for solvent calculations. Computational results showed that acetonitrile plays an important role in the MIP formation, as it seems to control the size and the shape of the cavity. The atoms in molecule and Becke surface method have also been applied to understand the nature and strength of the hydrogen bonding interactions in complexes. TCDD-AM complexes were found involving C-O···Cl and N-H···Cl hydrogen bonds. Good correlations have been established between hydrogen bond lengths versus atoms in molecule topological parameter like electron density ρ(r) and its Laplacian ▽2 ρ(r) at the bond critical points. On ground of theoretical results, a series of MIPs were synthesized. The MIP prepared using TCDD as the template, the functional monomer (AM), and the cross-linker (TRIM) in acetonitrile solvent exhibited the highest adsorption capacity for TCDD. The maximum binding capacity of TCDD on the MIP was 3.7 µg/mg. This research work can provide a theoretical reference for the fabrication and characterization of novel TCDD-MIPs for environmental applications.

Chemical indicators of exposure to polychlorodibenzo-p-dioxins, polychlorodibenzofurans and polychlorobiphenyls in breast milk samples from mothers residing in Trento, Italy, and neighboring country municipalities.

A human biomonitoring study was carried out in the province of Trento, northern Italy, on two groups of women residing in areas with presumably different levels of exposure to polychlorodibenzo-p-dioxins (PCDDs), polychlorodibenzofurans (PCDFs) and polychlorobiphenyls (PCBs). The aim was characterization of the possible impact of a steel plant and an urban environment on the internal doses of these contaminants. The areas investigated were the following: (i) the municipality of Borgo Valsugana, where a steel plant has been operating for the last 30 years, and (ii) the city of Trento. Ten women were enrolled from each area, and breast milk samples were collected from each woman. The amounts of PCDDs, PCDFs and PCBs were analyzed in all samples, and the data obtained were evaluated by descriptive and multivariate statistical approaches. For all samples, the concentrations observed were in the current acceptable concentration range observed in Europe. The descriptive statistics, however, clearly showed that some differences existed between the two groups: the PCDD, PCDF and DL-PCB concentrations were consistently higher in the Trento group of women living in the area surrounding the steel plant. Congener-profile analysis was carried out on both groups, and an appropriate multivariate approach, such as classical factor analysis (CFA), was used to investigate possible differential exposure sources. The application of this approach made it possible to show the quantitative and qualitative differences that characterized the two areas.

Autopsy tissues as biological monitors of human exposure to environmental pollutants. A case study: Concentrations of metals and PCDD/Fs in subjects living near a hazardous waste incinerator.

Human biomonitoring is of tremendous importance to prevent potential adverse effects derived from human exposure to chemicals. Blood and urine are among the biological monitors more frequently used. However, biological matrices such as breast milk, hair, nails, saliva, feces, teeth, and expired air are also often used. In addition, and focused mainly on long-term exposure, adipose tissue and other human tissues like bone, liver, brain or kidney, are also used as biological monitors of certain substances, especially for long-term biomonitoring. However, for this kind of tissues sampling is always a limiting factor. In this paper, we have examined the role of autopsy tissues as biological monitors of human exposure to environmental pollutants. For it, we have used a case study conducted near a hazardous waste incinerator (HWI) in Catalonia (Spain), in which the concentrations of metals and polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs), have been periodically determined in autopsy tissues of subjects living in the area under potential influence of the facility. This case study does not show advantages -in comparison to other appropriate biomonitors such as blood- in using autopsy tissues in the monitoring of long-term exposure to metals and PCDD/Fs.

LncRNA H19 and Target Gene-mediated Cleft Palate Induced by TCDD.

This study investigated the role of long non-coding RNAs (lncRNAs) in the development of the palatal tissues. Cleft palates in mice were induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Expression levels of long non-coding RNA H19 (lncRNA H19) and insulin-like growth factor 2 (IGF2) gene were measured by quantitative real-time polymerase chain reaction (qRT-PCR). The rate of occurrence of cleft palate was found to be 100% by TCDD exposure, and TCDD could cause short upper limb, cerebral fissure, webbed neck, and short neck. The expression levels of lncRNA H19 and IGF2 gene specifically showed embryo age-related differences on E13, E14, and E15 in the palatal tissues. The expression levels of lncRNA H19 and IGF2 gene showed an inverse relationship on E13, E14, and E15. These findings demonstrated that lncRNA H19 and IGF2 can mediate the development of mouse cleft palate.

