Environmental concentrations of the selective serotonin reuptake inhibitor fluoxetine impact specific behaviors involved in reproduction, feeding and predator avoidance in the fish Pimephales promelas (fathead minnow)

AbstractPharmaceuticals and personal care products (PPCPs) have been found in surface waters worldwide, but little is understood of their effects on the wildlife that inhabit these waters. Fluoxetine (Prozac; Eli Lilly), a highly prescribed selective serotonin reuptake inhibitor (SSRI), is a commonly found PPCP in surface water. The purpose of this project was to determine if environmentally relevant concentrations of fluoxetine impact behavior that is important for population survival in native fish species, including reproduction, feeding and predator avoidance. Chronic 4-week exposures were conducted with doses ranging from 100ng/L to 100μg/L to cover a range of environmentally relevant concentrations up to higher concentrations comparable to other published studies with the same drug that have documented various physiological impacts. Pimephales promelas (fathead minnow), a species native to North America, was used as it conducts a range of specific mating behaviors and therefore serves as an excellent model of specific impacts on brain function. Fluoxetine concentrations as low as 1μg/L, a concentration that has been found in many freshwater environments, were found to significantly impact mating behavior, specifically nest building and defending in male fish. Males were also found to display aggression, isolation, and repetitive behaviors at higher concentrations. Female mating behavior was largely unaffected. In addition, predator avoidance behaviors in males and females were also impacted at 1μg/L. Feeding was impacted at 10μg/L and in the highest exposure (100μg/L), egg production was limited by deaths of females due to significant male aggressive behaviors in the first two weeks of exposure. Specific behavioral changes occurred at each concentration (most noticeably 1μg/L and 100μg/L) indicating a dose dependent effect that triggered different responses at lower exposures versus higher exposures or differential impacts of dose depending on brain region. Length of exposure also had an impact on aggressive behavior. Changes in hormone levels, indicating significant neuroendocrine changes, suggested as a mechanism of response in higher dose and acute studies, were not linked to changes in behaviors at the doses used in this study. This research provides detailed data on how exposures to fluoxetine impact specific fish behaviors and reproduction and that the effects are dose dependent

Monitoring Protein Splicing Using In-gel Fluorescence Immediately Following SDS-PAGE

Inteins garner significant interest from both basic and applied researchers due to their unique catalytic abilities. Herein, we describe a protocol for accurately monitoring protein splicing without purification using in-gel fluorescence immediately following Tris-Glycine SDS-PAGE. Following expression in Escherichia coli, cells are lysed by sonication, cell supernatants are separated using Tris- Glycine SDS-PAGE, and superfolder GFP (sfGFP) fluorescence is directly visualized within gels. This method is rapid, with sfGFP immediately imaged following SDS-PAGE without staining. Further, sfGFP can be specifically detected in complex samples such as E. coli cell supernatants, proteins run at expected masses, and all steps can be performed at ambient temperature. This strategy is broadly applicable beyond the study of protein splicing and can be used for sensitive and specific visualization of superfolder sfGFP-tagged proteins in-gel

Reactive Chlorine Species Reversibly Inhibit DnaB Protein Splicing in Mycobacteria

Intervening proteins, or inteins, are mobile genetic elements that are translated within host polypeptides and removed at the protein level by splicing. In protein splicing, a self-mediated reaction removes the intein, leaving a peptide bond in place. While protein splicing can proceed in the absence of external cofactors, several examples of conditional protein splicing (CPS) have emerged. In CPS, the rate and accuracy of splicing are highly dependent on environmental conditions. Because the activity of the intein-containing host protein is compromised prior to splicing and inteins are highly abundant in the microbial world, CPS represents an emerging form of posttranslational regulation that is potentially widespread in microbes. Reactive chlorine species (RCS) are highly potent oxidants encountered by bacteria in a variety of natural environments, including within cells of the mammalian innate immune system. Here, we demonstrate that two naturally occurring RCS, namely, hypochlorous acid (the active compound in bleach) and N-chlorotaurine, can reversibly block splicing of DnaB inteins from Mycobacterium leprae and Mycobacterium smegmatis in vitro. Further, using a reporter that monitors DnaB intein activity within M. smegmatis, we show that DnaB protein splicing is inhibited by RCS in the native host. DnaB, an essential replicative helicase, is the most common intein-housing protein in bacteria. These results add to the growing list of environmental conditions that are relevant to the survival of the intein-containing host and influence protein splicing, as well as suggesting a novel mycobacterial response to RCS. We propose a model in which DnaB splicing, and therefore replication, is paused when these mycobacteria encounter RCS

