Science.gov

Sample records for castaneum em milho

  1. Nuclear structures in Tribolium castaneum oocytes.

    PubMed

    Bogolyubov, Dmitry S; Batalova, Florina M; Kiselyov, Artyom M; Stepanova, Irina S

    2013-10-01

    The first ultrastructural and immunomorphological characteristics of the karyosphere (karyosome) and extrachromosomal nuclear bodies in the red flour beetle, Tribolium castaneum, are presented. The karyosphere forms early in the diplotene stage of meiotic prophase by the gathering of all oocyte chromosomes in a limited nuclear volume. Using the BrUTP assay, T. castaneum oocyte chromosomes united in the karyosphere maintain their transcriptional activity until the end of oocyte growth. Hyperphosphorylated RNA polymerase II and basal transcription factors (TFIID and TFIIH) were detected in the perichromatin region of the karyosphere. The T. castaneum karyosphere has an extrachromosomal capsule that separates chromosomes from the rest of the nucleoplasm. Certain structural proteins (F-actin, lamin B) were found in the capsule. Unexpectedly, the karyosphere capsule in T. castaneum oocytes was found to be enriched in TMG-capped snRNAs, which suggests that the capsule is not only a structural support for the karyosphere, but may be involved in biogenesis of snRNPs. We also identified the counterparts of 'universal' extrachromosomal nuclear domains, Cajal bodies (CBs) and interchromatin granule clusters (IGCs). Nuclear bodies containing IGC marker protein SC35 display some features unusual for typical IGCs. SC35 domains in T. castaneum oocytes are predominantly fibrillar complex bodies that do not contain trimethyl guanosine (TMG)-capped small nuclear (sn) RNAs. Microinjections of 2'-O-methyl (U)22 probes into the oocytes allowed revealing poly(A)+ RNAs in these nuclear domains. Several proteins related to mRNA export (heterogeneous ribonucleoprotein core protein A1, export adapters Y14 and Aly and export receptor NXF1) were also detected there. We believe that unusual SC35 nuclear domains of T. castaneum oocytes are possibly involved in mRNP but not snRNP biogenesis.

  2. [Karyosphere capsule in Tribolium castaneum oocytes].

    PubMed

    Batalova, F M; Bogoliubov, D S

    2013-01-01

    Structure and composition of the karyosphere (karyosome) capsule were studied in the oocytes of a laboratory insect, Tribolium castaneum, with the use of electron microscopy and immunoelectron cytochemistry. Basing on the study of nuclear structure dynamics, we distinguished 8 stages that characterize the period of oocyte growth. At the diplotene stage, T. castaneum oocyte chromosomes conjoin early into a compact karyosphere, but a significant chromatin condensation does not occur. The process of karyosphere formation is accompanied by the development of an extensive extrachromosome capsule surrounding chromatin. The capsule consists of a material of different morphological types. Significant molecular components of the T. castaneum karyosphere capsule are represented by the proteins of nuclear matrix including F-actin and lamin B. Besides the structural proteins, the Sm proteins of small nuclear (sn) RNPs and mature 2,2,7-trimethyl guanosine (TMG) 5'-capped snRNAs are revealed immunocytochemically in the karyosphere capsule. The obtained data can form a basis for further expansion of ideas on the functions of the karyosphere capsule as a specialized extrachromosomal nuclear domain of the oocytes. We believe that the T. castaneum karyosphere capsule plays not only a structural role, but may be involved directly in the processes related to gene expression.

  3. Genetic structure of Tribolium castaneum (Coleptera: Tenebrionidae) populations in mills

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The red flour beetle, Tribolium castaneum, is primarily found associated with human structures such as wheat and rice mills, which are spatially isolated resource patches with apparently limited immigration that could produce genetically structured populations. We investigated genetic diversity and...

  4. Larval RNA Interference in the Red Flour Beetle, Tribolium castaneum

    PubMed Central

    Tomoyasu, Yoshinori

    2014-01-01

    The red flour beetle, Tribolium castaneum, offers a repertoire of experimental tools for genetic and developmental studies, including a fully annotated genome sequence, transposon-based transgenesis, and effective RNA interference (RNAi). Among these advantages, RNAi-based gene knockdown techniques are at the core of Tribolium research. T. castaneum show a robust systemic RNAi response, making it possible to perform RNAi at any life stage by simply injecting double-stranded RNA (dsRNA) into the beetle’s body cavity. In this report, we provide an overview of our larval RNAi technique in T. castaneum. The protocol includes (i) isolation of the proper stage of T. castaneum larvae for injection, (ii) preparation for the injection setting, and (iii) dsRNA injection. Larval RNAi is a simple, but powerful technique that provides us with quick access to loss-of-function phenotypes, including multiple gene knockdown phenotypes as well as a series of hypomorphic phenotypes. Since virtually all T. castaneum tissues are susceptible to extracellular dsRNA, the larval RNAi technique allows researchers to study a wide variety of tissues in diverse contexts, including the genetic basis of organismal responses to the outside environment. In addition, the simplicity of this technique stimulates more student involvement in research, making T. castaneum an ideal genetic system for use in a classroom setting. PMID:25350485

  5. Larval RNA interference in the red flour beetle, Tribolium castaneum.

    PubMed

    Linz, David M; Clark-Hachtel, Courtney M; Borràs-Castells, Ferran; Tomoyasu, Yoshinori

    2014-10-13

    The red flour beetle, Tribolium castaneum, offers a repertoire of experimental tools for genetic and developmental studies, including a fully annotated genome sequence, transposon-based transgenesis, and effective RNA interference (RNAi). Among these advantages, RNAi-based gene knockdown techniques are at the core of Tribolium research. T. castaneum show a robust systemic RNAi response, making it possible to perform RNAi at any life stage by simply injecting double-stranded RNA (dsRNA) into the beetle's body cavity. In this report, we provide an overview of our larval RNAi technique in T. castaneum. The protocol includes (i) isolation of the proper stage of T. castaneum larvae for injection, (ii) preparation for the injection setting, and (iii) dsRNA injection. Larval RNAi is a simple, but powerful technique that provides us with quick access to loss-of-function phenotypes, including multiple gene knockdown phenotypes as well as a series of hypomorphic phenotypes. Since virtually all T. castaneum tissues are susceptible to extracellular dsRNA, the larval RNAi technique allows researchers to study a wide variety of tissues in diverse contexts, including the genetic basis of organismal responses to the outside environment. In addition, the simplicity of this technique stimulates more student involvement in research, making T. castaneum an ideal genetic system for use in a classroom setting.

  6. Anatomical localization and stereoisomeric composition of Tribolium castaneum aggregation pheromones

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We report that the abdomen and associated tissues are the predominant sources of male-produced pheromones in the red flour beetle, Tribolium castaneum, and for the first time describe the stereoisomeric composition of the natural blend of isomers of the aggregation pheromone 4,8-dimethyldecanal (DMD...

  7. Movement of Tribolium castaneum within a flour mill

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The colonization of food processing plants by stored-product pests and their distribution within a facility depend, in part, on their dispersal ability. In this case study, we relied on self-mark recapture to evaluate the ability of Tribolium castaneum, the red flour beetle, to move among floors wit...

  8. UVB Radiation Delays Tribolium castaneum Metamorphosis by Influencing Ecdysteroid Metabolism

    PubMed Central

    Sang, Wen; Yu, Lin; He, Li; Ma, Wei-Hua; Zhu, Zhi-Hui; Zhu, Fen; Wang, Xiao-Ping; Lei, Chao-Liang

    2016-01-01

    Ultraviolet B (UVB) radiation is an important environmental factor. It is generally known that UVB exhibits high genotoxicity due to causing DNA damage, potentially leading to skin carcinogenesis and aging in mammals. However, little is known about the effects of UVB on the development and metamorphosis of insects, which are the most abundant terrestrial animals. In the present study, we performed dose-response analyses of the effects UVB irradiation on Tribolium castaneum metamorphosis, assessed the function of the T. castaneum prothoracicotropic hormone gene (Trcptth), and analyzed ecdysteroid pathway gene expression profile and ecdysterone titers post-UVB irradiation. The results showed that UVB not only caused death of T. castaneum larvae, but also delayed larval-pupal metamorphosis and reduced the size and emergence rate of pupae. In addition, we verified the function of Trcptth, which is responsible for regulating metamorphosis. It was also found that the expression profiles of Trcptth as well as ecdysteroidogenesis and response genes were influenced by UVB radiation. Therefore, a disturbance pulse of ecdysteroid may be involved in delaying development under exposure to irradiation. To our knowledge, this is the first report indicating that UVB can influence the metamorphosis of insects. This study will contribute to a better understanding of the impact of UVB on signaling mechanisms in insect metamorphosis. PMID:26986217

