Analysis of predictors of fever after aortic valve replacement: Diabetic patients are less likely to develop fever after aortic valve replacement, a single-centre retrospective study.

BACKGROUND: Postoperative temperature dysregulation affects the length of hospital stay and prognosis. This study evaluated the factors that influence the occurrence of fever in patients after aortic valve replacement surgery. METHODS: Eighty-seven consecutive patients who underwent aortic valve replacement surgery were included. Patients' age, sex and body mass index; presence of diabetes mellitus; operation time; blood loss; blood transfusion volume; preoperative and postoperative laboratory findings; presence or absence of oral function management; and fever >38°C were retrospectively analysed through univariate and multiple logistic regression analyses. RESULTS: Among the variables, only diabetes mellitus status was significantly associated with fever ⩾38°C. Postoperatively, patients with diabetes mellitus were significantly less likely to develop fever above 38°C and a fever rising to 38°C. CONCLUSIONS: This study shows that the presence of comorbid diabetes mellitus decreases the frequency of developing fever >38°C after aortic valve replacement surgery.

Transcriptome analysis reveals limited toxic effects of the UV-filter benzophenone-3 (BP-3) on the hermatypic coral Acropora tenuis and its symbiotic dinoflagellates.

This study aimed to investigate the toxic and transcriptomic effects of the ultraviolet filter benzophenone-3 (BP-3) on Acropora tenuis and its symbiotic dinoflagellates while using acetone as a solvent. Seven-day exposure to 50 and 500 µg/L, which is higher than most BP-3 records from coastal waters, did not affect coral colour or dinoflagellate photosynthesis. Differentially expressed genes (DEGs) between seawater and solvent controls were <20 in both corals and dinoflagellates. Eleven coral DEGs were detected after treatment with 50 µg/L BP-3. Fourteen coral DEGs, including several fluorescent protein genes, were detected after treatment with 500 µg/L BP-3. In contrast, no dinoflagellate DEGs were detected in the BP-3 treatment group. These results suggest that the effects of 50-500 µg/L BP-3 on adult A. tenuis and its dinoflagellates are limited. Our experimental methods with lower acetone toxicity provide a basis for establishing standard ecotoxicity tests for corals.

Elucidating Key Characteristics of PFAS Binding to Human Peroxisome Proliferator-Activated Receptor Alpha: An Explainable Machine Learning Approach.

Per- and polyfluoroalkyl substances (PFAS) are widely employed anthropogenic fluorinated chemicals known to disrupt hepatic lipid metabolism by binding to human peroxisome proliferator-activated receptor alpha (PPARα). Therefore, screening for PFAS that bind to PPARα is of critical importance. Machine learning approaches are promising techniques for rapid screening of PFAS. However, traditional machine learning approaches lack interpretability, posing challenges in investigating the relationship between molecular descriptors and PPARα binding. In this study, we aimed to develop a novel, explainable machine learning approach to rapidly screen for PFAS that bind to PPARα. We calculated the PPARα-PFAS binding score and 206 molecular descriptors for PFAS. Through systematic and objective selection of important molecular descriptors, we developed a machine learning model with good predictive performance using only three descriptors. The molecular size (b_single) and electrostatic properties (BCUT_PEOE_3 and PEOE_VSA_PPOS) are important for PPARα-PFAS binding. Alternative PFAS are considered safer than their legacy predecessors. However, we found that alternative PFAS with many carbon atoms and ether groups exhibited a higher affinity for PPARα. Therefore, confirming the toxicity of these alternative PFAS compounds with such characteristics through biological experiments is important.

Expression analyses of stress-responsive genes in the hermatypic coral Acropora tenuis and its symbiotic dinoflagellates after exposure to the herbicide Diuron.