Tooth development disorders in infants of rat dams exposed to 2,3,7,8-tetrachlorodibenzo-p-dioxin and protective role of tocopherol and acetylsalicylic acid.

Aryl hybrocardon receptor (AhR) activation plays a key role in the pathomechanism of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)-induced defective spatial structure of teeth caused by disordered collagen synthesis. The aim of this study was to identify the influence of dioxins present in female Buffalo rats on the dental structure of their offspring's in the neonatal period and the potential of α-tocopherol and acetylsalicylic acid in curbing post-dioxin hard tissue defects. Research material consisted of molar teeth (n=40) of rat pups which had been given a single dose of TCDD and were then treated with tocopherol or acetylsalicylic acid for 3 weeks. In the offspring of rat dams exposed to TCDD, ameloblasts and odontoblasts were less developed in comparison with the control group and less dynamic angiogenesis in the area of dental papilla was observed. In the pups of TCDD-exposed mothers, a smaller number of AhR was found in amelogenic and odontoblastic cells, whereas in the pups of mothers exposed to TCDD followed by tocopherol and acetylsalicylic acid treatment, the expression of AhR in ameloblasts and odontoblasts increased. We conclude that tocopherol and acetylsalicylic acid treatment exerts a protective effect on the TCDD-induced structural defects of tooth tissue.

Effect of TCDD on the fate of epithelial cells isolated from human fetal palatal shelves (hFPECs).

Cleft palate is caused by the failure of palatal midline epithelial cells to disintegrate, which is necessary for palatal mesenchymal confluence. Although 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) exposure is linked to cleft palate at a high rate, the mechanism remains to be elucidated. The present study was designed to determine the effects of TCDD on the fate of epithelial cell isolated from human fetal palatal shelves (hFPECs). We demonstrate that TCDD increased cell proliferation and promoted the progression of cells from G1 to S phase as well as increased the number of cells entering the G2/M phase. We found that TCDD has no measurable effect on apoptosis of hFPECs. The protein level assays revealed that TCDD increased cyclin-dependent kinases 4 (cdk4), cyclin D1, cyclin E and p21 (Waf1/Cip1) but not cdk2, bcl-2, cyclin B1 and cyclin A. Furthermore, TCDD activated PI3K/AKT signaling, and the PI3K inhibitor, LY294002, partially abrogated TCDD-induced cell proliferation and gene modulations. TCDD treatment increased CYP1A1 mRNA and protein levels, which indicated the activation of AhR signaling. Knockdown of the AhR with siRNA suppressed TCDD-induced cell proliferation and PI3K/AKT signaling activation. Taken together, these data demonstrated that TCDD is able to promote growth of hFPECs through AhR-dependent activation of the PI3K/AKT pathway, which may account for the underlying mechanism by which TCDD causes a failure of palatal fusion.

TCDD promoted EMT of hFPECs via AhR, which involved the activation of EGFR/ERK signaling.

One critical step of second palatal fusion is the newly formed medial epithelia seam (MES) disintegration, which involves apoptosis, epithelial to mesenchymal transition (EMT), and cell migration. Although the environmental toxicant 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) produces cleft palate at high rates, little is known about the effects of TCDD exposure on the fate of palatal epithelial cells. By using primary epithelial cells isolated from human fetal palatal shelves (hFPECs), we show that TCDD increased cell proliferation and EMT, as demonstrated by increased the epithelial markers (E-cadherin and cytokeratin14) and enhanced the mesenchymal markers (vimentin and fibronectin), but had no effect on cell migration and apoptosis. TCDD exposure led to a dose-dependent increase in Slug protein expression. Coimmunoprecipitation revealed that TCDD promoted AhR to form a protein complex with Slug. ChIP assay confirmed that TCDD exposure recruited AhR to the xenobiotic responsive element of Slug promoter. Knockdown of AhR by siRNA remarkably weakened TCDD-induced binding of AhR to the XRE promoter of slug, thereby suppressed TCDD-induced vimentin. Further experiment showed that TCDD stimulated EGFR phosphorylation did not influence the TGFß3/Smad signaling; whereas TCDD increased phosphorylation of ERK1/2 and p38 with no effect on activation of JNK. By using varieties of inhibitors, we confirmed that TCDD promoted proliferation and EMT of hFPECs via activation of EGFR/ERK pathway. These data make a novel contribution to the molecular mechanism of cleft palate by TCDD.