  9. UVB Radiation Delays Tribolium castaneum Metamorphosis by Influencing Ecdysteroid Metabolism.

    PubMed

    Sang, Wen; Yu, Lin; He, Li; Ma, Wei-Hua; Zhu, Zhi-Hui; Zhu, Fen; Wang, Xiao-Ping; Lei, Chao-Liang

    2016-01-01

    Ultraviolet B (UVB) radiation is an important environmental factor. It is generally known that UVB exhibits high genotoxicity due to causing DNA damage, potentially leading to skin carcinogenesis and aging in mammals. However, little is known about the effects of UVB on the development and metamorphosis of insects, which are the most abundant terrestrial animals. In the present study, we performed dose-response analyses of the effects UVB irradiation on Tribolium castaneum metamorphosis, assessed the function of the T. castaneum prothoracicotropic hormone gene (Trcptth), and analyzed ecdysteroid pathway gene expression profile and ecdysterone titers post-UVB irradiation. The results showed that UVB not only caused death of T. castaneum larvae, but also delayed larval-pupal metamorphosis and reduced the size and emergence rate of pupae. In addition, we verified the function of Trcptth, which is responsible for regulating metamorphosis. It was also found that the expression profiles of Trcptth as well as ecdysteroidogenesis and response genes were influenced by UVB radiation. Therefore, a disturbance pulse of ecdysteroid may be involved in delaying development under exposure to irradiation. To our knowledge, this is the first report indicating that UVB can influence the metamorphosis of insects. This study will contribute to a better understanding of the impact of UVB on signaling mechanisms in insect metamorphosis.

  10. UVB Radiation Delays Tribolium castaneum Metamorphosis by Influencing Ecdysteroid Metabolism.

    PubMed

    Sang, Wen; Yu, Lin; He, Li; Ma, Wei-Hua; Zhu, Zhi-Hui; Zhu, Fen; Wang, Xiao-Ping; Lei, Chao-Liang

    2016-01-01

    Ultraviolet B (UVB) radiation is an important environmental factor. It is generally known that UVB exhibits high genotoxicity due to causing DNA damage, potentially leading to skin carcinogenesis and aging in mammals. However, little is known about the effects of UVB on the development and metamorphosis of insects, which are the most abundant terrestrial animals. In the present study, we performed dose-response analyses of the effects UVB irradiation on Tribolium castaneum metamorphosis, assessed the function of the T. castaneum prothoracicotropic hormone gene (Trcptth), and analyzed ecdysteroid pathway gene expression profile and ecdysterone titers post-UVB irradiation. The results showed that UVB not only caused death of T. castaneum larvae, but also delayed larval-pupal metamorphosis and reduced the size and emergence rate of pupae. In addition, we verified the function of Trcptth, which is responsible for regulating metamorphosis. It was also found that the expression profiles of Trcptth as well as ecdysteroidogenesis and response genes were influenced by UVB radiation. Therefore, a disturbance pulse of ecdysteroid may be involved in delaying development under exposure to irradiation. To our knowledge, this is the first report indicating that UVB can influence the metamorphosis of insects. This study will contribute to a better understanding of the impact of UVB on signaling mechanisms in insect metamorphosis. PMID:26986217

  11. A cell line derived from the red flour beetle Tribolium castaneum (Coleoptera: Tenebrionidae)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The red flour beetle, Tribolium castaneum, is a model organism for agricultural and medical research and its complete genome is sequenced. We established a continuously replicating T. castaneum cell line to complement existing physiological, genetic and genomic research tools. We set up trial cell ...

  12. A Family of Chemoreceptors in Tribolium castaneum (Tenebrionidae: Coleoptera)

    PubMed Central

    Abdel-latief, Mohatmed

    2007-01-01

    Chemoperception in invertebrates is mediated by a family of G-protein-coupled receptors (GPCR). To date nothing is known about the molecular mechanisms of chemoperception in coleopteran species. Recently the genome of Tribolium castaneum was sequenced for use as a model species for the Coleoptera. Using blast searches analyses of the T. castaneum genome with previously predicted amino acid sequences of insect chemoreceptor genes, a putative chemoreceptor family consisting of 62 gustatory receptors (Grs) and 26 olfactory receptors (Ors) was identified. The receptors have seven transmembrane domains (7TMs) and all belong to the GPCR receptor family. The expression of the T. castaneum chemoreceptor genes was investigated using quantification real- time RT-PCR and in situ whole mount RT-PCR analysis in the antennae, mouth parts, and prolegs of the adults and larvae. All of the predicted TcasGrs were expressed in the labium, maxillae, and prolegs of the adults but TcasGr13, 19, 28, 47, 62, 98, and 61 were not expressed in the prolegs. The TcasOrs were localized only in the antennae and not in any of the beetles gustatory organs with one exception; the TcasOr16 (like DmelOr83b), which was localized in the antennae, labium, and prolegs of the beetles. A group of six TcasGrs that presents a lineage with the sugar receptors subfamily in Drosophila melanogaster were localized in the lacinia of the Tribolium larvae. TcasGr1, 3, and 39, presented an ortholog to CO2 receptors in D. melanogaster and Anopheles gambiae was recorded. Low expression of almost all of the predicted chemoreceptor genes was observed in the head tissues that contain the brains and suboesophageal ganglion (SOG). These findings demonstrate the identification of a chemoreceptor family in Tribolium, which is evolutionarily related to other insect species. PMID:18091992

  13. Anatomical localization and stereoisomeric composition of Tribolium castaneum aggregation pheromones

    NASA Astrophysics Data System (ADS)

    Lu, Yujie; Beeman, Richard W.; Campbell, James F.; Park, Yoonseong; Aikins, Michael J.; Mori, Kenji; Akasaka, Kazuaki; Tamogami, Shigeyuki; Phillips, Thomas W.

    2011-09-01

    We report that the abdominal epidermis and associated tissues are the predominant sources of male-produced pheromones in the red flour beetle, Tribolium castaneum and, for the first time, describe the stereoisomeric composition of the natural blend of isomers of the aggregation pheromone 4,8-dimethyldecanal (DMD) in this important pest species. Quantitative analyses via gas chromatography-mass spectrometry showed that the average amount of DMD released daily by single feeding males of T. castaneum was 878 ± 72 ng (SE). Analysis of different body parts identified the abdominal epidermis as the major source of aggregation pheromone; the thorax was a minor source, while no DMD was detectable in the head. No internal organs or obvious male-specific glands were associated with pheromone deposition. Complete separation of all four stereoisomers of DMD was achieved following oxidation to the corresponding acid, derivatization with (1 R, 2 R)- and (1 S, 2 S)-2-(anthracene-2,3-dicarboximido)cyclohexanol to diastereomeric esters, and their separation on reversed-phase high-performance liquid chromatography at -54°C. Analysis of the hexane eluate from Porapak-Q-collected volatiles from feeding males revealed the presence of all four isomers (4 R,8 R)/(4 R,8 S)/(4 S,8 R)/(4 S,8 S) at a ratio of approximately 4:4:1:1. A walking orientation bioassay in a wind tunnel with various blends of the four synthetic isomers further indicated that the attractive potency of the reconstituted natural blend of 4:4:1:1 was equivalent to that of the natural pheromone and greater than that of the 1:1 blend of (4 R,8 R)/(4 R,8 S) used in commercial lures.