Diuron is one of the most frequently applied herbicides in sugarcane farming in southern Japan, and Australia. In addition, it is used as a booster substance in copper-based antifouling paints. Due to these various uses, Diuron is released into the marine environment; however, little information is available on gene expression in corals and their symbiotic algae exposed to Diuron. We investigated the effects of Diuron on stress-responsive gene expression in the hermatypic coral Acropora tenuis and its symbiotic dinoflagellates. After seven days of exposure to 1 µg/L and 10 µg/L Diuron, no significant changes in the body colour of corals were observed. However, quantitative reverse transcription-polymerase chain reaction analyses revealed that the expression levels of stress-responsive genes, such as heat shock protein 90 (HSP90), HSP70, and calreticulin (CALR), were significantly downregulated in corals exposed to 10 µg/L of Diuron for seven days. Moreover, aquaglyceroporin was significantly downregulated in corals exposed to environmentally relevant concentrations of 1 µg/L Diuron. In contrast, no such effects were observed on the expression levels of other stress-responsive genes, such as oxidative stress-responsive proteins, methionine adenosyltransferase, and green/red fluorescent proteins. Diuron exposure had no significant effect on the expression levels of HSP90, HSP70, or HSP40 in the symbiotic dinoflagellates. These results suggest that stress-responsive genes, such as HSPs, respond differently to Diuron in corals and their symbiotic dinoflagellates and that A. tenuis HSPs and CALRs may be useful molecular biomarkers for predicting stress responses induced by the herbicide Diuron. Supplementary Information: The online version contains supplementary material available at 10.1007/s42995-023-00183-0.

Adverse effects of contamination by fipronil and its derivatives on growth, molting, and gene expression in the mysid crustacean, Americamysis bahia, in Japanese estuaries.

Concentrations of fipronil (Fip) and several of its derivatives were detected in samples from four rivers and four estuaries in Japan. LC-MS/MS analysis detected Fip and its derivatives, except for fipronil detrifluoromethylsulfinyl, in almost all samples. The total concentrations of the five compounds were approximately two-fold greater in river water (mean: 21.2, 14.1, and 9.95 ng/L in June, July, and September, respectively) compared to those in estuarine water (mean: 10.3, 8.67, and 6.71 ng/L, respectively). Fipronil, fipronil sulfone (Fip-S), and fipronil sulfide (Fip-Sf) represented more than 70 % of all compounds. This is the first report to demonstrate the contamination of estuarine waters of Japan by these compounds. We further investigated the potentially toxic effects of Fip, Fip-S, and Fip-Sf on the exotic mysid, Americamysis bahia (Crustacea: Mysidae). The lowest effective concentrations of Fip-S (10.9 ng/L) and Fip-Sf (19.2 ng/L) on mysid growth and molting was approximately 12.9- and 7.3-fold lower than Fip (140.3 ng/L), suggesting they had higher toxicity. Quantitative reverse transcription polymerase chain reaction analysis revealed that ecdysone receptor and ultraspiracle gene expression were not affected after 96-h of exposure to Fip, Fip-S, and Fip-Sf, suggesting that these genes may not be involved in the molting disruption induced by Fip, Fip-S, and Fip-Sf. Our findings suggest that environmentally relevant concentrations of Fip and its derivatives can disrupt the growth of A. bahia by promoting molting. However, further studies are required to elucidate its molecular mechanism.

Developmental toxicity and transcriptome analysis of equine estrogens in developing medaka (Oryzias latipes) using nanosecond pulsed electric field incorporation.

Equine estrogens (EQs) are steroidal hormones isolated from the urine of pregnant mares and are used in the formulation of human medications. This study initially investigated the embryonic developmental toxicity of equilin (Eq) and equilenin (Eqn) in medaka (Oryzias latipes). Malformations were observed in embryos exposed to nominal concentrations of 1 and 10 mg/L of Eq and Eqn. Delayed hatching was observed at 1 mg/L of Eq. To further investigate the molecular mechanism of developmental toxicity caused by Eq and Eqn, transcriptome and bioinformatics analyses were performed. Among 2016 and 3855 total differentially expressed genes (DEGs), 1117 DEGs overlapped between Eq. (55.4 % of total DEGs) and Eq. (29.0 % of total DEGs). Gene ontology indicated effects in terms related to blood circulation and cell junctions. Pathway analyses using DEGs revealed that both Eq and Eqn treatments at 10 mg/L affected various KEGG (Kyoto Encyclopedia of Genes and Genomes) pathways, such as neuroactive ligand-receptor interaction, mitogen-activated protein kinase signaling, retinol metabolism, and cytokine-cytokine receptor interaction. These results suggest that the disruption of these KEGG pathways is involved in the developmental toxicity of EQs in medaka embryos.