Experimental Solubility Approach to Determine PDMS-Water Partition Constants and PDMS Activity Coefficients.

Freely dissolved aqueous concentration and chemical activity are important determinants of contaminant transport, fate, and toxic potential. Both parameters are commonly quantified using Solid Phase Micro-Extraction (SPME) based on a sorptive polymer such as polydimethylsiloxane (PDMS). This method requires the PDMS-water partition constants, KPDMSw, or activity coefficient to be known. For superhydrophobic contaminants (log KOW >6), application of existing methods to measure these parameters is challenging, and independent measures to validate KPDMSw values would be beneficial. We developed a simple, rapid method to directly measure PDMS solubilities of solid contaminants, SPDMS(S), which together with literature thermodynamic properties was then used to estimate KPDMSw and activity coefficients in PDMS. PDMS solubility for the test compounds (log KOW 7.2-8.3) ranged over 3 orders of magnitude (4.1-5700 µM), and was dependent on compound class. For polychlorinated biphenyls (PCBs) and polychlorinated dibenzo-p-dioxins (PCDDs), solubility-derived KPDMSw increased linearly with hydrophobicity, consistent with trends previously reported for less chlorinated congeners. In contrast, subcooled liquid PDMS solubilities, SPDMS(L), were approximately constant within a compound class. SPDMS(S) and KPDMSw can therefore be predicted for a compound class with reasonable robustness based solely on the class-specific SPDMS(L) and a particular congener's entropy of fusion, melting point, and aqueous solubility.

Effect of colestimide on the concentrations of polychlorinated dibenzo-p-dioxins, polychlorinated dizenzofurans, and polychlorinated biphenyls in blood of Yusho patients.

BACKGROUND: Oral colestimide was reported to lower the concentration of PCDDs, PCDFs, and PCB in the blood of humans. A pilot study showed that the arithmetic mean total TEQ concentrations of PCDDs, PCDFs, and PCBs in the blood of subjects after the trial decreased approximately 20 % compared to pre-trial levels, suggesting that colestimide could decrease human dioxin levels. We designed the current clinical trial study based on this information. In this study, we examined whether colestimide could reduce the individual congener concentrations of PCDDs, PCDFs, and PCBs in the blood of Yusho patients. METHODS: Out of the 36 Yusho patients who participated in the clinical trial, 26 patients self-administered colestimide 3 g/day orally for 6 months. The concentrations of PCDDs, PCDFs and PCBs in the blood of 26 Yusho patients before the trial were compared with those after the trial. RESULTS: The arithmetic mean total TEQ concentrations of PCDDs, PCDFs, non-ortho PCBs, and mono-ortho PCBs in the blood of the 26 Yusho patients before and after the clinical trial were 42-303 (mean: 130, median: 120) and 43-283 (mean: 132, median: 118) pg TEQ/g lipid, respectively. The sums of the concentrations of 58 PCB congeners measured in the blood of Yusho patients before and after the trial were 321-2643 (mean: 957, median: 872) and 286-2007 (mean: 975, median: 806) ng/g lipid, respectively, indicating that the concentrations of PCDDs, PCDFs, and PCBs after the trial were almost the same as those before the trial. Among congeners of PCDDs, PCDFs, dioxin-like PCBs, and non-dioxin-like PCBs, most congeners of these compounds did not show a statistically significant decrease after the trial. CONCLUSION: Colestimide may not be beneficial in reducing the high blood levels of dioxin-like compounds in Yusho patients.

Assessing polychlorinated dibenzo-p-dioxins and polychlorinated dibenzofurans in air across Latin American countries using polyurethane foam disk passive air samplers.

