Identification of genes involved in the mutualistic colonization of the nematode Heterorhabditis bacteriophora by the bacterium Photorhabdus luminescens

<p>Abstract</p> <p>Background</p> <p><it>Photorhabdus </it>are Gram negative entomopathogenic bacteria that also have a mutualistic association with nematodes from the family <it>Heterorhabditis</it>. An essential part of this symbiosis is the ability of the bacterium to colonize the gut of the freeliving form of the nematode called the infective juvenile (IJ). Although the colonization process (also called transmission) has been described phenomonologically very little is known about the underlying molecular mechanisms. Therefore, in this study, we were interested in identifying genes in <it>Photorhabdus </it>that are important for IJ colonization.</p> <p>Results</p> <p>In this work we genetically tagged <it>P. luminescens </it>TT01 with <it>gfp </it>and constructed a library containing over 3200 mutants using the suicide vector, pUT-Km2. Using a combination of <it>in vitro </it>symbiosis assays and fluorescent microscopy we screened this library for mutants that were affected in their ability to colonize the IJ i.e. with decreased transmission frequencies. In total 8 mutants were identified with transmission frequencies of ≤ 30% compared to wild-type. These mutants were mapped to 6 different genetic loci; the <it>pbgPE </it>operon, <it>galE</it>, <it>galU</it>, <it>proQ</it>, <it>asmA </it>and <it>hdfR</it>. The <it>pbgPE</it>, <it>galE </it>and <it>galU </it>mutants were all predicted to be involved in LPS biosynthesis and, in support of this, we have shown that these mutants are avirulent and sensitive to the cationic antimicriobial peptide, polymyxin B. On the other hand the <it>proQ</it>, <it>asmA </it>and <it>hdfR </it>mutants were not affected in virulence and were either as resistant (<it>proQ</it>) or slightly more sensitive (<it>asmA, hdfR</it>) to polymyxin B than the wild-type (WT).</p> <p>Conclusions</p> <p>This is the first report describing the outcome of a comprehensive screen looking for transmission mutants in <it>Photorhabdus</it>. In total 6 genetic loci were identified and we present evidence that all of these loci are involved in the assembly and/or maintenance of LPS and other factors associated with the cell surface. Interestingly several, but not all, of the transmission mutants identified were also avirulent suggesting that there is a significant, but not complete, genetic overlap between pathogenicity and mutualism. Therefore, this study highlights the importance of the cell surface in mediating the symbiotic and pathogenic interactions of <it>Photorhabdus</it>.</p

The role of iron uptake in pathogenicity and symbiosis in Photorhabdus luminescens TT01

<p>Abstract</p> <p>Background</p> <p><it>Photorhabdus </it>are Gram negative bacteria that are pathogenic to insect larvae whilst also having a mutualistic interaction with nematodes from the family <it>Heterorhabditis</it>. Iron is an essential nutrient and bacteria have different mechanisms for obtaining both the ferrous (Fe²⁺) and ferric (Fe³⁺) forms of this metal from their environments. In this study we were interested in analyzing the role of Fe³⁺and Fe²⁺iron uptake systems in the ability of <it>Photorhabdus </it>to interact with its invertebrate hosts.</p> <p>Results</p> <p>We constructed targeted deletion mutants of <it>exbD</it>, <it>feoABC </it>and <it>yfeABCD </it>in <it>P. luminescens </it>TT01. The <it>exbD </it>mutant was predicted to be crippled in its ability to obtain Fe³⁺and we show that this mutant does not grow well in iron-limited media. We also show that this mutant was avirulent to the insect but was unaffected in its symbiotic interaction with <it>Heterorhabditis</it>. Furthermore we show that a mutation in <it>feoABC </it>(encoding a predicted Fe²⁺permease) was unaffected in both virulence and symbiosis whilst the divalent cation transporter encoded by <it>yfeABCD </it>is required for virulence in the Tobacco Hornworm, <it>Manduca sexta </it>(Lepidoptera) but not in the Greater Wax Moth, <it>Galleria mellonella </it>(Lepidoptera). Moreover the Yfe transporter also appears to have a role during colonization of the IJ stage of the nematode.</p> <p>Conclusion</p> <p>In this study we show that iron uptake (via the TonB complex and the Yfe transporter) is important for the virulence of <it>P. luminescens </it>to insect larvae. Moreover this study also reveals that the Yfe transporter appears to be involved in Mn²⁺-uptake during growth in the gut lumen of the IJ nematode. Therefore, the Yfe transporter in <it>P. luminescens </it>TT01 is important during colonization of both the insect and nematode and, moreover, the metal ion transported by this pathway is host-dependent.</p

