A method for the determination of genetic sex in the fathead minnow, Pimephales promelas, to support testing of endocrine-active chemicals

Certain endocrine-active toxicants have been reported to completely sex reverse both male and female individuals in amphibian, avian, fish, invertebrate, and reptile species, resulting in a phenotype indistinguishable from unaffected individuals. Detection of low-level sex reversal often requires large numbers of organisms to achieve the necessary statistical power, especially in those species with predominantly genetic sex determination and cryptic/homomorphic sex chromosomes. Here we describe a method for determining the genetic sex in the commonly used ecotoxicological model, the fathead minnow (Pimephales promelas). Analysis of amplified fragment length polymorphisms (AFLP) in a spawn of minnows resulted in detection of 10 sex-linked AFLPs, which were isolated and sequenced. No recombination events were observed with any sex-linked AFLP in the animals examined (n=112). A polymerase chain reaction (PCR) method was then developed that determined the presence of one of these sex-linked polymorphisms for utilization in routine toxicological testing. Analyses of additional spawns from our in-house culture indicate that fathead minnows utilize a XY sex determination strategy and confirm that these markers can be used to genotype sex; however, this method is currently limited to use in laboratory studies in which breeders possess a defined genetic makeup. The genotyping method described herein can be incorporated into endocrine toxicity assays that examine the effects of chemicals on gonad differentiation.     

Toward a relational politics of representation

Rhetorical theories of representation, caught in the logics of transcendence/immanence, have struggled to reconcile the need to move beyond representation with the political importance of critiquing representational effects. I argue that this tension can—and must—be addressed through a relational politics of representation that draws from antiracist and decolonial theory. Tracing poststructural critiques of representational ontology, epistemology, and politics, I demonstrate their dependence on racializing and colonial processes. I then describe how rethinking our theories of representation relationally figures both ontology and epistemology as inherently political, and opens the possibility for theorizing the human beyond Man. I argue that a relational politics of representation is an impossible necessity that must be continually (re)attempted though it will never be fully achieved.     

Determining Ethyl Esters in Fish Oil with Solid Phase Microextraction and GC–MS

The long-chain polyunsaturated fatty acids (PUFA) found in fish oil, specifically eicosapentanoic acid (EPA) and docosahexanoic acid (DHA) play an important part in human health. As a result, fish oil supplements are commonly consumed by people around the world. Supplements in the form of triacylglycerols (TAG) can be sold at a premium price, compared to those in the ethyl ester (EE) forms. Producers of TAG supplements require a simple, rapid method to determine the authenticity of their raw material. Here, we describe a method to quantify EE in fish oil using solid phase microextraction headspace analysis and GCMS. Despite the variation in linear ranges of the calibration curves with volatility of the EE, 30 individual FA were quantified including common saturated FA such as palmitic and stearic acid, as well as longer chain PUFA, such as EPA and DHA. The method was then applied to three commercial fish oils in the TAG form and two of the products were found to contain EE, with one containing EE above 1.5% w/w, indicating that contamination had occurred. With growing consumer interest in fish oil products, the method proposed here will help resolve future issues of authenticity in fish oils.     

Landfill disposal of CCA-treated wood with construction and demolition (C&D) debris: arsenic, chromium, and copper concentrations in leachate

Although phased out of many residential uses in the United States, the disposal of CCA-treated wood remains a concern because significant quantities have yet to be taken out of service, and it is commonly disposed in landfills. Catastrophic events have also led to the concentrated disposal of CCA-treated wood, often in unlined landfills. The goal of this research was to simulate the complex chemical and biological activity of a construction and demolition (C&D) debris landfill containing a realistic quantity of CCA-treated wood (10% by mass), produce leachate, and then evaluate the arsenic, copper, and chromium concentrations in the leachate as an indication of what may occur in a landfill setting. Copper concentrations were not significantly elevated in the control or experimental simulated landfill setting (alpha = 0.05). However, the concentrations of arsenic and chromium were significantly higher in the experimental simulated landfill leachate compared to the control simulated landfill leachate (alpha = 0.05, p < 0.001). This indicates that disposal of CCA-treated wood with C&D debris can impact leachate quality which, in turn could affect leachate management practices or aquifers below unlined landfills.     