A passive air sampling network has been established to investigate polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs) at Global Atmospheric Passive Sampling (GAPS) sites and six additional sites in the Group of Latin American and Caribbean Countries (GRULAC) region. The air sampling network covers background, agricultural, rural, and urban sites. Samples have been collected over four consecutive periods of 6 months, which started in January 2011 [period 1 (January to June 2011), period 2 (July to December 2011), period 3 (January to June 2012), and period 4 (July 2012 to January 2013)]. Results show that (i) the GAPS passive samplers (PUF disk type) and analytical methodology are adequate for measuring PCDD/F burdens in air and (ii) PCDD/F concentrations in air across the GRULAC region are widely variable by almost 2 orders of magnitude. The highest concentrations in air of Σ4-8PCDD/Fs were found at the urban site São Luis (Brazil, UR) (i.e., 2560 fg/m3) followed by the sites in São Paulo (Brazil, UR), Mendoza (Argentina, RU), and Sonora (Mexico, AG) with values of 1690, 1660, and 1610 fg/m3, respectively. Very low concentrations of PCDD/Fs in air were observed at the background site Tapanti (Costa Rica, BA), 10.8 fg/m3. This variability is attributed to differences in site characteristics and potential local/regional sources as well as meteorological influences. The measurements of PCDD/Fs in air agree well with model-predicted concentrations performed using the Global EMEP Multimedia Modeling System (GLEMOS) and emission scenario constructed on the basis of the UNEP Stockholm Convention inventory of dioxin and furan emissions.

Two Decades of Environmental Surveillance in the Vicinity of a Waste Incinerator: Human Health Risks Associated with Metals and PCDD/Fs.

The concentrations of polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs), as well as the levels of a number of heavy metals, have been periodically measured in samples of soil and vegetation collected around a municipal solid waste incinerator (MSWI) in Tarragona (Catalonia, Spain) for approximately 20 years. Since 2007, the levels of the above-mentioned pollutants have also been determined in air samples by means of either active or passive samplers. In the present study, data regarding the environmental impact of the MSWI, in terms of PCDD/Fs and heavy metals, are updated. The temporal trends of these pollutants were evaluated by comparison with data from previous surveys. In the current survey (2013-2014), mean concentrations of PCDD/Fs in soil, vegetation, and air were 0.63 ng I-TEQ/g, 0.07 ng I-TEQ/g, and 10.1 fg WHO-TEQ/m(3), respectively. Decreases of 47 and 35 % of PCDD/Fs in soil and vegetation, respectively, were observed in relation to the background study (1999). Regarding air samples, a slight temporal decrease of the PCDD/F levels was also found with the remaining concentrations staying nearly constant through time. With respect to metals, notable fluctuations in the concentrations were noted, being dependent on each specific environmental monitor. Overall, the current exposure to PCDD/Fs and metals does not mean any additional health risks for the population living near the facility. In conclusion, the results of the present study show that the environmental impact of the Tarragona MSWI is not significant, in terms of PCDD/Fs and heavy metals, after >20 years of continuous operation.

PCDD/Fs in the soils in the province of Trento: 10 years of monitoring.

This paper presents a 10-year overview of the dioxins and furans (PCDD/Fs) content in soils in the province of Trento (Italy). The aim was to compare the results found in the Valsugana valley where there is a steel-making plant with other locations within the province. During 2002 and from 2005 to 2010, campaigns were carried out in order to obtain a background reference in terms of micropollutants, in view of the possible construction of a municipal solid waste (MSW) incinerator in Trento. In 2009, a campaign was performed for the environmental characterization of the Valsugana valley, the town of Trento and its surroundings, in order to help assess the impact of the steel-making plant. In 2012, another campaign was carried out by the Department of Civil and Environmental Engineering of Trento, in order to monitor the soils in the area around the steel mill. All the campaigns showed relatively low concentrations of PCDD/Fs, both in protected areas and in the areas close to the industrial plants. No critical situations were identified, as also confirmed by an estimation of the potential daily PCDD/F intake by children subject to accidental ingestion of soil.

A new approach for assessing the state of environment using isometric log-ratio transformation and outlier detection for computation of mean PCDD/F patterns in biota.