Efficient in vivo and in silico assessments of antiandrogenic potential in zebrafish.

This study aimed to establish zebrafish-based in vivo and in silico assay systems to evaluate the antiandrogenic potential of environmental chemicals. Zebrafish embryos were exposed to 17α-methyltestosterone (TES) alone or coexposed to TES and representative antiandrogens including flutamide, p,p'-DDE, vinclozolin, fenitrothion, and linuron. We assessed the transcript expression of the androgen-responsive gene sulfotransferase family 2, cytosolic sulfotransferase 3 (sult2st3). The expression of sult2st3 was significantly induced by TES in the later stages of embryonic development. However, the TES-induced expression of sult2st3 was inhibited by flutamide in a concentration-dependent manner (IC50: 5.7 µM), suggesting that the androgen receptor (AR) plays a role in sult2st3 induction. Similarly, p,p'-DDE, vinclozolin, and linuron repressed the TES-induced expression of sult2st3 (IC50s: 0.35, 3.9, and 52 µM, respectively). At the highest concentration tested (100 µM), fenitrothion also suppressed sult2st3 expression almost completely. Notably, p,p'-DDE and linuron did not inhibit sult2st3 induction due to higher concentrations of TES; instead, they potentiated TES-induced sult2st3 expression. Fenitrothion and linuron, which had relatively low antiandrogenic potentials in terms of sult2st3 inhibition, induced broader toxicities in zebrafish embryos; thus, the relationship between developmental toxicities and antiandrogenic potency was unclear. Additionally, an in silico docking simulation showed that all five chemicals interact with the zebrafish AR at relatively low interaction energies and with Arg702 as a key amino acid in ligand binding. Our findings suggest that a combination of zebrafish-based in vivo and in silico assessments represents a promising tool to assess the antiandrogenic potentials of environmental chemicals.

Assessment of binding potencies of polychlorinated biphenyls and polybrominated diphenyl ethers with Baikal seal and mouse constitutive androstane receptors: Comparisons across species and congeners.

The present study evaluated the binding potencies (equilibrium dissociation constant: KD) of polychlorinated biphenyls (PCBs) and polybrominated diphenyl ethers (PBDEs) with the constitutive androstane receptor (CAR)_ligand binding domain (LBD) of the Baikal seal (bsCAR_LBD) and mouse (mCAR_LBD) using a surface plasmon resonance (SPR) biosensor. The binding affinities of individual congeners with mCAR_LBD tended to be higher than those with bsCAR_LBD but the differences were within the same order of magnitude. Notably, PBDE congeners showed higher binding affinities for both CAR_LBDs than PCB congeners. In silico docking simulations demonstrated that PBDEs had more non-covalent interactions with specific amino acid residues in both CAR_LBDs than PCBs, supporting the results of their binding affinities. Binding affinity comparisons among congeners revealed the structural requirements for higher binding; mono or di ortho-, tri meta-, and di para­chlorine substitutions for PCBs, and di or tri ortho-, mono meta-, and di para­bromine substitutions for PBDEs. The binding potencies of these congeners unlikely accounted for their previously reported CAR-mediated transactivation potencies, implying that their transactivation is regulated in a ligand-dependent, but a distinct manner from ligand binding. Risk assessment analysis showed that the KD values of individual PCB and PBDE congeners were 1-4 orders of magnitude higher than their respective hepatic concentrations in wild Baikal seal population.

Effects of the herbicide Irgarol 1051 on the transcriptome of hermatypic coral Acropora tenuis and its symbiotic dinoflagellates.