A Class of Eccentric Binaries with Dynamic Tidal Distortions Discovered with Kepler

We have discovered a class of eccentric binary systems within the Kepler data archive that have dynamic tidal distortions and tidally-induced pulsations. Each has a uniquely shaped light curve that is characterized by periodic brightening or variability at time scales of 4-20 days, frequently accompanied by shorter period oscillations. We can explain the dominant features of the entire class with orbitally-varying tidal forces that occur in close, eccentric binary systems. The large variety of light curve shapes arises from viewing systems at different angles. This hypothesis is supported by spectroscopic radial velocity measurements for five systems, each showing evidence of being in an eccentric binary system. Prior to the discovery of these 17 new systems, only four stars, where KOI-54 is the best example, were known to have evidence of these dynamic tides and tidally-induced oscillations. We perform preliminary fits to the light curves and radial velocity data, present the overall properties of this class and discuss the work required to accurately model these systems.Comment: 13 pages, submitted to Ap

Multi-tissue transcriptomics of the black widow spider reveals expansions, co-options, and functional processes of the silk gland gene toolkit

Background: Spiders (Order Araneae) are essential predators in every terrestrial ecosystem largely because they have evolved potent arsenals of silk and venom. Spider silks are high performance materials made almost entirely of proteins, and thus represent an ideal system for investigating genome level evolution of novel protein functions. However, genomic level resources remain limited for spiders. Results: We de novo assembled a transcriptome for the Western black widow (Latrodectus hesperus) from deeply sequenced cDNAs of three tissue types. Our multi-tissue assembly contained ~100,000 unique transcripts, of which > 27,000 were annotated by homology. Comparing transcript abundance among the different tissues, we identified 647 silk gland-specific transcripts, including the few known silk fiber components (e.g. six spider fibroins, spidroins). Silk gland specific transcripts are enriched compared to the entire transcriptome in several functions, including protein degradation, inhibition of protein degradation, and oxidation-reduction. Phylogenetic analyses of 37 gene families containing silk gland specific transcripts demonstrated novel gene expansions within silk glands, and multiple co-options of silk specific expression from paralogs expressed in other tissues. Conclusions: We propose a transcriptional program for the silk glands that involves regulating gland specific synthesis of silk fiber and glue components followed by protecting and processing these components into functional fibers and glues. Our black widow silk gland gene repertoire provides extensive expansion of resources for biomimetic applications of silk in industry and medicine. Furthermore, our multi-tissue transcriptome facilitates evolutionary analysis of arachnid genomes and adaptive protein systems. Electronic supplementary material The online version of this article (doi:10.1186/1471-2164-15-365) contains supplementary material, which is available to authorized users

“Ne nnipadua mmpe” (the body hates it): Exploring the lived experience of food allergy in Sub-Saharan Africa

The final publication is available at Elsevier via https://dx.doi.org/10.1016/j.socscimed.2018.03.031 © 2018. This manuscript version is made available under the CC-BY-NC-ND 4.0 license https://creativecommons.org/licenses/by-nc-nd/4.0/Allergic diseases have closely followed the rise of non-communicable diseases (NCDs) especially in western societies. As prevalence of NCD is increasing in sub-Saharan Africa (SSA), researchers are hinting that the same future may hold for (food) allergic disease in this world region. Already, researchers are beginning to record prevalence, though with little attention to the social experience of individuals and parents with food allergic children. This paper presents the first qualitative analysis of the daily realities of adults and parents of children with allergies in SSA using Ghana as a case study. Drawing on political ecology of health, this study contextualizes the psychological (e.g. anxiety and fear), social (e.g. stigmatization, social exclusion), and economic (e.g. impact on work & household expenditures) wellbeing of affected persons within the broader sociocultural environment. By exploring the sociocultural environment, the results provide insights into the likely structures (e.g. the lack of familiarity, absence of local discursive repertoire on food allergy, infrastructure deficit) which interact to shape anxiety, and social exclusion of people with allergy. The case study provides evidence suggesting food allergies do have a global reach, and policy makers must heed the message to integrate food allergy into the broader chronic disease prevention agenda

