The Lymantria dispar IPLB-Ld652Y Cell Line Transcriptome Comprises Diverse Virus-Associated Transcripts

The enhanced viral susceptibility of the gypsy moth (Lymantria dispar)-derived IPLB-Ld652Y cell line has made it a popular in vitro system for studying virus-related phenomena in the Lepidoptera. Using both single-pass EST sequencing and 454-based pyrosequencing, a transcriptomic library of 14,368 putatively unique transcripts (PUTs) was produced comprising 8,476,050 high-quality, informative bases. The gene content of the IPLB-Ld652Y transcriptome was broadly assessed via comparison with the NCBI non-redundant protein database, and more detailed functional annotation was inferred by comparison to the Swiss-Prot subset of UniProtKB. In addition to L. dispar cellular transcripts, a diverse array of both RNA and DNA virus-associated transcripts was identified within the dataset, suggestive of a high level of viral expression and activity in IPLB-Ld652Y cells. These sequence resources will provide a sound basis for developing testable experimental hypotheses by insect virologists, and suggest a number of avenues for potential research

Identification of the water quality factors which prevent fingernail clams from recolonizing the Illinois River—phase III

TECHNICAL COMPLETION REPORT Project No. B-124-ILL Agreement No. 14-34-0001-0217The purpose of this research was to determine why fingernail clams have been unable to recolonize a 100-mile reach of the Illinois River where they were abundant prior to a die-off in the 1950's. Fingernail clams are major links in food chains leading from detritus and algae to higher level consumers valued by man, such as fish and water fowl. Three suspected toxicants and sediments from the reach where the die-off occurred were tested on intact fingernail clams (Musculium transversum) and gill preparations isolated from the clams. Concentrations of fluoride, lead and cadmium which caused a 50% reduction in the rate of beating of cilia on isolated clam gills, after 10 minutes of exposure (10-minute EC50), were 0.75, 0.02 and 0.06 mg/l , respectively. Mixtures of cadmium and fluoride were slightly more toxic to clam gills than predicted from results of bioassays with single toxicants. A fluoride concentration of 2.82 mg/l killed intact fingernail clams after eight weeks of exposure, while mortality in lesser concentrations and one higher concentration did not differ significantly from controls maintained in well water to which no fluoride had been added. Hence, the sub-lethal response exhibited by the gills is at least four times more sensitive than the lethal response. Maximum fluoride concentrations reported by the U.S. Geological Survey at two stations in the Illinois River ranged from .6 to .8 mg/l between 1979 and 1981, considerably below the concentrations which affected growth and survival of intact clams during 8-week exposures in our laboratory, but slightly above the level which affected isolated clam gills. A lead bioassay using intact clams was completed, but the results were ambiguous because concentration ranges in separate test chambers overlapped. In addition, insoluble lead precipitates accumulated in the test chambers, and the relative toxicity to clams of the soluble versus the insoluble lead was not determined. Until additional bioassays are completed, it is impossible to determine whether the maximum total lead concentrations of 0.40 mg/l which occurred in the Illinois River between 1979 and 1981 could have contributed to the failure of fingernail clams to recolonize the river. Fingernail clams exposed to sediments from lakes along the Illinois River suffered greater mortality after six weeks of exposure than clams exposed to sediment from the Mississippi River, although the differences were not statistically significant. The same sediments tested on clam gills produced statistically significant changes in ciliary beating rate and particle transport rates on the gills. Sediments from the upstream lakes cause a greater depression in the particle transport rate and ciliary beating rates than sediments from downstream lakes. In addition, sediments from the lake furthest upstream caused a drastic change from the normal metachronal beating pattern to an atypical synchronous pattern. The results with the gill assay suggest that sediments in the Illinois River contain unidentified toxic factors and that sediments in the upper river, closer to the metropolitan areas of Joliet and Chicago, are more toxic than sediments further downstream. These results should be confirmed by additional tests with intact clams, including field tests with caged organisms. Parallel chemical analyses and bioassays of extracts from the sediments should be performed to identify the toxic components.U.S. Department of the InteriorU.S. Geological SurveyOpe

De novo formation of an aggregation pheromone precursor by an isoprenyl diphosphate synthase-related terpene synthase in the harlequin bug.