  14. Tribolium castaneum larval gut transcriptome and proteome: A resource for the study of the coleopteran gut.

    PubMed

    Morris, Kaley; Lorenzen, Marcé D; Hiromasa, Yasuaki; Tomich, John M; Oppert, Cris; Elpidina, Elena N; Vinokurov, Konstantin; Jurat-Fuentes, Juan Luis; Fabrick, Jeff; Oppert, Brenda

    2009-08-01

    Tribolium castaneum is an important agricultural pest and an advanced genetic model for coleopteran insects. We have taken advantage of the recently acquired T. castaneum genome to identify T. castaneum genes and proteins in one of the more critical environmental interfaces of the insect, the larval alimentary tract. Genetic transcripts isolated from the T. castaneum larval gut were labeled and hybridized to a custom array containing oligonucleotides from predicted genes in the T. castaneum genome. Through a ranking procedure based on relative labeling intensity, we found that approximately 17.6% of the genes represented in the array were predicted to be highly expressed in gut tissue. Several genes were selected to compare relative expression levels in larval gut, head, or carcass tissues using quantitative real-time PCR, and expression levels were, with few exceptions, consistent with the gut rankings. In parallel with the microarrays, proteins extracted from the T. castaneum larval gut were subjected to proteomic analysis. Two-dimensional electrophoretic analysis combined with MALDI-TOF resulted in the identification of 37 of 88 selected protein samples. As an alternative strategy, one-dimensional electrophoretic separation of T. castaneum larval gut proteins followed by two-dimensional nano-HPLC and ESI-MS/MS resulted in the identification of 98 proteins. A comparison of the proteomic studies indicated that 16 proteins were commonly identified in both, whereas 80 proteins from the proteomic analyses corresponded to genes with gut rankings indicative of high expression in the microarray analysis. These data serve as a resource of T. castaneum transcripts and proteins in the larval gut and provide the basis for comparative transcriptomic and proteomic studies related to the gut of coleopteran insects.

  15. Effects of microwave radiation and conductive heating on Tribolium castaneum microstructure.

    PubMed

    Lu, H H; Zhou, J C; Yan, D; Zhao, S M; Xiong, S B

    2011-01-01

    Microwave radiation and conductive heating were used to completely kill adult Tribolium castaneum (Coleoptera: Tenebrionidae) in wheat flour to protect the flour during storage without significantly effecting its quality. The microstructure of T. castaneum was analyzed to reveal the mechanisms leading to death under microwave and heat treatments. Microwave radiation and conductive heating had different effects on the microstructure of the cuticle of adult T. castaneum and on the ultrastructure of the cells of the epidermis, fat body, and midgut. Both treatments caused a large cavity to appear in the nucleus and the disappearance of mitochondria and the Golgi apparatus. After microwave treatment, there was little change in the surface microstructure but the epidermis was of uneven thickness and the four outer layers of the cuticle were thinner. Nuclear size was essentially unchanged, but fat body cells were fewer and coalesced together. In contrast, conductive heating led to a disordered arrangement of cells on the surface of T. castaneum and indistinct boundaries between layers of the cuticle. The nuclei were enlarged and the fat body cells noticeably fewer and indistinct with a scattered distribution. Thus, microwave treatment produced less severe effects on the surface microstructure and cellular ultrastructure of T. castaneum than did conductive heating. It is concluded that these cellular and surface changes were responsible for the death of T. castaneum.

  16. Tribolium castaneum defensins are primarily active against Gram-positive bacteria.

    PubMed

    Tonk, Miray; Knorr, Eileen; Cabezas-Cruz, Alejandro; Valdés, James J; Kollewe, Christian; Vilcinskas, Andreas

    2015-11-01

    The red flour beetle Tribolium castaneum is a destructive insect pest of stored food and feed products, and a model organism for development, evolutionary biology and immunity. The insect innate immune system includes antimicrobial peptides (AMPs) with a wide spectrum of targets including viruses, bacteria, fungi and parasites. Defensins are an evolutionarily-conserved class of AMPs and a potential new source of antimicrobial agents. In this context, we report the antimicrobial activity, phylogenetic and structural properties of three T. castaneum defensins (Def1, Def2 and Def3) and their relevance in the immunity of T. castaneum against bacterial pathogens. All three recombinant defensins showed bactericidal activity against Micrococcus luteus and Bacillus thuringiensis serovar tolworthi, but only Def1 and Def2 showed a bacteriostatic effect against Staphylococcus epidermidis. None of the defensins showed activity against the Gram-negative bacteria Escherichia coli and Pseudomonas entomophila or against the yeast Saccharomyces cerevisiae. All three defensins were transcriptionally upregulated following a bacterial challenge, suggesting a key role in the immunity of T. castaneum against bacterial pathogens. Phylogenetic analysis showed that defensins from T. castaneum, mealworms, Udo longhorn beetle and houseflies cluster within a well-defined clade of insect defensins. We conclude that T. castaneum defensins are primarily active against Gram-positive bacteria and that other AMPs may play a more prominent role against Gram-negative species. PMID:26522790

  17. Structure of the Tribolium castaneum Telomerase Catalytic Subunit TERT

    SciTech Connect

    Gillis,A.; Schuller, A.; Skordalakes, E.

    2008-01-01

    A common hallmark of human cancers is the overexpression of telomerase, a ribonucleoprotein complex that is responsible for maintaining the length and integrity of chromosome ends. Telomere length deregulation and telomerase activation is an early, and perhaps necessary, step in cancer cell evolution. Here we present the high-resolution structure of the Tribolium castaneum catalytic subunit of telomerase, TERT. The protein consists of three highly conserved domains, organized into a ring-like structure that shares common features with retroviral reverse transcriptases, viral RNA polymerases and B-family DNA polymerases. Domain organization places motifs implicated in substrate binding and catalysis in the interior of the ring, which can accommodate seven to eight bases of double-stranded nucleic acid. Modelling of an RNA-DNA heteroduplex in the interior of this ring demonstrates a perfect fit between the protein and the nucleic acid substrate, and positions the 3'-end of the DNA primer at the active site of the enzyme, providing evidence for the formation of an active telomerase elongation complex.

  18. Oenocyte development in the red flour beetle Tribolium castaneum.

    PubMed

    Burns, Kevin A; Gutzwiller, Lisa M; Tomoyasu, Yoshinori; Gebelein, Brian

    2012-04-01

    Oenocytes are a specialized cell type required for lipid processing, pheromone secretion, and developmental signaling. Their development has been well characterized in Drosophila melanogaster, but it remains unknown whether the developmental program is conserved in other insect species. In this study, we compare and contrast the specification and development of larval oenocytes between Drosophila and the red flour beetle, Tribolium castaneum. First, we identify several useful reagents to label larval oenocytes, including both a Tribolium GFP enhancer trap line and a simple flurophore-conjugated streptavidin staining method that recognizes oenocytes across insect species. Second, we use these tools to describe oenocyte development in Tribolium embryos, and our findings provide evidence for conserved roles of MAP kinase signaling as well as the Spalt, Engrailed, hepatocyte nuclear factor-4, and ventral veins lacking factors in producing abdominal-specific oenocyte cells. However, Tribolium embryos produce four times as many oenocytes per abdominal segment as Drosophila, and unlike in Drosophila, these cells rapidly downregulate the expression of the Spalt transcription factor. Thus, these results provide new insight into the molecular pathways regulating oenocyte specification across insect species.

  19. Malpighian tubule development in the red flour beetle (Tribolium castaneum).

    PubMed

    King, Benedict; Denholm, Barry

    2014-11-01

    Malpighian tubules (MpTs) are the major organ for excretion and osmoregulation in most insects. MpT development is characterised for Drosophila melanogaster, but not other species. We therefore do not know the extent to which the MpT developmental programme is conserved across insects. To redress this we provide a comprehensive description of MpT development in the beetle Tribolium castaneum (Coleoptera), a species separated from Drosophila by >315 million years. We identify similarities with Drosophila MpT development including: 1) the onset of morphological development, beginning when tubules bud from the gut and proliferate to increase organ size. 2) the tubule is shaped by convergent-extension movements and oriented cell divisions. 3) differentiated tip cells activate EGF-signalling in distal MpT cells through the ligand Spitz. 4) MpTs contain two main cell types - principal and stellate cells, differing in morphology and gene expression. We also describe development of the beetle cryptonephridial system, an adaptation for water conservation, which represents a major modification of the MpT ground plan characterised by intimate association between MpTs and rectum. This work establishes a new model to compare MpT development across insects, and provides a framework to help understand how an evolutionary novelty - the cryptonephridial system - arose during organ evolution.