Enhanced Synergism and Mechanism of Action Studies of Synthetic Antimicrobial Metallopeptides

Antimicrobial peptides (AMPs) are found throughout most kingdoms of life, are an important part of host immunity, and have been shown to act synergistically in various organisms to ameliorate bacterial infections. Herein, we report the synergistic behavior observed between two AMPs, Sub5 and CP10A, against E. coli. In addition, enhanced synergistic activity against E. coli and MRSA 43300 for two derivatives of Sub5, extended with the amino-terminal copper and nickel (ATCUN) binding motif, is observed when dosed together with CP10A, while displaying little cytotoxicity towards human dermal fibroblasts. All three combinations of peptides co-localized within bacterial cells as evidenced by fluorescence confocal microscopy. Investigations into the mechanism of synergy shows that all peptides indirectly damage DNA within cells, while only the ATCUN derivatives can oxidize phospholipids. Combinations of peptides were also shown to upregulate the concentration of reactive oxygen species within both E. coli and MRSA 43300. These results suggest that the production of reactive oxygen species is an important aspect mechanistically and further highlights the potential of these metallopeptides to aid in the treatment of antibiotic-resistant infections.     

The impact of imbalanced training data on machine learning for author name disambiguation

Scientometrics - In supervised machine learning for author name disambiguation, negative training data are often dominantly larger than positive training data. This paper examines how the ratios of...     

Leaching of chromated copper arsenate (CCA)-treated wood in a simulated monofill and its potential impacts to landfill leachate

The proper end-of-life management of chromated copper arsenate (CCA)-treated wood, which contains arsenic, copper, and chromium, is a concern to the solid waste management community. Landfills are often the final repository of this waste stream, and the impacts of CCA preservative metals on leachate quality are not well understood. Monofills are a type of landfill designed and operated to dispose a single waste type, such as ash, tires, mining waste, or wood. The feasibility of managing CCA-treated wood in monofills was examined using a simulated landfill (a leaching lysimeter) that contained a mix of new and weathered CCA-treated wood. The liquid to solid ratio (LS) reached in the experiment was 0.63:1. Arsenic, chromium, and copper leached from the lysimeter at average concentrations of 42 mg/L for arsenic, 9.4 mg/L for chromium, and 2.4 mg/L for copper. Complementary batch leaching studies using deionized water were performed on similar CCA-treated wood samples at LS of 5:1 and 10:1. When results from the lysimeter were compared to the batch test results, copper and chromium leachability appeared to be reduced in the lysimeter disposal environment. Of the three metals, arsenic leached to the greatest extent and was found to have the best correlation between the batch and the lysimeter experiments.     

Massively parallel sample preparation for the MALDI MS analyses of tissues

Investigation of the peptidome of the nervous system containing large, often easily identifiable neurons has greatly benefited from single-cell matrix-assisted laser desorption/ionization (MALDI) mass spectrometry and has led to the discovery of hundreds of novel cell-to-cell signaling peptides. By combining new sample preparation methods and established protocols for bioanalytical mass spectrometry, a high-throughput, small-volume approach is created that allows the study of the peptidome of a variety of nervous systems. Specifically, approximately single-cell-sized samples are rapidly prepared from thin tissue slices by adhering the tissue section to a glass bead array that is anchored to a stretchable membrane. Stretching the membrane fragments the tissue slice into thousands of individual samples, their dimensions predominately governed by the size of the individual glass beads. Application of MALDI matrix, followed by the repeated condensation of liquid microdroplets on the fragmented tissue, allows for maximal analyte extraction and incorporation into MALDI matrix crystals. During extraction, analyte migration between the pieces of tissue on separate beads is prevented by the underlying hydrophobic substrate and by controlling the size of the condensation droplets. The procedure, while general in nature, may be tailored to the needs of a variety of analyses, producing mass spectra equivalent to those acquired from single-cell samples.