Insects use a diverse array of specialized terpene metabolites as pheromones in intraspecific interactions. In contrast to plants and microbes, which employ enzymes called terpene synthases (TPSs) to synthesize terpene metabolites, limited information from few species is available about the enzymatic mechanisms underlying terpene pheromone biosynthesis in insects. Several stink bugs (Hemiptera: Pentatomidae), among them severe agricultural pests, release 15-carbon sesquiterpenes with a bisabolene skeleton as sex or aggregation pheromones. The harlequin bug, Murgantia histrionica, a specialist pest of crucifers, uses two stereoisomers of 10,11-epoxy-1-bisabolen-3-ol as a male-released aggregation pheromone called murgantiol. We show that MhTPS (MhIDS-1), an enzyme unrelated to plant and microbial TPSs but with similarity to trans-isoprenyl diphosphate synthases (IDS) of the core terpene biosynthetic pathway, catalyzes the formation of (1S,6S,7R)-1,10-bisaboladien-1-ol (sesquipiperitol) as a terpene intermediate in murgantiol biosynthesis. Sesquipiperitol, a so-far-unknown compound in animals, also occurs in plants, indicating convergent evolution in the biosynthesis of this sesquiterpene. RNAi-mediated knockdown of MhTPS mRNA confirmed the role of MhTPS in murgantiol biosynthesis. MhTPS expression is highly specific to tissues lining the cuticle of the abdominal sternites of mature males. Phylogenetic analysis suggests that MhTPS is derived from a trans-IDS progenitor and diverged from bona fide trans-IDS proteins including MhIDS-2, which functions as an (E,E)-farnesyl diphosphate (FPP) synthase. Structure-guided mutagenesis revealed several residues critical to MhTPS and MhFPPS activity. The emergence of an IDS-like protein with TPS activity in M. histrionica demonstrates that de novo terpene biosynthesis evolved in the Hemiptera in an adaptation for intraspecific communication

Maternal Blood Lipid Biomarkers of Oligodendrocyte Pathology to Predict Fetal Alcohol Spectrum Disorders

INTRODUCTION: Up to 9.9% of children have fetal alcohol spectrum disorders (FASD), the most frequent cause of intellectual disability in the US. FASD may involve abnormal brain development, including dysmyelination, suggesting abnormal development of oligodendrocytes (OLs), which make myelin and are rich in lipids. Indeed, low serum levels of omega-3 fatty acids (ω-3) have been reported in FASD. Free fatty acids bind to specific receptors (FFARs). We have isolated cell type-specific fetal brain-derived exosomes (FB-E) from maternal blood and sampled their contents to search for lipid-related biomarkers that predict FASD. METHODS: Blood samples were collected from two groups of pregnant women: 1) those who consumed EtOH during pregnancy, and 2) non-EtOH using controls, under an IRB-approved protocol. Serum and OL-derived exosomes (OL-Es) were used to assay myelin basic protein (MBP) and FFAR by ELISA and droplet digital PCR (ddPCR), respectively. RESULTS: FFAR and MBP proteins were downregulated in the EtOH group compared to controls, and this difference was greatest in OL-Es from maternal blood compared maternal serum. CONCLUSION: MBP and FFAR levels were reduced in OL-Es from EtOH-consuming pregnant women. The data suggest potential therapeutic targets to predict which children are at risk for developing FASD and reduce dysmyelination in developing

Social media for research discourse, dissemination, and collaboration in rheumatology

Social media has become an important venue for rheumatologists, patients, organizations, and other stakeholders to discuss recent research advances in diagnosis and management of rheumatic disorders. In this article, we describe the current state of how social media may enhance dissemination, discourse, and collaboration in rheumatology research. Social media may refer to social platforms like Twitter and Instagram or digital media like podcasts and other websites that are operated for providing as free, open-access medical education (FOAM). Twitter has been one of the most active social media venues and continues to host a vibrant rheumatology community. Examples of research discussions on Twitter include organic user tweets, educational threads ( tweetorials ), live-tweeting academic conferences, and journals posting recently-accepted articles. Some research collaborations have been initiated through social media interactions. Social media may also directly contribute to research by facilitating the recruitment of study participants and the collection of survey-based data. Thus, social media is an evolving and important tool to enhance research discourse, dissemination, and collaboration in rheumatology