  20. Malpighian tubule development in the red flour beetle (Tribolium castaneum).

    PubMed

    King, Benedict; Denholm, Barry

    2014-11-01

    Malpighian tubules (MpTs) are the major organ for excretion and osmoregulation in most insects. MpT development is characterised for Drosophila melanogaster, but not other species. We therefore do not know the extent to which the MpT developmental programme is conserved across insects. To redress this we provide a comprehensive description of MpT development in the beetle Tribolium castaneum (Coleoptera), a species separated from Drosophila by >315 million years. We identify similarities with Drosophila MpT development including: 1) the onset of morphological development, beginning when tubules bud from the gut and proliferate to increase organ size. 2) the tubule is shaped by convergent-extension movements and oriented cell divisions. 3) differentiated tip cells activate EGF-signalling in distal MpT cells through the ligand Spitz. 4) MpTs contain two main cell types - principal and stellate cells, differing in morphology and gene expression. We also describe development of the beetle cryptonephridial system, an adaptation for water conservation, which represents a major modification of the MpT ground plan characterised by intimate association between MpTs and rectum. This work establishes a new model to compare MpT development across insects, and provides a framework to help understand how an evolutionary novelty - the cryptonephridial system - arose during organ evolution. PMID:25242057

  1. Neuropeptidome of Tribolium castaneum antennal lobes and mushroom bodies.

    PubMed

    Binzer, Marlene; Heuer, Carsten M; Kollmann, Martin; Kahnt, Jorg; Hauser, Frank; Grimmelikhuijzen, Cornelis J P; Schachtner, Joachim

    2014-02-01

    Neuropeptides are a highly diverse group of signaling molecules that affect a broad range of biological processes in insects, including development, metabolism, behavior, and reproduction. In the central nervous system, neuropeptides are usually considered to act as neuromodulators and cotransmitters that modify the effect of "classical" transmitters at the synapse. The present study analyzes the neuropeptide repertoire of higher cerebral neuropils in the brain of the red flour beetle Tribolium castaneum. We focus on two integrative neuropils of the olfactory pathway, the antennal lobes and the mushroom bodies. Using the technique of direct peptide profiling by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry, we demonstrate that these neuropils can be characterized by their specific neuropeptide expression profiles. Complementary immunohistological analyses of selected neuropeptides revealed neuropeptide distribution patterns within the antennal lobes and the mushroom bodies. Both approaches revealed consistent differences between the neuropils, underlining that direct peptide profiling by mass spectrometry is a fast and reliable method to identify neuropeptide content.

  2. Expression patterns of cysteine peptidase genes across the Tribolium castaneum life cycle provide clues to biological function

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The red flour beetle, Tribolium castaneum, is a major agricultural pest responsible for considerable loss of stored grain and cereal products worldwide. T. castaneum larvae have a highly compartmentalized gut, with cysteine peptidases mostly in the acidic anterior part of the midgut. We have descri...

  3. Residual activity of methoprene and novaluron as surface treatments to manage the flour beetles, Tribolium castaneum and Tribolium confusum

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The juvenile hormone analog methoprene and the chitin synthesis inhibitor novaluron were evaluated by exposing late-stage larvae of Tribolium castaneum (Herbst) or Tribolium confusum (Jacqueline DuVal), with food material on concrete, plywood, and floor tile. Larvae of T. castaneum were more suscept...

  4. Integrated analysis of cytochrome P450 gene superfamily in the red flour beetle, Tribolium castaneum

    PubMed Central

    2013-01-01

    Background The functional and evolutionary diversification of insect cytochrome P450s (CYPs) shaped the success of insects. CYPs constitute one of the largest and oldest gene superfamilies that are found in virtually all aerobic organisms. Because of the availability of whole genome sequence and well functioning RNA interference (RNAi), the red flour beetle, Tribolium castaneum serves as an ideal insect model for conducting functional genomics studies. Although several T. castaneum CYPs had been functionally investigated in our previous studies, the roles of the majority of CYPs remain largely unknown. Here, we comprehensively analyzed the phylogenetic relationship of all T. castaneum CYPs with genes in other insect species, investigated the CYP6BQ gene cluster organization, function and evolution, as well as examined the mitochondrial CYPs gene expression patterns and intron-exon organization. Results A total 143 CYPs were identified and classified into 26 families and 59 subfamilies. The phylogenetic trees of CYPs among insects across taxa provided evolutionary insight for the genetic distance and function. The percentage of singleton (33.3%) in T. castaneum CYPs is much less than those in Drosophila melanogaster (52.5%) and Bombyx mori (51.2%). Most members in the largest CYP6BQ gene cluster may make contribution to deltamethrin resistance in QTC279 strain. T. castaneum genome encodes nine mitochondrial CYPs, among them CYP12H1 is only expressed in the final instar larval stage. The intron-exon organizations of these mitochondrial CYPs are highly diverse. Conclusion Our studies provide a platform to understand the evolution and functions of T. castaneum CYP gene superfamily which will help reveal the strategies employed by insects to cope with their environment. PMID:23497158

  5. Characterization and expression of the beta-N-acetylglucosaminidase gene family of Tribolium castaneum

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Enzymes belonging to the Beta-N-acetylglucosaminidase (NAG) family cleave chitin oligosaccharides produced by the action of chitinases on chitin into the constituent N-acetylglucosamine monomer. Four genes encoding putative NAGs in the red flour beetle, Tribolium castaneum, namely TcNAG1, TcFDL, Tc...

  6. Mechanical properties of elytra from Tribolium castaneum wild-type and body color mutant strains

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Cuticle tanning in insects involves simultaneous cuticular hardening and pigmentation. The dynamic mechanical properties of the highly modified and cuticle-rich forewings (elytra) from Tribolium castaneum (red flour beetle) body color mutant strains were investigated to determine the relationship b...

  7. Tubulin superfamily genes in Tribolium castaneum, and use of a Tubulin promoter to drive transgene expression

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The use of native promoters to drive transgene expression has facilitated overexpression studies in Drosophila and other insects. We identified twelve Tubulin family members from the genome sequence of the red flour beetle, Tribolium castaneum, and used the promoter from one of these to drive cons...

  8. BeetleBase in 2010: Revisions to Provide Comprehensive Genomic Information for Tribolium castaneum

    Technology Transfer Automated Retrieval System (TEKTRAN)

    BeetleBase (http://www.beetlebase.org) has been updated to provide more comprehensive genomic information for the red flour beetle Tribolium castaneum. The database contains genomic sequence scaffolds mapped to 10 linkage groups (genome assembly release Tcas_3.0), genetic linkage maps, the official ...

  9. Dusky-like is required for epidermal pigmentation and metamorphosis in Tribolium castaneum

    PubMed Central

    Li, Chengjun; Yun, Xiaopei; Li, Bin

    2016-01-01

    Dusky-like (Dyl) is associated with the morphogenesis of embryonic denticle, adult sensory bristle and wing hair in Drosophila melanogaster. And whether Dyl involved in insect post-embryonic development and its signal transduction are poorly understood. Here, phylogenetic analysis revealed that dyl displayed one-to-one orthologous relationship among insects. In Tribolium castaneum, dyl is abundantly expressed at the late embryonic stage. Tissue-specific expression analysis at the late adult stage illustrated high expression of dyl in the fat body and ovary. Knockdown of dyl resulted in the defects in larval epidermal pigmentation and completely blocked the transitions from larval to pupal and pupal to adult stages of T. castaneum. We further discovered that dyl RNAi phenotypes were phenocopied by blimp-1 or shavenbaby (svb) silencing, and dyl was positively regulated by blimp-1 through svb in T. castaneum. These results suggest that Dyl functions downstream of Blimp-1 through Svb for larval epidermal pigmentation and metamorphosis. Moreover, ftz-f1 was down-regulated after RNA interference (RNAi) suppressing any of those three genes, indicating that Ftz-f1 works downstream of Dyl to mediate the effects of Blimp-1, Svb and Dyl on metamorphosis in T. castaneum. This study provides valuable insights into functions and signaling pathway of insect Dyl. PMID:26829909

  10. Chymotrypsin-like peptidases from Tribolium castaneum: A role in molting revealed by RNA interference

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Chymotrypsin-like peptidases (CTLPs) of insects are primarily secreted into the gut lumen where they act as digestive enzymes. We studied the gene family encoding CTLPs in the genome of the red flour beetle, Tribolium castaneum. Using an extended search pattern, we identified 14 TcCTLP genes that e...

  11. Evaluation of Quantitative PCR Reference Genes for Gene Expression Studies in Tribolium castaneum After Fungal Challenge

    Technology Transfer Automated Retrieval System (TEKTRAN)

    To investigate gene expression in Tribolium castaneum exposed to Beauveria bassiana, reference genes for qPCR were evaluated. Of these, the widely used genes for ß-actin, a-tubulin, and RPS6 were not stable. The most stable were ribosomal protein genes, RPS3, RPS18, and RPL13a. Syntaxin1, syntaxin6...

  12. Identification, cloning, and expression of a GHF9 cellulase from Tribolium castaneum (Coleoptera: Tenebrionidae)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The availability of sequenced insect genomes has allowed for discovery and functional characterization of novel genes and proteins. We report use of the Tribolium castaneum (Herbst) (red flour beetle) genome to identify, clone, express, and characterize a novel endo-ß-1,4-glucanase we named TcEG1 (...