Characterization of electrospun nanocomposite scaffolds and biocompatibility with adipose-derived human mesenchymal stem cells

Electrospun nanocomposite scaffolds were fabricated by encapsulating multi-walled carbon nanotubes (MWNT) in poly (lactic acid) (PLA) nanofibers. Scanning electron microscopy (SEM) confirmed the fabrication of nanofibers, and transmission electron microscopy identified the alignment and dispersion of MWNT along the axis of the fibers. Tensile testing showed an increase in the tensile modulus for a MWNT loading of 0.25 wt% compared with electrospun nanofibrous mats without MWNT reinforcement. Conductivity measurements indicated that the confined geometry of the fibrous system requires only minute doping to obtain significant enhancements at 0.32 wt%. Adipose-derived human mesenchymal stem cells (hMSCs) were seeded on electrospun scaffolds containing 1 wt% MWNT and 0 wt% MWNT, to determine the efficacy of the scaffolds for cell growth, and the effect of MWNT on hMSC viability and proliferation over two weeks in culture. Staining for live and dead cells and DNA quantification indicated that the hMSCs were alive and proliferating through day 14. SEM images of hMSCs at 14 days showed morphological differences, with hMSCs on PLA well spread and hMSCs on PLA with 1% MWNT closely packed and longitudinally aligned

Epidural Analgesia Provides Better Pain Management After Live Liver Donation: A Retrospective Study

Despite the increase in surgical volumes of live liver donation, there has been very little documentation of the postoperative pain experience. The primary aim of this study was to examine the difference in acute postoperative pain intensity and adverse effects between patients who received intravenous patient-controlled analgesia (IV PCA) or patient-controlled epidural analgesia (PCEA) for pain control after live liver donation surgery. A retrospective chart review was performed of 226 consecutive patients who underwent right living donor hepatic surgery at the Toronto General Hospital, Toronto, Canada. Patients who received as their primary postoperative analgesic modality IV PCA (n = 158) were compared to patients who received PCEA (n = 68). Demographic profiles for the 2 groups were similar with respect to age, sex, and body mass index at the time of surgery. For the first 3 postoperative days, pain intensity was significantly lower in patients who received epidural analgesia (P 4) was reported more frequently in the IV PCA group (P < 0.05) along with increased sedation (P < 0.05). Pruritus was reported more frequently in the PCEA group of patients compared to the IV PCA group (P < 0.05). Significant between-group differences were not found for the incidence of postoperative vomiting, the time at which patients began fluid intake, the time to initial ambulation, or the length of hospital stay. In conclusion, epidural analgesia provides better postoperative pain relief, less sedation, but more pruritus than IV PCA after live liver donation

Characterization of an electron conduit between bacteria and the extracellular environment

A number of species of Gram-negative bacteria can use insoluble minerals of Fe(III) and Mn(IV) as extracellular respiratory electron acceptors. In some species of Shewanella, deca-heme electron transfer proteins lie at the extracellular face of the outer membrane (OM), where they can interact with insoluble substrates. To reduce extracellular substrates, these redox proteins must be charged by the inner membrane/periplasmic electron transfer system. Here, we present a spectro-potentiometric characterization of a trans-OM icosa-heme complex, MtrCAB, and demonstrate its capacity to move electrons across a lipid bilayer after incorporation into proteoliposomes. We also show that a stable MtrAB subcomplex can assemble in the absence of MtrC; an MtrBC subcomplex is not assembled in the absence of MtrA; and MtrA is only associated to the membrane in cells when MtrB is present. We propose a model for the modular organization of the MtrCAB complex in which MtrC is an extracellular element that mediates electron transfer to extracellular substrates and MtrB is a trans-OM spanning ß-barrel protein that serves as a sheath, within which MtrA and MtrC exchange electrons. We have identified the MtrAB module in a range of bacterial phyla, suggesting that it is widely used in electron exchange with the extracellular environment