To assess the state of the environment, various compartments are examined as part of monitoring programs. Within monitoring, a special focus is on chemical pollution. One of the most toxic substances ever synthesized is the well-known dioxin 2,3,7,8-TCDD (2,3,7,8-tetra-chlor-dibenzo-dioxin). Other PCDD/F (polychlorinated-dibenzo-dioxin and furan) can act toxic too. They are ubiquitary and persistent in various environmental compartments. Assessing the state of environment requires knowledge of typical local patterns of PCDD/F for as many compartments as possible. For various species of wild animals and plants (so called biota), I present the mean local congenere profiles of ubiquitary PCDD/F contamination reflecting typical patterns and levels of environmental burden for various years. Trends in time series of means can indicate success or failure of a measure of PCDD/F reduction. For short time series of mean patterns, it can be hard to detect trends. A new approach regarding proportions of outliers in the corresponding annual cross-sectional data sets in parallel can help detect decreasing or increasing environmental burden and support analysis of time series. Further, in this article, the true structure of PCDD/F data in biota is revealed, that is, the compositional data structure. It prevents direct application of statistical standard procedures to the data rendering results of statistical analysis meaningless. Results indicate that the compositional data structure of PCDD/F in biota is of great interest and should be taken into account in future studies. Isometric log-ratio (ilr) transformation is used, providing data statistical standard procedures that can be applied too. Focusing on the identification of typical PCDD/F patterns in biota, outliers are removed from annual data since they represent an extraordinary situation in the environment. Identification of outliers yields two advantages. First, typical (mean) profiles and levels of PCDD/F contamination can be identified. Second, decreasing (increasing) proportions of outliers could indicate decreasing (increasing) numbers of extraordinary environmental burden rendering the success of PCDD/F reduction strategies for biota. Therefore, probabilities and proportions of outlier contamination are estimated too. To reveal the enormous influence of the method of outlier detection, the applied two well-known procedures are compared, that is, robust Mahalanobis distance and a projection pursuit-based approach.

Agent Orange exposure and cancer incidence in Korean Vietnam veterans: a prospective cohort study.

BACKGROUND: During the Vietnam War, US and allied military sprayed approximately 77 million liters of tactical herbicides including Agent Orange, contaminated with 2,3,7,8-tetrachlorodibenzo-p-dioxin. To the authors' knowledge, few studies to date have examined the association between Agent Orange exposure and cancer incidence among Korean veterans who were exposed to Agent Orange during the Vietnam War. METHODS: An Agent Orange exposure index, based on the proximity of the veteran's military unit to the area that was sprayed with Agent Orange, was developed using a geographic information system-based model. Cancer incidence was followed for 180,251 Vietnam veterans from 1992 through 2003. RESULTS: After adjustment for age and military rank, high exposure to Agent Orange was found to significantly increase the risk of all cancers combined (adjusted hazards ratio [aHR], 1.08). Risks for cancers of the mouth (aHR, 2.54), salivary glands (aHR, 6.96), stomach (aHR, 1.14), and small intestine (aHR, 2.30) were found to be significantly higher in the high-exposure group compared with the low-exposure group. Risks for cancers of all sites combined (aHR, 1.02) and for cancers of the salivary glands (aHR, 1.47), stomach (aHR, 1.03), small intestine (aHR, 1.24), and liver (aHR, 1.02) were elevated with a 1-unit increase in the exposure index. CONCLUSIONS: Exposure to Agent Orange several decades earlier may increase the risk of cancers in all sites combined, as well as several specific cancers, among Korean veterans of the Vietnam War, including some cancers that were not found to be clearly associated with exposure to Agent Orange in previous cohort studies primarily based on Western populations.

Aryl hydrocarbon receptor-mediated induction of EBV reactivation as a risk factor for Sjögren's syndrome.

The aryl hydrocarbon receptor (AhR) is a ligand-activated transcription factor that mediates a variety of biological effects by binding to environmental pollutants, including 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD or dioxin). Although numerous animal studies have demonstrated the harmful effects of dioxins, it remains controversial whether dioxins pose a risk to human health. Enhanced lytic replication of EBV is a risk factor for the development of autoimmune diseases and cancers. This study evaluated the possibility that ligand-activated AhR reactivates EBV. EBV reactivation and AhR transactivation were evaluated with luciferase assays. Saliva samples were collected from 19 patients with primary Sjögren's syndrome (SS). Control saliva samples were obtained from 10 healthy individuals and nine patients with severe dry mouth. TCDD enhanced BZLF1 transcription, which mediates the switch from the latent to the lytic form of EBV infection in EBV-positive B cell lines and in a salivary gland epithelial cell line. Moreover, TCDD-induced increases in BZLF1 mRNA and EBV genomic DNA levels were confirmed in the B cell lines. Saliva from SS patients activated the transcription of both CYP1A1 and BZLF1. Additionally, there was a positive correlation between CYP1A1 and BZLF1 promoter activities. AhR ligands elicited the reactivation of EBV in activated B cells and salivary epithelial cells, and these ligands are involved in SS. Our findings reveal novel aspects of the biological effects of dioxin and the AhR-dependent pathogenesis of autoimmune diseases.

Deposition fluxes of PCDD/Fs in the area surrounding a steel plant in northwest Italy.