  13. Mechanisms for horizontal transfer of methoprene from treated to untreated Tribolium castaneum (Herbst)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Experiments were performed to determine the relative impact of different mechanisms of horizontal transfer of methoprene by Tribolium castaneum (Herbst), the red flour beetle. Insects exposed to 5 methoprene treated developmental stages (late-stage larvae, pupae, or adults) resulted in 100% mortalit...

  14. Tcmof regulates larval/pupal development and female fecundity in red flour beetle, Tribolium castaneum.

    PubMed

    Wang, Yanyun; Li, Chengjun; Sang, Ming; Li, Bin

    2015-02-01

    Males absent on the first (MOF) was originally identified as an essential component of the X chromosome dosage compensation system in Drosophila melanogaster, and is also a member of the MYST family of histone acetyltransferases. MOF has been extensively studied in D. melanogaster and mammals. However, whether MOF is involved in dosage compensation and/or other vital functions for newly emerging model insects such as Tribolium castaneum, is unclear. We cloned the mof from T. castaneum, named Tcmof. Phylogenetic analysis revealed that mof is highly conserved in eukaryotes but lost in birds. qPCR showed that Tcmof was most highly expressed in the early embryo stage and equally expressed in males and females. Treating larvae with ds-Tcmof led 79.1% of the insects to arrest during its eclosion; the remaining insects died either in the larval stage or immediately following eclosion. Treating pupae with the same construct eliminated the fertility of T. castaneum. This effect was rescued by reciprocal crosses with wild-type females, but not males. We infer that the mof gene is essential for larval/pupal development and female fertility in T. castaneum.

  15. Methuselah-like genes affect development, stress resistance, lifespan and reproduction in Tribolium castaneum.

    PubMed

    Li, Chengjun; Zhang, Yi; Yun, Xiaopei; Wang, Yanyun; Sang, Ming; Liu, Xing; Hu, Xingxing; Li, Bin

    2014-10-01

    Methuselah (Mth) is associated with lifespan, stress resistance and reproduction in Drosophila melanogaster, but Mth is not present in nondrosophiline insects. A number of methuselah-likes (mthls) have been identified in nondrosophiline insects, but it is unknown whether the functions of mth are shared by mthls or are divergent from them. Five mthls have been identified in Tribolium castaneum. Although they have different developmental expression patterns, they all enhance resistance to starvation. Only mthl1 and mthl2 enhance resistance to high temperature, whereas mthl4 and mthl5 negatively regulate oxidative stress in T. castaneum. Unlike in the fly with mth mutation, knockdown of mthls, except mthl3, shortens the lifespan of T. castaneum. Moreover, mthl1 and mthl2 are critical for Tribolium development. mthl1 plays important roles in larval and pupal development and adult eclosion, while mthl2 is required for eclosion. Moreover, mthl1 and mthl2 silencing reduces the fertility of T. castaneum, and mthl1 and mthl4 are also essential for embryo development. In conclusion, mthls have a significant effect on insect development, lifespan, stress resistance and reproduction. These results provide experimental evidence for functional divergence among mthls/mth and clues for the signal transduction of Mthls.

  16. Retroactive maintains cuticle integrity by promoting the trafficking of Knickkopf into the procuticle of Tribolium castaneum

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Molting, or the replacement of the old exoskeleton with a new cuticle, is a complex developmental process that all insects must undergo to maintain growth. Our work has uncovered a major pathway involved in cuticular chitin maintenance wherein accumulation of the Tribolium castaneum knickkopf protei...

  17. Both UDP N-acetylglucosamine pyrophosphorylases of Tribolium castaneum are critical for molting, survival, and fecundity

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A bioinformatics search of the genome of the red flour beetle, Tribolium castaneum, resulted in the identification of two genes encoding proteins closely related to UDP-N-acetylglucosamine pyrophosphorylases (UAP), which provide the activated precursor, UDP-N-acetylglucosamine, for the synthesis of ...

  18. Susceptibility of Tribolium castaneum (Coleoptera: Tenebrionidae) and Trogoderma inclusum (Coleoptera: Dermestidae) to cold temperatures

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Studies were conducted by exposing different life stages of Tribolium castaneum (Herbst), the red flour beetle, and Trogoderma variabile (Ballion), the warehouse beetle, for different time intervals to -18°C. Assessments were made of direct mortality to eggs, larvae, and adults, and eventual adult e...

  19. Efficacy of aerosol applications of methoprene and synergized pyrethrin against Tribolium castaneum adults and eggs

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Experiments were performed to determine the efficacy of a single aerosol application of the insecticides methoprene and piperonyl butoxide-synergized pyrethrin, alone or in combination, and the insecticide carrier, Isopar M, against Tribolium castaneum (Herbst), the red flour beetle. The initial tes...

  20. Effects of methoprene and synergized pyrethrin aerosol applications on Tribolium castaneum (Herbst) populations

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Experiments were performed to investigate the effects of horizontal transfer of the insect growth regulator (IGR) methoprene on confined populations of Tribolium castaneum (Herbst) either with or without hidden refugia. Multiple applications were made with the IGR alone or combined with synergized p...

  1. Tribolium castaneum as a model for high-throughput RNAi screening.

    PubMed

    Knorr, Eileen; Bingsohn, Linda; Kanost, Michael R; Vilcinskas, Andreas

    2013-01-01

    Coleopteran insects are a highly diverse and successful order, and many beetle species are significant agricultural pests. New biorational strategies for managing populations of beetles and other insect species are needed as pests develop resistance to chemical insecticides and Bt toxins. There is now an opportunity to use genome sequence data to identify genes that are essential for insect growth, development, or survival as new targets for designing control technology. This goal requires a method for high-throughput in vivo screening of thousands of genes to identify candidate genes that, when their expression is disrupted, have a phenotype that may be useful in insect pest control. Tribolium castaneum, the red flour beetle, is a model organism that offers considerable advantages for such screening, including ease of rearing in large numbers, a sequenced genome, and a strong, systemic RNAi response for specific depletion of gene transcripts. The RNAi effect in T. castaneum can be elicited in any tissue and any stage by the injection of dsRNA into the hemocoel, and injection of dsRNA into adult females can even be used to identify phenotypes in offspring. A pilot RNAi screen (iBeetle) is underway. Several T. castaneum genes with promising RNAi phenotypes for further development as mechanisms for plant protection have been identified. These include heat shock protein 90, chitin synthase, the segmentation gene hairy, and a matrix metalloprotease. Candidate genes identified in T. castaneum screens can then be tested in agricultural pest species (in which screening is not feasible), to evaluate their effectiveness for use in potential plant-based RNAi control strategies. Delivery of dsRNA expressed by genetically modified crops to the midgut of phytophagous insects is under investigation as a new tool for very specific protection of plants from insect pest species. The T. castaneum screening platform offers a system for discovery of candidate genes with high potential

  2. Functional analysis of C1 family cysteine peptidases in the larval gut of Tenebrio molitor and Tribolium castaneum

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We studied protein digestion the tenebrionids Tenebrio molitor and Tribolium castaneum, pests of stored grains and grain products, to identify potential targets for biopesticide development. Tenebrionid larvae have highly compartmentalized guts, with primarily cysteine peptidases in the acidic anter...

  3. Glutathione S-transferase (GST) genes in the red flour beetle, Tribolium castaneum, and comparative analysis with five additional insects.

    PubMed

    Shi, Houxia; Pei, Lianghong; Gu, Shasha; Zhu, Shicheng; Wang, Yanyun; Zhang, Yi; Li, Bin

    2012-11-01

    Glutathione S-transferases are important detoxification enzymes involved in insecticide resistance. Sequencing the Tribolium castaneum genome provides an opportunity to investigate the structure, function, and evolution of GSTs on a genome-wide scale. Thirty-six putative cytosolic GSTs and 5 microsomal GSTs have been identified in T. castaneum. Furthermore, 40, 35, 13, 23, and 32 GSTs have been discovered the other insects, Drosophila, Anopheles, Apis, Bombyx, and Acyrthosiphon, respectively. Phylogenetic analyses reveal that insect-specific GSTs, Epsilon and Delta, are the largest species-specific expanded GSTs. In T. castaneum, most GSTs are tandemly arranged in three chromosomes. Particularly, Epsilon GSTs have an inverted long-fragment duplication in the genome. Other four widely distributed classes are highly conserved in all species. Given that GSTs specially expanded in Tribolium castaneum, these genes might help to resist poisonous chemical environments and produce resistance to kinds of different insecticides.