Heterogeneous N2O5 Uptake During Winter: Aircraft Measurements During the 2015 WINTER Campaign and Critical Evaluation of Current Parameterizations

Nocturnal dinitrogen pentoxide (N2O5) heterogeneous chemistry impacts regional air quality and the distribution and lifetime of tropospheric oxidants. Formed from the oxidation of nitrogen oxides, N2O5 is heterogeneously lost to aerosol with a highly variable reaction probability, γ(N2O5), dependent on aerosol composition and ambient conditions. Reaction products include soluble nitrate (HNO3 or NO3−) and nitryl chloride (ClNO2). We report the first‐ever derivations of γ(N2O5) from ambient wintertime aircraft measurements in the critically important nocturnal residual boundary layer. Box modeling of the 2015 Wintertime INvestigation of Transport, Emissions, and Reactivity (WINTER) campaign over the eastern United States derived 2,876 individual γ(N2O5) values with a median value of 0.0143 and range of 2 × 10−5 to 0.1751. WINTER γ(N2O5) values exhibited the strongest correlation with aerosol water content, but weak correlations with other variables, such as aerosol nitrate and organics, suggesting a complex, nonlinear dependence on multiple factors, or an additional dependence on a nonobserved factor. This factor may be related to aerosol phase, morphology (i.e., core shell), or mixing state, none of which are commonly measured during aircraft field studies. Despite general agreement with previous laboratory observations, comparison of WINTER data with 14 literature parameterizations (used to predict γ(N2O5) in chemical transport models) confirms that none of the current methods reproduce the full range of γ(N2O5) values. Nine reproduce the WINTER median within a factor of 2. Presented here is the first field‐based, empirical parameterization of γ(N2O5), fit to WINTER data, based on the functional form of previous parameterizations

V838 Monocerotis: A Geometric Distance from Hubble Space Telescope Polarimetric Imaging of its Light Echo

Following the outburst of the unusual variable star V838 Monocerotis in 2002, a spectacular light echo appeared. A light echo provides the possibility of direct geometric distance determination, because it should contain a ring of highly linearly polarized light at a linear radius of ct, where t is the time since the outburst. We present imaging polarimetry of the V838 Mon light echo, obtained in 2002 and 2005 with the Advanced Camera for Surveys onboard the Hubble Space Telescope, which confirms the presence of the highly polarized ring. Based on detailed modeling that takes into account the outburst light curve, the paraboloidal echo geometry, and the physics of dust scattering and polarization, we find a distance of 6.1+-0.6 kpc. The error is dominated by the systematic uncertainty in the scattering angle of maximum linear polarization, taken to be theta_{max}=90^o +- 5^o. The polarimetric distance agrees remarkably well with a distance of 6.2+-1.5 kpc obtained from the entirely independent method of main-sequence fitting to a sparse star cluster associated with V838 Mon. At this distance, V838 Mon at maximum light had M_V\simeq-9.8, making it temporarily one of the most luminous stars in the Local Group. Our validation of the polarimetric method offers promise for measurement of extragalactic distances using supernova light echoes.Comment: 43 pages, 17 figures, 3 tables; accepted for publication in the Astronomical Journal. Version with high-quality figures available at http://www.stsci.edu/~bond/v838monpolariz.pd

Exosomal Lipid Biomarkers of Oligodendrocyte Pathology to Predict Scoliosis in Children with Cerebral Palsy

