Postembryonic RNAi in Heterorhabditis bacteriophora: a nematode insect parasite and host for insect pathogenic symbionts

Background: Heterorhabditis bacteriophora is applied throughout the world for the biological control of insects and is an animal model to study interspecies interactions, e.g. mutualism, parasitism and vector-borne disease. H. bacteriophora nematodes are mutually associated with the insect pathogen, Photorhabdus luminescens. The developmentally arrested infective juvenile (IJ) stage nematode (vector) specifically transmits Photorhabdus luminescens bacteria (pathogen) in its gut mucosa to the haemocoel of insects (host). The nematode vector and pathogen alone are not known to cause insect disease. RNA interference is an excellent reverse genetic tool to study gene function in C. elegans, and it would be useful in H. bacteriophora to exploit the H. bacteriophora genome project, currently in progress. Results: Soaking L1 stage H. bacteriophora with seven dsRNAs of genes whose C. elegans orthologs had severe RNAi phenotypes resulted in highly penetrant and obvious developmental and reproductive abnormalities. The efficacy of postembryonic double strand RNA interference (RNAi) was evident by abnormal gonad morphology and sterility of adult H. bacteriophora and C. elegans presumable due to defects in germ cell proliferation and gonad development. The penetrance of RNAi phenotypes in H. bacteriophora was high for five genes (87-100%; Hba-cct-2, Hba-daf-21, Hba-icd-1; Hba-nol-5, and Hba-W01G7.3) and moderate for two genes (usually 30-50%; Hba-rack-1 and Hba-arf-1). RNAi of three additional C. elegans orthologs for which RNAi phenotypes were not previously detected in C. elegans, also did not result in any apparent phenotypes in H. bacteriophora. Specific and severe reduction in transcript levels in RNAi treated L1s was determined by quantitative real-time RT-PCR. These results suggest that postembryonic RNAi by soaking is potent and specific. Conclusion: Although RNAi is conserved in animals and plants, RNAi using long dsRNA is not. These results demonstrate that RNAi can be used effectively in H. bacteriophora and can be applied for analyses of nematode genes involved in symbiosis and parasitism. It is likely that RNAi will be an important tool for functional genomics utilizing the high quality draft H. bacteriophora genome sequence

Plasma neuronal specific enolase : a potential stage diagnostic marker in human African trypanosomiasis

© The Author 2014. Published by Oxford University Press on behalf of Royal Society of Tropical Medicine and Hygiene. All rights reserved. For permissions, please e-mail: [email protected]. Funding: This work was supported through grants from the Wellcome Trust [082786] and Foundation for Innovative New Diagnostics.Peer reviewedPublisher PD

Eddington limited starbursts in the central 10pc of AGN, and the Torus in NGC1068

We present results from a survey of nearby AGN using the near infrared adaptive optics integral field spectrograph SINFONI. These data enable us to probe the distribution and kinematics of the gas and stars at spatial resolutions as small as 0.085arcsec. We find strong evidence for recent but short lived starbursts residing in very dense nuclear disks. On scales of less than 10pc these would have reached Eddington-limited luminosities when active, perhaps accounting for their short duration. In addition, for NGC1068 at a resolution of 6pc, we present direct observations of molecular gas close around the AGN which we identify with the obscuring torus.Comment: Conference proceedings to appear in "The Central Engine of Active Galactic Nuclei", ed. L. C. Ho and J.-M. Wang (San Francisco: ASP

Homologous and unique G protein alpha subunits in the nematode Caenorhabditis elegans

A cDNA corresponding to a known G protein alpha subunit, the alpha subunit of Go (Go alpha), was isolated and sequenced. The predicted amino acid sequence of C. elegans Go alpha is 80-87% identical to other Go alpha sequences. An mRNA that hybridizes to the C. elegans Go alpha cDNA can be detected on Northern blots. A C. elegans protein that crossreacts with antibovine Go alpha antibody can be detected on immunoblots. A cosmid clone containing the C. elegans Go alpha gene (goa-1) was isolated and mapped to chromosome I. The genomic fragments of three other C. elegans G protein alpha subunit genes (gpa-1, gpa-2, and gpa-3) have been isolated using the polymerase chain reaction. The corresponding cosmid clones were isolated and mapped to disperse locations on chromosome V. The sequences of two of the genes, gpa-1 and gpa-3, were determined. The predicted amino acid sequences of gpa-1 and gpa-3 are only 48% identical to each other. Therefore, they are likely to have distinct functions. In addition they are not homologous enough to G protein alpha subunits in other organisms to be classified. Thus C. elegans has G proteins that are identifiable homologues of mammalian G proteins as well as G proteins that appear to be unique to C. elegans. Study of identifiable G proteins in C. elegans may result in a further understanding of their function in other organisms, whereas study of the novel G proteins may provide an understanding of unique aspects of nematode physiology

On pseudo-hyperk\"ahler prepotentials

An explicit surjection from a set of (locally defined) unconstrained holomorphic functions on a certain submanifold of (Sp_1(C) \times C^{4n}) onto the set HK_{p,q} of local isometry classes of real analytic pseudo-hyperk\"ahler metrics of signature (4p,4q) in dimension 4n is constructed. The holomorphic functions, called prepotentials, are analogues of K\"ahler potentials for K\"ahler metrics and provide a complete parameterisation of HK_{p,q}. In particular, there exists a bijection between HK_{p,q} and the set of equivalence classes of prepotentials. This affords the explicit construction of pseudo-hyperk\"ahler metrics from specified prepotentials. The construction generalises one due to Galperin, Ivanov, Ogievetsky and Sokatchev. Their work is given a coordinate-free formulation and complete, self-contained proofs are provided. An appendix provides a vital tool for this construction: a reformulation of real analytic G-structures in terms of holomorphic frame fields on complex manifolds.Comment: 53 pages; v2: minor amendments to Def.4.1 and Theorem 4.5; a paragraph inserted in the proof of the latter; V3: minor changes; V4: minor changes/ typos corrected for journal versio

Mid-Infrared line diagnostics of Active Galaxies -- A spectroscopic AGN survey with ISO-SWS

We present medium resolution (R approx. 1500) ISO-SWS 2.4--45 micron spectra of a sample of 29 galaxies with active nuclei. This data set is rich in fine structure emission lines tracing the narrow line regions and (circum-)nuclear star formation regions, and it provides a coherent spectroscopic reference for future extragalactic studies in the mid-infrared. We use the data set to briefly discuss the physical conditions in the narrow line regions (density, temperature, excitation, line profiles) and to test for possible differences between AGN sub-types. Our main focus is on new tools for determining the propertibes of dusty galaxies and on the AGN-starburst connection. We present mid-IR line ratio diagrams which can be used to identify composite (starburst + AGN) sources and to distinguish between emission excited by active nuclei and emission from (circum-nuclear) star forming regions. For instance, line ratios of high to low excitation lines like [O IV]25.9um/[Ne II]12.8um, that have been used to probe for AGNs in dusty objects, can be examined in more detail and with better statistics now. In addition, we present two-dimensional diagnostic diagrams that are fully analogous to classical optical diagnostic diagrams, but better suited for objects with high extinction. Finally, we discuss correlations of mid-infrared line fluxes to the mid- and far-infrared continuum. We compare these relations to similar relations in starburst galaxies in order to examine the contribution of AGNs to the bolometric luminosities of their host galaxies. The spectra are available in electronic form from the authors.Comment: 24 pages, 23 figures, 5 tables. Accepted for A&