  4. Expression patterns of cysteine peptidase genes across the Tribolium castaneum life cycle provide clues to biological function.

    PubMed

    Perkin, Lindsey; Elpidina, Elena N; Oppert, Brenda

    2016-01-01

    The red flour beetle, Tribolium castaneum, is a major agricultural pest responsible for considerable loss of stored grain and cereal products worldwide. T. castaneum larvae have a highly compartmentalized gut, with cysteine peptidases mostly in the acidic anterior part of the midgut that are critical to the early stages of food digestion. In previous studies, we described 26 putative cysteine peptidase genes in T. castaneum (types B, L, O, F, and K) located mostly on chromosomes 3, 7, 8, and 10. In the present study, we hypothesized that specific cysteine peptidase genes could be associated with digestive functions for food processing based on comparison of gene expression profiles in different developmental stages, feeding and non-feeding. RNA-Seq was used to determine the relative expression of cysteine peptidase genes among four major developmental stages (egg, larvae, pupae, and adult) of T. castaneum. We also compared cysteine peptidase genes in T. castaneum to those in other model insects and coleopteran pests. By combining transcriptome expression, phylogenetic comparisons, response to dietary inhibitors, and other existing data, we identified key cysteine peptidases that T. castaneum larvae and adults use for food digestion, and thus new potential targets for biologically-based control products.

  5. Expression patterns of cysteine peptidase genes across the Tribolium castaneum life cycle provide clues to biological function

    PubMed Central

    Elpidina, Elena N.; Oppert, Brenda

    2016-01-01

    The red flour beetle, Tribolium castaneum, is a major agricultural pest responsible for considerable loss of stored grain and cereal products worldwide. T. castaneum larvae have a highly compartmentalized gut, with cysteine peptidases mostly in the acidic anterior part of the midgut that are critical to the early stages of food digestion. In previous studies, we described 26 putative cysteine peptidase genes in T. castaneum (types B, L, O, F, and K) located mostly on chromosomes 3, 7, 8, and 10. In the present study, we hypothesized that specific cysteine peptidase genes could be associated with digestive functions for food processing based on comparison of gene expression profiles in different developmental stages, feeding and non-feeding. RNA-Seq was used to determine the relative expression of cysteine peptidase genes among four major developmental stages (egg, larvae, pupae, and adult) of T. castaneum. We also compared cysteine peptidase genes in T. castaneum to those in other model insects and coleopteran pests. By combining transcriptome expression, phylogenetic comparisons, response to dietary inhibitors, and other existing data, we identified key cysteine peptidases that T. castaneum larvae and adults use for food digestion, and thus new potential targets for biologically-based control products. PMID:26819843

  6. The sperm ultrastructure and spermiogenesis of Tribolium castaneum (Coleoptera: Tenebrionidae) with evidence of cyst degeneration.

    PubMed

    Dias, Glenda; Lino-Neto, José; Mercati, David; Dallai, Romano

    2015-06-01

    Previous studies on the spermatogenesis of tenebrionid beetles showed the unusual formation of two antiparallel sperm bundles per cyst. In this work we reported this feature also in Tribolium castaneum using light and transmission electron microscopy. The sperm structure of T. castaneum, similar to other tenebrionids, consists of a three-layered acrosome, an elongated nucleus and a flagellum with a 9+9+2 axoneme, two accessory bodies and two asymmetric mitochondrial derivatives. The presence of two antiparallel sperm bundles per cyst also in Meloidae and Rhipiphoridae suggests that it is a strong trait synapomorphic for Tenebrionoidea. The huge degeneration of whole sperm cells in several cysts of testes during spermiogenesis is also described.

  7. Characterization of an allatotropin-like peptide receptor in the red flour beetle, Tribolium castaneum.

    PubMed

    Vuerinckx, Kristel; Verlinden, Heleen; Lindemans, Marleen; Broeck, Jozef Vanden; Huybrechts, Roger

    2011-10-01

    Following a reverse pharmacology approach, we identified an allatotropin-like peptide receptor in Tribolium castaneum. Allatotropins are multifunctional neuropeptides initially isolated from the tabacco hornworm, Manduca sexta. They have been shown to be myoactive, to be cardio-acceleratory, to inhibit active ion transport, to stimulate juvenile hormone production and release and to be involved in the photic entrainment of the circadian clock. A tissue distribution analysis of the T. castaneum allatotropin-like peptide receptor by means of qRT-PCR revealed a prominent sexual dimorphism, the transcript levels being significantly higher in the male fat body and reproductive system. The endogenous ligand of the receptor, Trica-ATL, is able to increase the frequency and tonus of contractions in the gut and in the reproductive tract of mature red flour beetles. PMID:21742031

  8. Susceptibility status of different life stages of Tribolium castaneum Herbst (Col: Tenebrionidae) to spinosad.

    PubMed

    Yousefnezhad-Irani, Roghayeh; Asghar, Pourmirza Ali

    2007-09-01

    Red flour beetle, Tribolium castaneum, is one the most important and worldwide pest of stored-products. To evaluate the susceptibility of different life stages of this pest we used a commercial formulation of biorational insecticide, Spinosad, Tracer against adults, young and old larvae and pupae using topical application bioassay method. Mortality was recorded after exclusion of pupae and 24, 48 and 72 h posttreatment for the other life stages. Spinosad did not cause pupal mortality. Comparison of LD50 values of different life stages of T. castaneum at 48 h exposure-time revealed that, young larvae and adults were more susceptible to spinosad than the old larvae. Overlapping of 95% CL revealed similarity of susceptibility of young larvae and adult insects to spinosad. Due to compatibility of spinosad with environment, this compound could be considered as a useful tool in the control of the pest in question.

  9. The sperm ultrastructure and spermiogenesis of Tribolium castaneum (Coleoptera: Tenebrionidae) with evidence of cyst degeneration.

    PubMed

    Dias, Glenda; Lino-Neto, José; Mercati, David; Dallai, Romano

    2015-06-01

    Previous studies on the spermatogenesis of tenebrionid beetles showed the unusual formation of two antiparallel sperm bundles per cyst. In this work we reported this feature also in Tribolium castaneum using light and transmission electron microscopy. The sperm structure of T. castaneum, similar to other tenebrionids, consists of a three-layered acrosome, an elongated nucleus and a flagellum with a 9+9+2 axoneme, two accessory bodies and two asymmetric mitochondrial derivatives. The presence of two antiparallel sperm bundles per cyst also in Meloidae and Rhipiphoridae suggests that it is a strong trait synapomorphic for Tenebrionoidea. The huge degeneration of whole sperm cells in several cysts of testes during spermiogenesis is also described. PMID:25867758

  10. Phosphine resistance in Tribolium castaneum and Rhyzopertha dominica from stored wheat in Oklahoma.

    PubMed

    Opit, G P; Phillips, T W; Aikins, M J; Hasan, M M

    2012-08-01

    Phosphine gas, or hydrogen phosphide (PH3), is the most common insecticide applied to durable stored products worldwide and is routinely used in the United States for treatment of bulk-stored cereal grains and other durable stored products. Research from the late 1980s revealed low frequencies of resistance to various residual grain protectant insecticides and to phosphine in grain insect species collected in Oklahoma. The present work, which used the same previously established discriminating dose bioassays for phosphine toxicity as in the earlier study, evaluated adults of nine different populations of red flour beetle, Tribolium castaneum (Herbst), and five populations of lesser grain borer, Rhyzopertha dominica (F.) collected from different geographic locations in Oklahoma. One additional population for each species was a laboratory susceptible strain. Discriminating dose assays determined eight out of the nine T. castaneum populations, and all five populations of R. dominica, contained phosphine-resistant individuals, and highest resistance frequencies were 94 and 98%, respectively. Dose-response bioassays and logit analyses determined that LC99 values were approximately 3 ppm for susceptible and 377 ppm for resistant T. castaneum, and approximately 2 ppm for susceptible and 3,430 ppm for resistant R. dominica. The most resistant T. castaneum population was 119-fold more resistant than the susceptible strain and the most resistant R. dominica population was over 1,500-fold more resistant. Results suggest a substantial increase in phosphine resistance in these major stored-wheat pests in the past 21 yr, and these levels of resistance to phosphine approach those reported for other stored-grain pest species in other countries.

  11. Concentration dependent toxicokinetics of copper in the red flour beetle Tribolium castaneum (Coleoptera: Tenebrionidae).