Imaging the Molecular Disk Orbiting the Twin Young Suns of V4046 Sgr

We have imaged the disk surrounding the nearby (D~73 pc), ~12 Myr, classical T Tauri binary system V4046 Sgr with the Submillimeter Array (SMA) at an angular resolution of ~2". We detect a rotating disk in 12CO(2-1) and 13CO(2-1) emission, and resolve the continuum emission at 1.3 mm. We infer disk gas and dust masses of ~110 and ~40 Earth masses, respectively. Fits to a power-law disk model indicate that the molecular disk extends to ~370 AU and is viewed at an inclination of between ~33 and ~39 degrees for dynamical stellar masses ranging from 1.8 $M_\odot$ down to 1.5 $M_\odot$ (the range of total mass previously determined for the central, 2.4 day spectroscopic binary). This range of disk inclination is consistent with that assumed in deducing the central binary mass (i.e., 35 degrees), suggesting that the V4046 Sgr binary system and its circumbinary, molecular disk are coplanar. In light of the system's age and binarity, the presence of an extensive molecular disk orbiting V4046 Sgr provides constraints on the timescales of processes related to Jovian planet formation, and demonstrates that circumbinary Jovian planets potentially could form around close binary systems.Comment: 17 pages, 8 figures, accepted for publication in Ap

Variants in the estrogen receptor alpha gene and its mRNA contribute to risk for schizophrenia

Estrogen modifies human emotion and cognition and impacts symptoms of schizophrenia. We hypothesized that the variation in the estrogen receptor alpha (ESR1) gene and cortical ESR1 mRNA is associated with schizophrenia. In a small case–control genetic association analysis of postmortem brain tissue, genotype CC (rs2234693) and haplotypes containing the C allele of a single-nucleotide polymorphism (SNP) in intron1 (PvuII) were more frequent in African American schizophrenics (P = 0.01–0.001). In a follow-up family-based association analysis, we found overtransmission of PvuII allele C and a PvuII C-containing haplotype (P = 0.01–0.03) to African American and Caucasian patients with schizophrenia. Schizophrenics with the ‘at risk’ PvuII genotype had lower ESR1 mRNA levels in the frontal cortex. Eighteen ESR1 splice variants and decreased frequencies of the wild-type ESR1 mRNA were detected in schizophrenia. In one patient, a unique ESR1 transcript with a genomic insert encoding a premature stop codon and a truncated ESR1 protein lacking most of the estrogen binding domain was the only transcript detected. Using a luciferase assay, we found that mRNA encoding a truncated ESR1 significantly attenuates gene expression at estrogen-response elements demonstrating a dominant negative function. An intron 6 SNP [rs2273207(G)] was associated with an ESR1 splice variant missing exon seven. The T allele of another intron 6 SNP was part of a 3′ haplotype less common in schizophrenia [rs2273206(T), rs2273207(G), rs2228480(G)]. Thus, the variation in the ESR1 gene is associated with schizophrenia and the mechanism of this association may involve alternative gene regulation and transcript processing

Evaluating the End-User Experience of Private Browsing Mode

Nowadays, all major web browsers have a private browsing mode. However, the mode's benefits and limitations are not particularly understood. Through the use of survey studies, prior work has found that most users are either unaware of private browsing or do not use it. Further, those who do use private browsing generally have misconceptions about what protection it provides. However, prior work has not investigated \emph{why} users misunderstand the benefits and limitations of private browsing. In this work, we do so by designing and conducting a three-part study: (1) an analytical approach combining cognitive walkthrough and heuristic evaluation to inspect the user interface of private mode in different browsers; (2) a qualitative, interview-based study to explore users' mental models of private browsing and its security goals; (3) a participatory design study to investigate why existing browser disclosures, the in-browser explanations of private browsing mode, do not communicate the security goals of private browsing to users. Participants critiqued the browser disclosures of three web browsers: Brave, Firefox, and Google Chrome, and then designed new ones. We find that the user interface of private mode in different web browsers violates several well-established design guidelines and heuristics. Further, most participants had incorrect mental models of private browsing, influencing their understanding and usage of private mode. Additionally, we find that existing browser disclosures are not only vague, but also misleading. None of the three studied browser disclosures communicates or explains the primary security goal of private browsing. Drawing from the results of our user study, we extract a set of design recommendations that we encourage browser designers to validate, in order to design more effective and informative browser disclosures related to private mode