Coral reefs face multiple threats, including climate change, agricultural runoff, shipping activities, coastal development, and chemical pollutants. Irgarol 1051, a PSII herbicide, has been used as an antifouling booster since the previously used antibiofouling agent tributyltin (TBT) was banned worldwide. Although the mechanisms through which elevated temperatures cause coral bleaching have been reported, it remains unclear how PSII herbicides cause bleaching. Thus, in this study, we investigated the transcriptomes of Acropora tenuis and its symbiotic dinoflagellates by RNA-sequencing (RNA-Seq) to elucidate the molecular mechanisms underlying Irgarol-induced bleaching. Coral exposure to 10 µg/L Irgarol for 7 d affected coral body colour, specifically by an increase in their red, green, and blue (RGB) values; however, no such effect was observed in corals exposed to 1 µg/L Irgarol. RNA-Seq revealed the differentially expressed genes (DEGs) in corals and symbiotic dinoflagellates following Irgarol exposure. Coral DEGs encoded green fluorescent protein, blue-light-sensing photoreceptor (cryptochrome), chromoprotein, caspase 8, and nuclear receptors; DEGs in symbiotic dinoflagellates encoded light-harvesting proteins, photosystem II proteins, and heat shock proteins (i.e. HSP70 and HSP90), and ubiquitin. Bioinformatic analyses revealed that both Irgarol treatments disrupted various gene ontology terms, pathways, and protein interaction networks; these are different in corals (e.g. oxidative phosphorylation, metabolic pathway, transforming growth factor-ß signalling pathway, adherens junction, and apoptosis) and symbiotic dinoflagellates (e.g. protein processing in endoplasmic reticulum, carbon fixation in photosynthetic organisms, metabolic pathway, and photosynthesis). Our data suggest that Irgarol disrupts the expression of various coral genes, thereby affecting various gene ontology terms, pathways, and protein interaction networks. Our study provides new insights into the potential molecular mechanisms underlying the bleaching effect of PSII herbicides, such as Irgarol, on corals and symbiotic dinoflagellates.

In vivo and in silico analyses of estrogenic potential of equine estrogens in medaka (Oryzias latipes).

Equine estrogens (EEs) are widely used in hormone replacement therapy pharmaceuticals for postmenopausal women. Previous studies have shown that EEs occur in the aquatic environment; however, the potential estrogenicity and risk of EEs in aquatic organisms, including fish, have yet to be studied in detail. Therefore, we evaluated the estrogenic potential of major EEs, namely equilin (Eq), 17α-dihydroequilin (17α-Eq), 17ß-dihydroequilin (17ß-Eq), equilenin (Eqn), 17α-dihydroequilenin (17α-Eqn), and 17ß-dihydroequilenin (17ß-Eqn), on medaka (Oryzias latipes) using in vivo and in silico assays. Quantitative real-time RT-PCR analyses revealed that expression levels of choriogenin L (ChgL) and choriogenin H (ChgH) in medaka embryos responded to various types and concentrations of EEs in a concentration-dependent manner, whereas transcription levels of vitellogenin 1 were not significantly affected by any of the EEs in the concentration range tested. The order of the in vivo estrogenic potencies of EEs was as follows: 17ß-Eq > Eq > 17ß-Eqn > Eqn > 17α-Eqn > 17α-Eq. Additionally, the 50% effective concentrations (EC50) of 17ß-Eq was lower than that of 17ß-estradiol. We also investigated the interaction potential of EEs with medaka estrogen receptor (ER) subtypes in silico using a three-dimensional model of the ligand-binding domain (LBD) for each ER and docking simulations. All six EEs were found to interact with the LBDs of ERα, ERß1, and ERß2. The order of the in silico interaction potentials of EEs with each ER LBD was as follows: 17ß-Eq > 17α-Eq > Eq > 17ß-Eqn > 17α-Eqn > Eqn. Furthermore, we identified the key amino acids that interact with EEs in each ER LBD; our findings suggest that amino acids and/or their hydrogen bonding may be responsible for the ligand-specific interactions with each ER. This study is the first to comprehensively analyze the estrogenic potential of EEs in medaka both in vivo and in silico.

Thermal tolerance of the hermatypic coral Acropora tenuis elucidated by RGB analysis and expression of heat shock proteins in coral and symbiotic dinoflagellates.

Increased seawater temperature has resulted in mass coral bleaching events globally. Acropora tenuis, the dominant hermatypic coral species in southern Japan, was exposed to four temperature treatments [28 °C, 30 °C, 32 °C, and >32 (=33.3 °C)] for 7 d. The coral colour was converted to R (red), G (green), and B (blue) values, each ranging from 0 (darkest) to 255 (brightest). RGB values exposed to 28 °C and 30 °C decreased slightly, whereas those exposed to 32 °C increased significantly after day 3-6, and those exposed to 33.3 °C changed to white within 2 d. Quantitative RT-PCR analysis revealed no significant changes in heat shock proteins in Acropora and symbiotic dinoflagellates at 28 °C and 30 °C after a 7 d exposure. Our findings revealed that 30 °C, higher than the mean temperature of the warmest month in southern Japan, was an inhabitable temperature for A. tenuis.