INTRODUCTION: Cerebral Palsy (CP), the most common cause of disability in children, is phenotypically heterogeneous. Approximately 20% of cases develop severe scoliosis. A pathological hallmark of CP is periventricular leukomalacia (PVL), which is due to dysmyelination, suggesting the possibility of a lipidomic abnormality. Risk factors for CP include perinatal hypoxia, prematurity, multiple gestation, ischemia, infection, and maternal alcohol consumption. There is evidence for low serum levels of omega-3 (ω-3) fatty acids in CP patients, and separately in idiopathic scoliosis. Many effects of free fatty acids (FFAs) are mediated via specific G protein-coupled free fatty acid receptors (FFARs), which play essential roles as nutritional and signaling molecules. FFAs, including ω-3, and their receptors are involved in the development and metabolism of oligodendrocytes (OLs), and are critical to myelination. Thus, the cases of CP that will develop severe scoliosis might be those in which there is a deficiency of ω-3, FFARs, or other lipidomic abnormality that is detectable early in the plasma. If so, we might be able to predict scoliosis and prevent it with dietary supplementation. METHODS: Blood samples were collected from four groups of patients at the Philadelphia Shriners Children\u27s Hospital (SCH-P): 1) patients with CP; 2) severe scoliosis (\u3e40o); 3) CP plus scoliosis; and 4) non-impaired controls stratified by age (2-18 yrs), gender, and race/ethnicity, under an IRB-approved protocol. Serum proteins and RNA were purified, and OL-derived exosomes (OL-Es) isolated, using myelin basic protein (MBP) as a late OL marker. Protein was used for the detection of MBP and FFAR by enzyme-linked immunosorbent assays (ELISAs), and by flow cytometry. RNA was assayed by digital droplet polymerase chain reaction (ddPCR) for OL markers and FFAR expression. RESULTS: FFAR and MBP proteins were downregulated in each of the three patient groups compared to controls, and this difference was greatest in both patients with CP plus scoliosis. CONCLUSION: Altogether, MBP and FFAR levels were reduced in OL-Es from both children with CP plus scoliosis. The lipid abnormalities specific to CP with scoliosis were concentrated in OLs. Our data might i) suggest therapeutic targets to reduce dysmyelination and scoliosis in CP, ii) predict which children are at risk for developing scoliosis, iii) lead to therapeutic trials of fatty acids for CP and other dysmyelinating neurological disorders

The first crop plant genetically engineered to release an insect pheromone for defence

Insect pheromones offer potential for managing pests of crop plants. Volatility and instability are problems for deployment in agriculture but could be solved by expressing genes for the biosynthesis of pheromones in the crop plants. This has now been achieved by genetically engineering a hexaploid variety of wheat to release (E)-β-farnesene (Eβf), the alarm pheromone for many pest aphids, using a synthetic gene based on a sequence from peppermint with a plastid targeting amino acid sequence, with or without a gene for biosynthesis of the precursor farnesyl diphosphate. Pure Eβf was produced in stably transformed wheat lines with no other detectable phenotype but requiring targeting of the gene produced to the plastid. In laboratory behavioural assays, three species of cereal aphids were repelled and foraging was increased for a parasitic natural enemy. Although these studies show considerable potential for aphid control, field trials employing the single and double constructs showed no reduction in aphids or increase in parasitism. Insect numbers were low and climatic conditions erratic suggesting the need for further trials or a closer imitation, in the plant, of alarm pheromone release

A single vertebrate DNA virus protein disarms invertebrate immunity to RNA virus infection

Virus-host interactions drive a remarkable diversity of immune responses and countermeasures. We found that two RNA viruses with broad host ranges, vesicular stomatitis virus (VSV) and Sindbis virus (SINV), are completely restricted in their replication after entry into Lepidopteran cells. This restriction is overcome when cells are co-infected with vaccinia virus (VACV), a vertebrate DNA virus. Using RNAi screening, we show that Lepidopteran RNAi, Nuclear Factor-kappaB, and ubiquitin-proteasome pathways restrict RNA virus infection. Surprisingly, a highly conserved, uncharacterized VACV protein, A51R, can partially overcome this virus restriction. We show that A51R is also critical for VACV replication in vertebrate cells and for pathogenesis in mice. Interestingly, A51R colocalizes with, and stabilizes, host microtubules and also associates with ubiquitin. We show that A51R promotes viral protein stability, possibly by preventing ubiquitin-dependent targeting of viral proteins for destruction. Importantly, our studies reveal exciting new opportunities to study virus-host interactions in experimentally-tractable Lepidopteran systems