    PubMed

    Bednarska, Agnieszka J; Stępień, Katarzyna

    2015-11-01

    To predict internal metal concentrations in animals under specific environmental exposures, the relationship between the exposure concentrations and values of toxicokinetic parameters must be known. At high exposure levels, the availability of carriers transporting metal ions through cellular membranes may become limited, thereby decreasing the assimilation rates (k A ). Furthermore, increased metal concentrations in food may result in greater damage to the gut and reduce the assimilation efficiency and/or increase the elimination rate (k E ). Therefore, k A should decrease and k E should increase with increasing metal concentrations. In fact, our study on Tribolium castaneum exposed to Cu at 500, 1000, 2000 and 4000 mg kg(-1) of dry flour showed that with increasing Cu concentrations, k A decreased from 0.0042 day(-1) at 500 mg kg(-1) to 0.0026 day(-1) at 4000 mg kg(-1) in females and from 0.0029 to 0.001 day(-1) in males and k E increased from 0.027 to 0.064 day(-1) and from 0.018 to 0.04 day(-1) in females and males, respectively. Significant differences in k A between the sexes were observed at 2000 and 4000 mg kg(-1), whereas significant differences between treatments were found for k A in males. Copper was efficiently regulated by T. castaneum: an eightfold increase in exposure concentrations resulted in only a ca. twofold increase in the internal concentration. No Cu effect on the respiratory metabolism of T. castaneum was found.

  12. Repellent Constituents of essential oil from Citrus wilsonii stem barks against Tribolium castaneum.

    PubMed

    Wua, Yan; Chenb, Hai-Ping; Wei, Jian-Yu; Yang, Kai; Tian, Zhao-Fu; Li, Xiao-Lan; Wang, Ping-Juan; Wang, Cheng-Fang; Du, Shu-Shan; Cai, Qian

    2014-10-01

    The essential oil obtained from Citrus wilsonii Tanaka stem barks with hydrodistillation was investigated by GC-FID and GC-MS. The main components of the essential oil were identified to be nerol acetate (44.5%), nerol (13.6%), citronellyl propionate (13.5%) and α-terpineol (3.6%). Among them, the four active constituents, predicted with a bioactivity-test, were isolated and identified as nerolacetate, nerol, citronellyl propionate and α-terpineol. It was found that the essential oil of C. wilsonii stem barks possessed strong repellency (86% and 92%, respectively, at 78.6 nL/cm2, after 2 and 4 h treatment) against Tribolium castaneum adults. Repellency of the four active compounds was also determined. Nerolacetate, nerol, citronellyl propionate and α-terpineol were strongly repellent (100%, 100%, 90% and 96%, respectively, at 15.7 nL/cm2, after 2h treatment) against T. castaneum. Nerol exhibited the same level of repellency as the positive control, DEET. The results indicate that the essential oil of C. wilsonii stem barks and its active compounds have the potential to be developed as natural repellents for control of T. castaneum.

  13. JH modulates a cellular immunity of Tribolium castaneum in a Met-independent manner.

    PubMed

    Hepat, Rahul; Kim, Yonggyun

    2014-04-01

    Juvenile hormone (JH) regulates diverse physiological processes in insects during entire developmental stages. Especially, the identification of Methoprene-tolerant (Met), a JH nuclear receptor, allows us to better understand molecular actions of JH to control gene expressions related with metamorphosis. However, several physiological processes including cellular immune response and some molecular actions of JH have been suspected to be mediated via its non-genomic actions. To prove its non-genomic action, JH nuclear signals were suppressed by RNA interference (RNAi) of Met or its downstream gene, Krüppel homolog 1 (Kr-h1), in the red flour beetle, Tribolium castaneum. These RNAi-treated larvae failed to undergo a normal development and suffered precocious metamorphosis. Hemocytes of T. castaneum exhibited their spreading behavior on extracellular matrix and nodule formation in response to bacterial challenge. When the larvae were treated with either RNAi of Met or Kr-h1, the hemocytes of the treated larvae were responsive to JH without any significant difference with those of control larvae. These results suggest that the response of hemocytes to JH is not mediated by its nuclear signal. On the other hand, the JH modulation of hemocyte behaviors of T. castaneum was significantly influenced by membrane and cytosolic protein activities, in which ethoxyzolamide (a specific inhibitor of carbonic anhydrase), calphostin C (a specific inhibitor of protein kinase C) or ouabain (a specific inhibitor of Na(+)-K(+) ATPase) significantly suppressed the responsiveness of hemocytes to JH.

  14. First evidence of DNA methylation in insect Tribolium castaneum: environmental regulation of DNA methylation within heterochromatin.

    PubMed

    Feliciello, Isidoro; Parazajder, Josip; Akrap, Ivana; Ugarković, Durđica

    2013-05-01

    DNA methylation has been studied in many eukaryotic organisms, in particular vertebrates, and was implicated in developmental and phenotypic variations. Little is known about the role of DNA methylation in invertebrates, although insects are considered as excellent models for studying the evolution of DNA methylation. In the red flour beetle, Tribolium castaneum (Tenebrionidae, Coleoptera), no evidence of DNA methylation has been found till now. In this paper, a cytosine methylation in Tribolium castaneum embryos was detected by methylation sensitive restriction endonucleases and immuno-dot blot assay. DNA methylation in embryos is followed by a global demethylation in larvae, pupae and adults. DNA demethylation seems to proceed actively through 5-hydroxymethylcytosine, most probably by the action of TET enzyme. Bisulfite sequencing of a highly abundant satellite DNA located in pericentromeric heterochromatin revealed similar profile of cytosine methylation in adults and embryos. Cytosine methylation was not only restricted to CpG sites but was found at CpA, CpT and CpC sites. In addition, complete cytosine demethylation of heterochromatic satellite DNA was induced by heat stress. The results reveal existence of DNA methylation cycling in T. castaneum ranging from strong overall cytosine methylation in embryos to a weak DNA methylation in other developmental stages. Nevertheless, DNA methylation is preserved within heterochromatin during development, indicating its role in heterochromatin formation and maintenance. It is, however, strongly affected by heat stress, suggesting a role for DNA methylation in heterochromatin structure modulation during heat stress response.

  15. iBeetle-Base: a database for RNAi phenotypes in the red flour beetle Tribolium castaneum.

    PubMed

    Dönitz, Jürgen; Schmitt-Engel, Christian; Grossmann, Daniela; Gerischer, Lizzy; Tech, Maike; Schoppmeier, Michael; Klingler, Martin; Bucher, Gregor

    2015-01-01

    The iBeetle-Base (http://ibeetle-base.uni-goettingen.de) makes available annotations of RNAi phenotypes, which were gathered in a large scale RNAi screen in the red flour beetle Tribolium castaneum (iBeetle screen). In addition, it provides access to sequence information and links for all Tribolium castaneum genes. The iBeetle-Base contains the annotations of phenotypes of several thousands of genes knocked down during embryonic and metamorphic epidermis and muscle development in addition to phenotypes linked to oogenesis and stink gland biology. The phenotypes are described according to the EQM (entity, quality, modifier) system using controlled vocabularies and the Tribolium morphological ontology (TrOn). Furthermore, images linked to the respective annotations are provided. The data are searchable either for specific phenotypes using a complex 'search for morphological defects' or a 'quick search' for gene names and IDs. The red flour beetle Tribolium castaneum has become an important model system for insect functional genetics and is a representative of the most species rich taxon, the Coleoptera, which comprise several devastating pests. It is used for studying insect typical development, the evolution of development and for research on metabolism and pest control. Besides Drosophila, Tribolium is the first insect model organism where large scale unbiased screens have been performed.

  16. Repellent Constituents of essential oil from Citrus wilsonii stem barks against Tribolium castaneum.

    PubMed

    Wua, Yan; Chenb, Hai-Ping; Wei, Jian-Yu; Yang, Kai; Tian, Zhao-Fu; Li, Xiao-Lan; Wang, Ping-Juan; Wang, Cheng-Fang; Du, Shu-Shan; Cai, Qian

    2014-10-01

    The essential oil obtained from Citrus wilsonii Tanaka stem barks with hydrodistillation was investigated by GC-FID and GC-MS. The main components of the essential oil were identified to be nerol acetate (44.5%), nerol (13.6%), citronellyl propionate (13.5%) and α-terpineol (3.6%). Among them, the four active constituents, predicted with a bioactivity-test, were isolated and identified as nerolacetate, nerol, citronellyl propionate and α-terpineol. It was found that the essential oil of C. wilsonii stem barks possessed strong repellency (86% and 92%, respectively, at 78.6 nL/cm2, after 2 and 4 h treatment) against Tribolium castaneum adults. Repellency of the four active compounds was also determined. Nerolacetate, nerol, citronellyl propionate and α-terpineol were strongly repellent (100%, 100%, 90% and 96%, respectively, at 15.7 nL/cm2, after 2h treatment) against T. castaneum. Nerol exhibited the same level of repellency as the positive control, DEET. The results indicate that the essential oil of C. wilsonii stem barks and its active compounds have the potential to be developed as natural repellents for control of T. castaneum. PMID:25522550

  17. Repellent and fumigant toxicity of essential oil from Thymus persicus against Tribolium castaneum and Callosobruchus maculatus.

    PubMed

    Moharramipour, S; Taghizadeh, A; Meshkatalsadat, M H; Talebi, A A; Fathipour, Y

    2008-01-01

    Repellent and insecticidal activity of the essential oil extracted from Thymus persicus (Roniger ex Reach. F.) Jalas was evaluated against two stored-product beetles Tribolium castaneum (Herbst) and Callosobruchus maculatus (F.). Dry flowering aerial parts of the plant were subjected to hydro distillation using a modified Clevenger-type apparatus. The repellent and fumigant toxicity were tested against 1-7 days old adult beetles at 27 +/- 1 degrees C and 65 +/- 5% RH in dark condition. The repellency on C. maculatus and T. castaneum at highest concentration (2 microL/mL acetone) was 82.40% and 70.40% respectively. Fumigation bioassays showed that C. maculatus adults were significantly more susceptible (LC50 = 2.39 microL/L air) to the essential oil than T. castaneum adults (LC50 = 234.42 microL/L air). It could be concluded that T. persicus may have potential for applications in management of stored-product pests because of its safety, strong repellency and fumigant toxicity. PMID:19226805

  18. Microbiota Plays a Role in Oral Immune Priming in Tribolium castaneum

    PubMed Central

    Futo, Momir; Armitage, Sophie A. O.; Kurtz, Joachim

    2016-01-01

    Animals are inhabited by a diverse community of microorganisms. The relevance of such microbiota is increasingly being recognized across a broad spectrum of species, ranging from sponges to primates, revealing various beneficial roles that microbes can play. The red flour beetle Tribolium castaneum represents a well-established experimental model organism for studying questions in ecology and evolution, however, the relevance of its microbial community is still largely unknown. T. castaneum larvae orally exposed to bacterial components of the entomopathogen Bacillus thuringiensis bv. tenebrionis showed increased survival upon a subsequent challenge with spores of this bacterium. To investigate whether T. castaneum microbiota plays a role in this phenomenon, we established a protocol for raising microbe-free larvae and subsequently tested whether they differ in their ability to mount such a priming response. Here we demonstrate that larvae with significantly lowered microbial loads, show decreased survival upon secondary challenge with B. thuringiensis bv. tenebrionis spores, compared to animals that were allowed to regain their microbiota before priming. Although the exact mechanism of oral immune priming is unclear, we here suggest that microbiota plays a crucial role in oral immune priming in this species. PMID:26779124

  19. Microbiota Plays a Role in Oral Immune Priming in Tribolium castaneum.

    PubMed

    Futo, Momir; Armitage, Sophie A O; Kurtz, Joachim

    2015-01-01

    Animals are inhabited by a diverse community of microorganisms. The relevance of such microbiota is increasingly being recognized across a broad spectrum of species, ranging from sponges to primates, revealing various beneficial roles that microbes can play. The red flour beetle Tribolium castaneum represents a well-established experimental model organism for studying questions in ecology and evolution, however, the relevance of its microbial community is still largely unknown. T. castaneum larvae orally exposed to bacterial components of the entomopathogen Bacillus thuringiensis bv. tenebrionis showed increased survival upon a subsequent challenge with spores of this bacterium. To investigate whether T. castaneum microbiota plays a role in this phenomenon, we established a protocol for raising microbe-free larvae and subsequently tested whether they differ in their ability to mount such a priming response. Here we demonstrate that larvae with significantly lowered microbial loads, show decreased survival upon secondary challenge with B. thuringiensis bv. tenebrionis spores, compared to animals that were allowed to regain their microbiota before priming. Although the exact mechanism of oral immune priming is unclear, we here suggest that microbiota plays a crucial role in oral immune priming in this species. PMID:26779124

  20. Next generation sequencing based transcriptome analysis of septic-injury responsive genes in the beetle Tribolium castaneum.

    PubMed

    Altincicek, Boran; Elashry, Abdelnaser; Guz, Nurper; Grundler, Florian M W; Vilcinskas, Andreas; Dehne, Heinz-Wilhelm

    2013-01-01

    Beetles (Coleoptera) are the most diverse animal group on earth and interact with numerous symbiotic or pathogenic microbes in their environments. The red flour beetle Tribolium castaneum is a genetically tractable model beetle species and its whole genome sequence has recently been determined. To advance our understanding of the molecular basis of beetle immunity here we analyzed the whole transcriptome of T. castaneum by high-throughput next generation sequencing technology. Here, we demonstrate that the Illumina/Solexa sequencing approach of cDNA samples from T. castaneum including over 9.7 million reads with 72 base pairs (bp) length (approximately 700 million bp sequence information with about 30× transcriptome coverage) confirms the expression of most predicted genes and enabled subsequent qualitative and quantitative transcriptome analysis. This approach recapitulates our recent quantitative real-time PCR studies of immune-challenged and naïve T. castaneum beetles, validating our approach. Furthermore, this sequencing analysis resulted in the identification of 73 differentially expressed genes upon immune-challenge with statistical significance by comparing expression data to calculated values derived by fitting to generalized linear models. We identified up regulation of diverse immune-related genes (e.g. Toll receptor, serine proteinases, DOPA decarboxylase and thaumatin) and of numerous genes encoding proteins with yet unknown functions. Of note, septic-injury resulted also in the elevated expression of genes encoding heat-shock proteins or cytochrome P450s supporting the view that there is crosstalk between immune and stress responses in T. castaneum. The present study provides a first comprehensive overview of septic-injury responsive genes in T. castaneum beetles. Identified genes advance our understanding of T. castaneum specific gene expression alteration upon immune-challenge in particular and may help to understand beetle immunity in general.

  1. Next generation sequencing based transcriptome analysis of septic-injury responsive genes in the beetle Tribolium castaneum.

    PubMed

    Altincicek, Boran; Elashry, Abdelnaser; Guz, Nurper; Grundler, Florian M W; Vilcinskas, Andreas; Dehne, Heinz-Wilhelm

    2013-01-01

    Beetles (Coleoptera) are the most diverse animal group on earth and interact with numerous symbiotic or pathogenic microbes in their environments. The red flour beetle Tribolium castaneum is a genetically tractable model beetle species and its whole genome sequence has recently been determined. To advance our understanding of the molecular basis of beetle immunity here we analyzed the whole transcriptome of T. castaneum by high-throughput next generation sequencing technology. Here, we demonstrate that the Illumina/Solexa sequencing approach of cDNA samples from T. castaneum including over 9.7 million reads with 72 base pairs (bp) length (approximately 700 million bp sequence information with about 30× transcriptome coverage) confirms the expression of most predicted genes and enabled subsequent qualitative and quantitative transcriptome analysis. This approach recapitulates our recent quantitative real-time PCR studies of immune-challenged and naïve T. castaneum beetles, validating our approach. Furthermore, this sequencing analysis resulted in the identification of 73 differentially expressed genes upon immune-challenge with statistical significance by comparing expression data to calculated values derived by fitting to generalized linear models. We identified up regulation of diverse immune-related genes (e.g. Toll receptor, serine proteinases, DOPA decarboxylase and thaumatin) and of numerous genes encoding proteins with yet unknown functions. Of note, septic-injury resulted also in the elevated expression of genes encoding heat-shock proteins or cytochrome P450s supporting the view that there is crosstalk between immune and stress responses in T. castaneum. The present study provides a first comprehensive overview of septic-injury responsive genes in T. castaneum beetles. Identified genes advance our understanding of T. castaneum specific gene expression alteration upon immune-challenge in particular and may help to understand beetle immunity in general

  2. Characterisation and tissue distribution of the PISCF allatostatin receptor in the red flour beetle, Tribolium castaneum.

    PubMed

    Audsley, Neil; Vandersmissen, Hans Peter; Weaver, Robert; Dani, Paulina; Matthews, June; Down, Rachel; Vuerinckx, Kristel; Kim, