Removal of ecotoxicity of 17α-ethinylestradiol using TAML/peroxide water treatment

17α -ethinylestradiol (EE2), a synthetic oestrogen in oral contraceptives, is one of many pharmaceuticals found in inland waterways worldwide as a result of human consumption and excretion into wastewater treatment systems. At low parts per trillion (ppt), EE2 induces feminisation of male fish, diminishing reproductive success and causing fish population collapse. Intended water quality standards for EE2 set a much needed global precedent. Ozone and activated carbon provide effective wastewater treatments, but their energy intensities and capital/operating costs are formidable barriers to adoption. Here we describe the technical and environmental performance of a fast- developing contender for mitigation of EE2 contamination of wastewater based upon smallmolecule, full-functional peroxidase enzyme replicas called “TAML activators”. From neutral to basic pH, TAML activators with H2O2 efficiently degrade EE2 in pure lab water, municipal effluents and EE2-spiked synthetic urine. TAML/H2O2 treatment curtails estrogenicity in vitro and substantially diminishes fish feminization in vivo. Our results provide a starting point for a future process in which tens of thousands of tonnes of wastewater could be treated per kilogram of catalyst. We suggest TAML/H2O2 is a worthy candidate for exploration as an environmentally compatible, versatile, method for removing EE2 and other pharmaceuticals from municipal wastewaters.Heinz Endowments, the Swiss National Science Foundation, the Steinbrenner Institute for a Steinbrenner Doctoral Fellowship. NMR instrumentation at CMU was partially supported by NSF (CHE-0130903 and CHE-1039870)

Levels of State and Trait Anxiety in Patients Referred to Ophthalmology by Primary Care Clinicians: A Cross Sectional Study

Purpose There is a high level of over-referral from primary eye care leading to significant numbers of people without ocular pathology (false positives) being referred to secondary eye care. The present study used a psychometric instrument to determine whether there is a psychological burden on patients due to referral to secondary eye care, and used Rasch analysis to convert the data from an ordinal to an interval scale. Design Cross sectional study. Participants and Controls 322 participants and 80 control participants. Methods State (i.e. current) and trait (i.e. propensity to) anxiety were measured in a group of patients referred to a hospital eye department in the UK and in a control group who have had a sight test but were not referred. Response category analysis plus infit and outfit Rasch statistics and person separation indices were used to determine the usefulness of individual items and the response categories. Principal components analysis was used to determine dimensionality. Main Outcome Measure Levels of state and trait anxiety measured using the State-Trait Anxiety Inventory. Results State anxiety scores were significantly higher in the patients referred to secondary eye care than the controls (p0.1). Rasch analysis highlighted that the questionnaire results needed to be split into “anxiety-absent” and “anxiety-present” items for both state and trait anxiety, but both subscales showed the same profile of results between patients and controls. Conclusions State anxiety was shown to be higher in patients referred to secondary eye care than the controls, and at similar levels to people with moderate to high perceived susceptibility to breast cancer. This suggests that referral from primary to secondary eye care can result in a significant psychological burden on some patients

Benthic Macroinvertebrate Community as an Indicator of Stream Health: The Effects of Land Use on Stream Benthic Macroinvertebrates

Biomonitoring of stream health in the tropics still emphasize on the use of standard water chemistry methods (physicochemical variables), which require expensive and elaborate measuring apparatus. In this study, the reliability of benthic macroinvertebrates as bioindicators of freshwaterstreams was carried out. The study also attempted to determine thediscriminating power of various biotic indices in characterizing sitesacross land use. Benthic macroinvertebrate samples were obtained fromnine streams in Silago, Southern Leyte and were identified to familylevel. One-way analysis of variance was performed on various bioticindices to assess the water quality of streams based on land use. AverageTolerance Score per Taxon (ATSPT) was the only biotic index thatdifferentiated the nine streams based on land use (P<0.001). Forestedsites achieved the lowest ATSPT score, whereas mixed forested-agriculturalsites had the highest ATSPT scores. Physicochemicalvariables (e.g., stream width, conductivity, total dissolved solids, watertemperature) and biological metrics (e.g., Simpson’s diversity index, totalmacroinvertebrate density) used in the study supported this assessment.The results show that benthic macroinvertebrates can be used aspotential biomonitoring tool to evaluate the ecological integrity ofwaterways in the country. Long-term data sets will be generated fromfuture sampling efforts for the development of the Philippine BioticIndex.Keywords: Average Tolerance Score per Taxon (ATSPT), biotic indices,stream monitoring, physicochemical, Philippine

Benthic Macroinvertebrates of the University of the Philippines Diliman Campus Waterways and Their Variation Across Land Use in an Urban, Academic Landscape