Treatment Use and Costs among Privately-Insured Youth with Bipolar Diagnoses

Recent evidence suggests that children are increasingly diagnosed with bipolar disorder, yet no studies have quantified treatment costs for pediatric patients. The objectives are to identify one-year health services utilization and treatment costs among youth with new bipolar diagnoses

Projected temperature-related deaths in ten large U.S. metropolitan areas under different climate change scenarios.

BACKGROUND: There is an established U-shaped association between daily temperature and mortality. Temperature changes projected through the end of century are expected to lead to higher rates of heat-related mortality but also lower rates of cold-related mortality, such that the net change in temperature-related mortality will depend on location. OBJECTIVES: We quantified the change in heat-, cold-, and temperature-related mortality rates through the end of the century across 10 large US metropolitan areas. METHODS: We applied location-specific projections of future temperature from over 40 downscaled climate models to exposure-response functions relating daily temperature and mortality in 10 US metropolitan areas to estimate the change in temperature-related mortality rates in 2045-2055 and 2085-2095 compared to 1992-2002, under two greenhouse gas emissions scenarios (RCP 4.5 and 8.5). We further calculated the total number of deaths attributable to temperature in 1997, 2050, and 2090 in each metropolitan area, either assuming constant population or accounting for projected population growth. RESULTS: In each of the 10 metropolitan areas, projected future temperatures were associated with lower rates of cold-related deaths and higher rates of heat-related deaths. Under the higher-emission RCP 8.5 scenario, 8 of the 10 metropolitan areas are projected to experience a net increase in annual temperature-related deaths per million people by 2086-2095, ranging from a net increase of 627 (95% empirical confidence interval [eCI]: 239, 1018) deaths per million in Los Angeles to a net decrease of 59 (95% eCI: -485, 314) deaths per million in Boston. Applying these projected temperature-related mortality rates to projected population size underscores the large public health burden of temperature. CONCLUSIONS: Increases in the heat-related death rate are projected to outweigh decreases in the cold-related death rate in 8 out of 10 cities studied under a high emissions scenario. Adhering to a lower greenhouse gas emissions scenario has the potential to substantially reduce future temperature-related mortality

Comparison of quantitative trait loci methods:Total expression and allelic imbalance method in brain RNA-seq

BackgroundOf the 108 Schizophrenia (SZ) risk-loci discovered through genome-wide association studies (GWAS), 96 are not altering the sequence of any protein. Evidence linking non-coding risk-SNPs and genes may be established using expression quantitative trait loci (eQTL). However, other approaches such allelic expression quantitative trait loci (aeQTL) also may be of use.MethodsWe applied both the eQTL and aeQTL analysis to a biobank of deeply sequenced RNA from 680 dorso-lateral pre-frontal cortex (DLPFC) samples. For each of 340 genes proximal to the SZ risk-SNPs, we asked how much SNP-genotype affected total expression (eQTL), as well as how much the expression ratio between the two alleles differed from 1:1 as a consequence of the risk-SNP genotype (aeQTL).ResultsWe analyzed overlap with comparable eQTL-findings: 16 of the 30 risk-SNPs known to have gene-level eQTL also had gene-level aeQTL effects. 6 of 21 risk-SNPs with known splice-eQTL had exon-aeQTL effects. 12 novel potential risk genes were identified with the aeQTL approach, while 55 tested SNP-pairs were found as eQTL but not aeQTL. Of the tested 108 loci we could find at least one gene to be associated with 21 of the risk-SNPs using gene-level aeQTL, and with an additional 18 risk-SNPs using exon-level aeQTL.ConclusionOur results suggest that the aeQTL strategy complements the eQTL approach to susceptibility gene identification