[Analysis and Health Risk Assessment of Cadmium and Arsenic in Japanese, Vietnamese, and Indonesian Rice].

Herein, we quantified the concentrations of cadmium (Cd) and arsenic (As) in 63 milled rice (Oryza sativa L.) cultivated in Japan, Vietnam, and Indonesia. We estimated the daily intake of Cd and As by adults and children consuming this rice by inductively coupled plasma-mass spectrometer (ICP-MS). Cd and As were detected in all milled rice samples. No significant differences were observed in Cd concentrations between Japanese (50th percentile concentration: 0.036 mg/kg), Vietnamese (0.035 mg/kg), and Indonesian rice (0.022 mg/kg). However, As concentrations in Vietnamese rice (50th percentile concentration: 0.142 mg/kg) were significantly higher than those in Japanese (0.101 mg/kg, p<0.001) and Indonesian rice (0.038 mg/kg, p<0.0001). Target hazard quotients (THQs) were then calculated to evaluate the non-carcinogenic health risk from ingestion of individual heavy metals (Cd and As) by rice consumption. Results revealed that THQs of individual heavy metals for Japanese, Vietnamese, and Indonesian adults and children consuming this rice were all less than one, suggesting that no health risk is associated with the intake of a single heavy metal via rice consumption.

Molecular Insights into Structural and Ligand Binding Features of Methoprene-Tolerant in Daphnids.

Juvenile hormone (JH) is an important endocrine factor regulating many biological activities in arthropods. In daphnids, methoprene-tolerant (Met) belongs to a basic helix-loop-helix/Per-Arnt-Sim (bHLH/PAS) family protein which has recently been confirmed as a JH receptor and can bind and be activated by JHs and JH agonists. Although the activation of the JH signaling pathway causes many physiological effects, the molecular basis for the structural feature and ligand binding properties of Daphnia Met are not fully understood. To study the ligand preference in terms of structural features of Daphnia Met, we built in silico homology models of the PAS-B domain of Daphnia Mets from cladoceran crustaceans, Daphnia pulex and D. magna. Structural comparison of two Daphnia Met PAS-B domain models revealed that the volume in the main cavity of D. magna Met was larger than that of D. pulex Met. Compared with insect Met, Daphnia Met had a less hydrophobic cavity due to polar residues in the core-binding site. Molecular docking simulations of JH and its analogs with Daphnia Met indicated that the interaction energies were correlated with each of the experimental values of in vivo JH activities based on male induction and in vitro Met-mediated transactivation potencies. Furthermore, in silico site-directed mutagenesis supported experimental findings that Thr292 in D. pulex Met and Thr296 in D. magna Met substitution to valine contribute to JH selectivity and differential species response. This study demonstrates that in silico simulations of Daphnia Met and its ligands may be a tool for predicting the ligand profile and cross species sensitivity.

Succession of delayed fluorescence correlated with coral bleaching in the hermatypic coral Acropora tenuis.

We investigated coral bleaching by monitoring colour changes and measuring the delayed fluorescence (DF) of symbiotic dinoflagellates in the hermatypic coral Acropora tenuis, exposed to 1.0 µg/L Irgarol 1051 (photosystem II herbicide) for 14 d. The Irgarol concentration corresponded to those from international port regions of the world. The coral colour and DFs under the control treatment were stable throughout the experiment, whereas under the Irgarol treatment the corals showed gradual bleaching. The Irgarol treatment caused a rapid decrease in the slow decay DF component (10.1-60.0 s), while the fast decay DF component (0.1-10.0 s) decreased significantly after 6 d. The significant correlation between the latter values and the coral colour indicates that if the electron accumulation function of quinones QA and QB is compromised, corals will bleach. The present study will contribute to the understanding of the mechanism involved in bleaching of coral exposed to herbicides.

Choriogenin transcription in medaka embryos and larvae as an alternative model for screening estrogenic endocrine-disrupting chemicals.

This study assessed the transcription levels of estrogen-responsive genes, such as vitellogenins (Vtg1 and Vtg2), choriogenins (ChgL, ChgH, and ChgHm), cytochrome P450 aromatase (CYP19a1b), and ER subtypes (ERα, ERß1, and ERß2), in 7 days-post-fertilization (dpf) embryos and 9 and 12 dpf larvae of medaka (Oryzias latipes) exposed to estrogenic endocrine-disrupting chemicals (EDCs). The <5 h-post-fertilization embryos were exposed to EDCs such as 17ß-estradiol (E2), p-n-nonylphenol (NP), and bisphenol A (BPA). In E2 (0.10-222 nM)-treated 7 dpf embryos and 9 or 12 dpf larvae, ChgL, ChgH, and ChgHm expression was up-regulated in a concentration-dependent manner. By contrast, interestingly, Vtg1 and Vtg2 expression was not induced in E2-treated 7 dpf embryos but was significantly induced in 9 and 12 dpf larvae, suggesting a developmental-stage-specific regulatory mechanism underlying Vtg expression. The maximum concentrations of NP (0.09-1.5 µM) and BPA (1.8-30 µM) up-regulated Chg expression in 9 or 12 dpf larvae, and the relative estrogenic potencies (REPs) of E2, NP, and BPA were 1, 2.1 × 10-4, and 1.0 × 10-5, respectively. Chg messenger RNA (mRNA) in medaka embryos and larvae can be used as a sensitive biomarker for screening potential estrogenic EDCs. Our assay system using embryos and larvae can be used as an in vivo alternative model because independent feeding stages (e.g., embryonic and early larval stages) are suitable alternatives.

Effects of ecologically relevant concentrations of Irgarol 1051 in tropical to subtropical coastal seawater on hermatypic coral Acropora tenuis and its symbiotic dinoflagellates.

The effects of ecologically relevant concentrations of Irgarol 1051, a representative PSII herbicide, on hermatypic corals were studied in the laboratory. The colour and chlorophyll fluorescence of Acropora tenuis were examined following exposure to around ambient concentrations of Irgarol 1051 (20 ng/L and 200 ng/L) for 7 days. While the colour of corals was stable throughout the experiment at both concentrations, the maximum effective quantum yield (ΔF/Fm') of symbiotic dinoflagellates decreased with increasing Irgarol 1051 concentration (day 7: 8%, 20 ng/L; 37%, 200 ng/L). The expression of heat shock protein (HSP) 70 and 90 in symbiotic dinoflagellates was upregulated after 7 days exposure to both Irgarol concentrations, whereas HSP90 in coral was not upregulated. The findings of the present study suggest that the threshold of chlorophyll fluorescence and HSP expression in symbiotic dinoflagellates is lower than 20 ng/L, which is around ecologically relevant concentrations in tropical to subtropical waters.

Potential mechanisms underlying embryonic developmental toxicity caused by benzo[a]pyrene in Japanese medaka (Oryzias latipes).

Polycyclic aromatic hydrocarbons (PAHs), such as benzo[a]pyrene (BaP), are widely distributed in air, water, and sediments; however, limited data are available regarding their potential adverse effects on the early life stages of fish. In this study, we evaluated the embryonic teratogenicity and developmental toxicity of BaP in Japanese medaka (Oryzias latipes) using a nanosecond pulsed electric field (nsPEF) technique and predicted their molecular mechanisms via transcriptome analysis. The gas chromatography/mass spectrometry analyses revealed that the BaP was efficiently incorporated into the embryos by nsPEF treatment. The embryos incorporating BaP presented typical teratogenic and developmental effects, such as cardiovascular abnormalities, developmental abnormalities, and curvature of backbone. DNA microarray analysis revealed several unique upregulated genes, such as those involved in cardiovascular diseases, various cellular processes, and neural development. Furthermore, the gene set enrichment and network analyses found several genes and hub proteins involved in the developmental effects of BaP on the embryos. These findings suggest a potential mechanism of teratogenicity and developmental toxicity caused by exposure to BaP. The nsPEF and transcriptome analyses in combination can be effective for evaluating the potential effects of chemical substances on medaka embryos.

In Vitro and In Silico Evaluations of Binding Affinities of Perfluoroalkyl Substances to Baikal Seal and Human Peroxisome Proliferator-Activated Receptor α.

