Cytochrome P450 family 1 (CYP1) proteins are important in a large number of toxicological processes. CYP1A and CYP1B genes are well known in mammals, but the evolutionary history of the CYP1 family as a whole is obscure; that history may provide insight into endogenous functions of CYP1 enzymes. Here, we identify CYP1-like genes in early deuterostomes (tunicates and echinoderms), and several new CYP1 genes in vertebrates (chicken, Gallus gallus and frog, Xenopus tropicalis). Profile hidden Markov models (HMMs) generated from vertebrate CYP1A and CYP1B protein sequences were used to identify 5 potential CYP1 homologs in the tunicate Ciona intestinalis genome. The C. intestinalis genes were cloned and sequenced, confirming the predicted sequences. Orthologs of 4 of these genes were found in the Ciona savignyi genome. Bayesian phylogenetic analyses group the tunicate genes in the CYP1 family, provisionally in 2 new subfamilies, CYP1E and CYP1F, which fall in the CYP1A and CYP1B/1C clades. Bayesian and maximum likelihood analyses predict functional divergence between the tunicate and vertebrate CYP1s, and regions within CYP substrate recognition sites were found to differ significantly in position-specific substitution rates between tunicates and vertebrates. Subsequently, 10 CYP1-like genes were found in the echinoderm Strongylocentrotus purpuratus (sea urchin) genome. Several of the tunicate and echinoderm CYP1-like genes are expressed during development. Canonical xenobiotic response elements are present in the upstream genomic sequences of most tunicate and sea urchin CYP1s, and both groups are predicted to possess an aryl hydrocarbon receptor (AHR), suggesting possible regulatory linkage of AHR and these CYPs. The CYP1 family has undergone multiple rounds of gene duplication followed by functional divergence, with at least one gene lost in mammals. This study provides new insight into the origin and evolution of CYP1 genes.
Receiver operating characteristic (ROC) curve analysis is a simple and effective means to compare the accuracies of indicator variables of bacterial beach water quality. The indicator variables examined in this study were previous day's Enterococcus density and antecedent rainfall at 24, 48, and 96 h. Daily Enterococcus densities and 15-min rainfall values were collected during a 5-year (1996 to 2000) study of four Boston Harbor beaches. The indicator variables were assessed for their ability to correctly classify water as suitable or unsuitable for swimming at a maximum threshold Enterococcus density of 104 CFU/100 ml. Sensitivity and specificity values were determined for each unique previous day's Enterococcus density and antecedent rainfall volume and used to construct ROC curves. The area under the ROC curve was used to compare the accuracies of the indicator variables. Twenty-four-hour antecedent rainfall classified elevated Enterococcus densities more accurately than previous day's Enterococcus density (P ؍ 0.079). An empirically derived threshold for 48-h antecedent rainfall, corresponding to a sensitivity of 0.75, was determined from the 1996 to 2000 data and evaluated to ascertain if the threshold would produce a 0.75 sensitivity with independent water quality data collected in 2001 from the same beaches.
Background Sterol 14α-demethylase (cytochrome P450 51, CYP51, P45014DM) is a microsomal enzyme that in eukaryotes catalyzes formation of sterols essential for cell membrane function and as precursors in biosynthesis of steroid hormones. Functional properties of CYP51s are unknown in non-mammalian deuterostomes. Methods PCR-cloning and sequencing and computational analyses (homology modeling and docking) addressed CYP51 in zebrafish Danio rerio, the reef fish sergeant major Abudefduf saxatilis, and the sea urchin Strongylocentrotus purpuratus. Following N-terminal amino acid modification, zebrafish CYP51 was expressed in Escherichia coli, and lanosterol 14α-demethylase activity and azole inhibition of CYP51 activity were characterized using GC/MS. Results Molecular phylogeny positioned S. purpuratus CYP51 at the base of the deuterostome clade. In zebrafish, CYP51 is expressed in all organs examined, most strongly in intestine. The recombinant protein bound lanosterol and catalyzed 14α-demethylase activity, at 3.2 nmol/min/nmol CYP51. The binding of azoles to zebrafish CYP51 gave KS (dissociation constant) values of 0.26 μM for ketoconazole and 0.64 μM for propiconazole. Displacement of carbon monoxide also indicated zebrafish CYP51 has greater affinity for ketoconazole. Docking to homology models showed that lanosterol docks in fish and sea urchin CYP51s with an orientation essentially the same as in mammalian CYP51. Docking of ketoconazole indicates it would inhibit fish and sea urchin CYP51s. Conclusions Biochemical and computational analyses are consistent with lanosterol being a substrate for early deuterostome CYP51s. General Significance The results expand the phylogenetic view of animal CYP51, with evolutionary, environmental and therapeutic implications.
This review covers selected 2014 articles on the biological effects of pollutants and human physical disturbances on marine and estuarine plants, animals, ecosystems and habitats. The review, based largely on journal articles, covers field and laboratory measurement activities (bioaccumulation of contaminants, field assessment surveys, toxicity testing and biomarkers) as well as pollution issues of current interest including endocrine disrupters, emerging contaminants, wastewater discharges, dredging and disposal, etc. Special emphasis is placed on effects of oil spills and marine debris due in part to the 2010 Deepwater Horizon oil blowout in the Gulf of Mexico and the 2011 Japanese tsunami. Several topical areas reviewed in the past (ballast water and ocean acidification) were dropped this year. The focus of this review is on effects, not pollutant fate and transport. There is considerable overlap across subject areas (e.g.some bioaccumulation papers may be cited in other topical categories). Please use keyword searching of the text to locate related but distributed papers. Use this review only as a guide and please consult the original papers before citing them.
This review covers selected 2019 articles on the biological effects of pollutants, including human physical disturbances, on marine and estuarine plants, animals, ecosystems, and habitats. The review, based largely on journal articles, covers field, and laboratory measurement activities (bioaccumulation of contaminants, field assessment surveys, toxicity testing, and biomarkers) as well as pollution issues of current interest including endocrine disrupters, emerging contaminants, wastewater discharges, marine debris, dredging, and disposal. Special emphasis is placed on effects of oil spills and marine debris due largely to the 2010 Deepwater Horizon oil blowout in the Gulf of Mexico and proliferation of data on the assimilation and effects of marine debris microparticulates. Several topical areas reviewed in the past (e.g., mass mortalities ocean acidification) were dropped this year. The focus of this review is on effects, not on pollutant sources, chemistry, fate, or transport. There is considerable overlap across subject areas (e.g., some bioaccumulation data may be appeared in other topical categories such as effects of wastewater discharges, or biomarker studies appearing in oil toxicity literature). Therefore, we strongly urge readers to use keyword searching of the text and references to locate related but distributed information. Although nearly 400 papers are cited, these now represent a fraction of the literature on these subjects. Use this review mainly as a starting point. And please consult the original papers before citing them.
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