Urban development impacts stream ecosystems primarily via changes in hydrological regime, geomorphology, and in water quality. These changes in turn have biological effects. The University of the Philippines Diliman campus, located at the heart of the highly urbanized Quezon City, has gone through numerous developments in terms of landscape and infrastructure. Unlike the terrestrial environment, the extent to which these developments have impacted the campus waterways is unknown. Hence, our research aims to assess the overall condition of the waterways in the campus based on the benthic macroinvertebrate assemblages. A total of 19 stream reaches were sampled in November 2015 and 2016 in the following land use categories: academic/academic support units (six sites), campus core (eight sites), and parks and open spaces (five sites). One-way analysis of variance (ANOVA) detected significant spatial difference in several macroinvertebratebased metrics, stream physicochemistry, and in-stream habitat condition elements. Our study reveals that all sampled stream reaches, regardless of their land use categories, are under poor to severe pollution conditions. All macroinvertebrate-based metrics and indices indicate degraded water quality and stream health. Our results are consistent with urban stream studies elsewhere, which suggest that land-based activities can be stressful for some aquatic organisms, and at times, result in reduced abundance and even reduction in species composition.Keywords: Biomonitoring, biotic indices, stream habitat assessment, urban land use, water qualit

Genetics of cold-adapted B/Ann Arbor/1/66 influenza virus reassortants: the acidic polymerase (PA) protein gene confers temperature sensitivity and attenuated virulence

The cold-adapted B/Ann Arbor/1/66 influenza virus (ca B/AA/1/66) expresses temperature-sensitive (ts), cold-adapted (ca) and attenuation phenotypes. Reassortants which inherit one or more genes from ca B/AA/1/66 and all other genes from a virulent, wild-type influenza virus, B/Houston/1732/76, were produced and evaluated in order to identify the gene(s) responsible for the ts, ca and attenuation phenotypes. Only reassortants which inherited the PA gene from ca B/AA/1/66 expressed the ts phenotype in MDCK cells at 39 [deg]C. None of the reassortants tested expressed the ca phenotype in embryonated eggs at 25 [deg]C. The virulence of several reassortants was evaluated in ferrets. Inheritance of the PA gene from ca B/AA/1/66 was correlated with significant febrile attentuation and the apparent restriction of viral replication in the lower respiratory tract. Isolation of a virulent, non-ts revertant virus inheriting only the PA gene from ca B/AA/1/66 established a direct relationship between expression of the ts phenotype and attenuated virulence. Evidence for the contribution of at least one other gene from ca B/AA/1/66 to attenuation was observed. Thus, based on the methods used to determine reassortant gene compositions, these results indicate that the PA gene is primarily responsible for attenuation of ca B/AA/1/66 and its reassortants.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/26616/1/0000157.pd

Toward a typology of river functioning: a comprehensive study of the particulate organic matter composition at the multi-river scale

In river systems, particulate organic matter (POM) originates from various sources with distinct dynamics related to production, decomposition, transport, and burial, leading to spatiotemporal heterogeneity in the POM pool. This study uses C and N isotope and element ratios, Bayesian models and multivariate analyses to (1) quantify relationships between POM composition and environmental forcings, and (2) propose a typology of river functioning based on POM composition and their seasonal dynamics. Twenty-three temperate rivers, representing a large diversity of temperate conditions, were sampled fortnightly to monthly over one to seven years at their River-Estuary Interface (REI). Phytoplankton and labile terrestrial material were found in all rivers, while sewage and refractory terrestrial material appeared in a few. Across all rivers, phytoplankton dominance correlated with agricultural surfaces, labile terrestrial material with organic-rich leached soil, and refractory terrestrial matter with steep, soil-poor catchments. Seasonal dynamics mainly depended to phytoplankton growth, river discharge, and sediment resuspension. Statistical regionalisation identified four river-dynamics types: (1) POM is dominated by labile terrestrial material year-round; (2) POM is composed of labile and refractory terrestrial material plus phytoplankton with variable seasonality; (3) POM is composed of phytoplankton and labile terrestrial material without pronounced seasonality; and (4) POM is composed of phytoplankton and labile terrestrial material with pronounced seasonality. This research offers a comprehensive understanding of POM composition, dynamics, and drivers at the REI in temperate climates, complementing similar study in coastal systems. Future similar research on estuaries is called to refine knowledge along the Land-Ocean Aquatic Continuum.</p

Resolution of a common RNA sequencing ambiguity by terminal deoxynucleotidyl transferase