The paper aims at investigating the contribution of a steel plant located in a rural area in northwestern Italy (700,000 tons of steel/year) to the deposition fluxes of Polychorinated Dibenzo-p-dioxins and Polychorinated dibenzofurans (PCDD/PCDFs) at local level through the analysis of sampling data, literature data, and air dispersion model (AERMOD)output data. Total measured deposition fluxes of PCDD/PCDFs in three monitoring stations were consistent with other studies carried out in Italy in urban and suburban areas and in rural European areas; while these were lower than those measured in other European urban/suburban areas or in sites influenced by industrial sources. Furthermore, the measured fluxes were also compared with the pattern of PCDD/Fs in ambient air sampled at the same sites in a previous study. This comparison showed a similarity between air concentration and deposition patterns of the samples collected at the three monitoring stations and a clear distinction of these from the source. The study was completed with AERMOD simulations, conducted with a mass mean particle diameter of 0.5 µm, according to the particle size distribution of the samples collected at the source. AERMOD calculated deposition fluxes of two to three orders of magnitude lower than those measured in two monitoring points; while in the most distant monitoring station, the deposition fluxes were too low to be calculated by the model. The simulations confirmed that the most distant monitoring station was not subject to emissions from the steel plant. The analysis highlighted the limited influence of the source in the local PCDD/F deposition fluxes.

Selective pressurized liquid extraction technique capable of analyzing dioxins, furans, and PCBs in clams and crab tissue.

A selective pressurized liquid extraction technique (SPLE) was developed for the analysis of polychlorodibenzo-p-dioxins, polychlorodibenzofurans (PCDD/Fs) and dioxin-like polychlorobiphenyls (dl-PCBs) in clam and crab tissue. The SPLE incorporated multiple cleanup adsorbents (alumina, florisil, silica gel, celite, and carbopack) within the extraction cell. Tissue extracts were analyzed by high resolution gas chromatography coupled with electron capture negative ionization mass spectrometry. Mean recovery (n = 3) and percent relative standard deviation for PCDD/Fs and dl-PCBs in clam and crabs was 89 ± 2.3 and 85 ± 4.0, respectively. The SPLE method was applied to clams and crabs collected from the San Jacinto River Waste Pits, a Superfund site in Houston, TX. The dl-PCBs concentrations in clams and crabs ranged from 50 to 2,450 and 5 to 800 ng/g ww, respectively. Sample preparation time and solvents were reduced by 92 % and 65 %, respectively, as compared to USEPA method 1613.

Combined toxic effects of polystyrene microplastics and 3,6-dibromocarbazole on zebrafish (Danio rerio) embryos.

Microplastics (MPs) and polyhalogenated carbazoles (PHCZs) are widely detected in the aquatic environment, and their ecological risks have become a research focus. Although there is an extensive co-distribution of MPs and PHCZs, their combined toxicity to aquatic organisms is still unclear. This study investigated the toxic effects of polystyrene microplastics (PS-MPs) and 3,6-dibromocarbazole (3,6-DBCZ) on zebrafish embryos by individual/combined exposure. This study showed that individual or combined exposure of PS-MPs (10 mg/L) and 3,6-DBCZ (0.5 mg/L) could significantly increase the rate of zebrafish embryo deformity, whereas no significant effect was observed on mortality and hatching rate. Furthermore, exposure to 3,6-DBCZ or PS-MPs increased reactive oxygen species (ROS) levels in zebrafish embryos, and the resulting oxidative stress induced apoptosis. Comparably, the levels of oxidative stress and apoptosis in zebrafish embryos were significantly reduced with the combined exposure of 3,6-DBCZ and PS-MPs. These observations suggest that the combined exposure of 3,6-DBCZ and PS-MPs has an antagonistic effect on oxidative stress and apoptosis. Fluorescence PS-MPs tracing and 3,6-DBCZ enrichment analysis showed that, with the protection of chorion, the entry of PS-MPs (5 and 50 µm) into the embryonic stage (55 hpf) of zebrafish was prevented. Moreover, after exposure for 96-144 hpf, PS-MPs served as a carrier to promote the 3,6-DBCZ accumulation and its dioxin-like toxicity in zebrafish larvae through ingestion. Compared with 5-µm PS-MPs, 50-µm PS-MPs promoted higher accumulation and dioxin-like toxicity of 3,6-DBCZ in zebrafish larvae. These findings provide that MPs can be used as an important carrier of PHCZs, influencing their toxicity and bioaccumulation in the organisms.