In this study, we assessed the binding affinities of perfluoroalkyl substances (PFASs), including perfluoroalkyl carboxylates (PFCAs) and perfluoroalkyl sulfonates (PFSAs), to the ligand-binding domains (LBDs) of Baikal seal ( Pusa sibirica; bs) and human (h) peroxisome proliferator-activated receptor alpha (PPARα). An in vitro competitive binding assay showed that six PFCAs and two PFSAs could bind to recombinant bs and hPPARα LBD proteins in a dose-dependent manner. The relative binding affinities (RBAs) of PFASs to bsPPARα were as follows: PFOS > PFDA > PFNA > PFUnDA > PFOA > PFHxS > PFHpA > PFHxA. The RBAs to bsPPARα showed a significant positive correlation with those to hPPARα. In silico PPARα homology modeling predicted that there were two ligand-binding pockets (LBPs) in the bsPPARα and hPPARα LBDs. Structure-activity relationship analyses suggested that the binding potencies of PFASs to PPARα might depend on LBP binding cavity volume, hydrogen bond interactions, the number of perfluorinated carbons, and the hydrophobicity of PFASs. Interspecies comparison of the in vitro binding affinities revealed that bsPPARα had higher preference for PFASs with long carbon chains than hPPARα. The in silico docking simulations suggested that the first LBP of bsPPARα had higher affinities than that of hPPARα; however, the second LBP of bsPPARα had lower affinities than that of hPPARα. To our knowledge, this is the first evidence showing interspecies differences in the binding of PFASs to PPARαs and their structure-activity relationships.

In Vitro Assessment of Activation of Baikal Seal ( Pusa sibirica) Peroxisome Proliferator-Activated Receptor α by Polybrominated Diphenyl Ethers.

We investigated the Baikal seal ( Pusa sibirica) peroxisome proliferator-activated receptor α (bsPPARα) transactivation potencies of polybrominated diphenyl ethers (PBDEs) using an in vitro bsPPARα reporter gene assay. BDE47, BDE99, and BDE153 induced bsPPARα-mediated transcriptional activities in a dose-dependent manner. To compare bsPPARα transactivation potencies of PBDEs, perfluorooctanoic acid (PFOA)-based relative potencies (REPs), a ratio of 50% effective concentration of PFOA to the test chemical, were determined. The order of REPs of PBDEs was BDE153 (13) > BDE99 (8.1) > BDE47 (6.6) > PFOA (1.0) > BDE100, BDE154, and BDE183 (not activated). PBDEs with two bromine atoms at the ortho position showed higher bsPPARα transactivation potencies than those with three bromine atoms. Comparison of the lowest-observed-effect concentration in bsPPARα reporter gene assays revealed that BDE99 was 7-fold more potent than CB99, a polychlorinated biphenyl congener with the same IUPAC number, indicating that brominated congeners could more efficiently activate bsPPARα than chlorinated congeners. The REPs of PBDEs for bsPPARα transactivation were approximately 7- to 13-fold higher than those of perfluorochemicals (PFCs), suggesting that the effects of PBDEs on the bsPPARα signaling pathway may be superior to those of PFCs. This study provides the first evidence that PBDE congeners activate PPARα in vitro.

Identification and characterization of heat shock protein 90 (HSP90) in the hard coral Acropora tenuis in response to Irgarol 1051.

To elucidate the effects of the herbicide Irgarol 1051 on the gene expression of heat shock protein 90 (HSP90) in hard corals, we isolated a full-length cDNA encoding HSP90 from Acropora tenuis, which has a deduced open reading frame of 732-amino acid residues with a predicted molecular mass of 84.5 kDa. The amino acid sequence of A. tenuis HSP90 showed a high degree of similarity with the hermatypic-coral HSP90 families. After a 7-d exposure to 1 or 10 µg/L of Irgarol, the body colours of corals in the 10 µg/L treatment group were significantly whiter (bleached), whereas no such effects were observed in the corals in the 1 µg/L treatment group. However, the expression level of coral HSP90 was significantly downregulated after exposure to both 1 and 10 µg/L Irgarol. These results suggest that A. tenuis HSP90 may be a useful molecular biomarker to predict bleaching caused by herbicides.