One of the more common ambiguities which arise when using reverse transcriptase and dideoxynucleotide-chain termination to sequence RNA is a radioactive band of cDNA that extends over all four lanes on a sequencing gel. The adjacent sequences both above and below the band are not affected. Assuming then, that these ambiguities are caused by the termination of the DNA polymerase activity of reverse transcriptase for reasons other than the insertion of a dideoxynucleotide in the growing cDNA chain, terminal deoxynucleotidyl transferase should be able to continue to add deoxynucleotides to these products after the sequencing reaction is complete. It does, clearing the improperly terminated cDNA from these pileup sites, revealing the correct sequence. This technique can also be used to identify the template RNA's 5'-terminal base, although far more units of terminal deoxynucleotidyl transferase are required.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/26058/1/0000132.pd

Sequence comparison of wild-type and cold-adapted B/Ann Arbor/1/66 influenza virus genes

Consensus sequences for both wt and ca B/Ann Arbor/1 /66 viral PB2, PB1, PA, NP, M, and NS genes were directly determined from vRNA using a combination of chemical and chain-termination sequencing methods. There were 105 sites of difference between the wt and ca sets of these six RNA genes. The differences resulted in 26 amino acid substitutions distributed over the six proteins. The sequence changes were compared to the sequences of other known influenza type B wt viruses to pinpoint those changes that were unique to the ca B/Ann Arbor/1/66 virus. Of the 26 amino acid differences, only 11 were unique to the cold-adapted virus. These unique sites were distributed among five of the six genes. The NS protein had no amino acid substitutions. The sequence changes are discussed in terms of their probable mode of origin and selection, and in terms of their importance to the cold-adapted, temperature-sensitive, and attenuation phenotypes of ca B/AA/1 /66 virus. The sequence and organization of the PB2 gene and predicted protein are also given. The PB2 gene was 2396 nucleotides long, and it encoded a predicted protein of 770 amino acids with a molecular weight of 88,035 Da for the wt virus and 88,072 Da for the ca virus. Both proteins were predominantly hydrophilic, and each had an overall charge of +24.5 at pH 7.0.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/27362/1/0000387.pd

A mutation in the PA protein gene of cold-adapted B/Ann Arbor/1/66 influenza virus associated with reversion of temperature sensitivity and attenuated virulence

Reassortant SG3 inherits only the acidic polymerase (PA) protein gene from the cold-adapted B/AA/1/66 influenza virus (ca B/AA/l/66) and all remaining genes from a virulent, wild-type virus. This reassortant demonstrates attenuated virulence in ferrets and expresses a is phenotype characteristic of the ca parent. During virulence evaluation of SG3, a virulent, non-ts revertant virus (designated SG3rFL) was isolated from the lungs of one ferret. In order to determine whether the reversion of SG3 resulted from mutation of the PA gene and/or as the result of extragenic supressor mutations, the revertant PA gene of SG3rFL was transferred to a reassortant (SG3r) inheriting only the revertant PA gene from SG3rFL and all remaining genes from SG3. Reassortant SG3r was non-ts and virulent, indicating that mutation of the PA gene was sufficient for the reversion of the is and attenuation phenotypes expressed by SG3rFL. The nucleotide and predicted amino acid sequences of the SG3rFL PA gene were determined and compared to those of wt and ca B/AA/1 /66. The predicted PA proteins of wt and ca B/AA/1 /66 are known to differ by six amino acid substitutions including a valine to methionine substitution at residue 431. The PA proteins of ca B/AA/1/66 and SG3rFL were distinguished by only the single amino acid substitution of methionine to isoluecine also occurring at residue 431. Thus, the methionine residue was implicated in the attenuation of ca B/AA/1/66 and its reassortants. The hydropathic properties of valine, isoleucine, and methionine suggested that reversion involved the restoration of hydrophobic character at this site.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/27363/1/0000388.pd

An inter-laboratory comparison demonstrates that [1H]-NMR metabolite fingerprinting is a robust technique for collaborative plant metabolomic data collection

In any metabolomics experiment, robustness and reproducibility of data collection is of vital importance. These become more important in collaborative studies where data is to be collected on multiple instruments. With minimisation of variance in sample preparation and instrument performance it is possible to elucidate even subtle differences in metabolite fingerprints due to genotype or biological treatment. In this paper we report on an inter laboratory comparison of plant derived samples by [1H]-NMR spectroscopy across five different sites and within those sites utilising instruments with different probes and magnetic field strengths of 9.4 T (400 MHz), 11.7 T (500 MHz) and 14.1 T (600 MHz). Whilst the focus of the study is on consistent data collection across laboratories, aspects of sample stability and the requirement for sample rotation within the NMR magnet are also discussed. Comparability of the datasets from participating laboratories was exceptionally good and the data were amenable to comparative analysis by multivariate statistics. Field strength differences can be adjusted for in the data pre-processing and multivariate analysis demonstrating that [1H]-NMR fingerprinting is the ideal technique for large scale plant metabolomics data collection requiring the participation of multiple laboratories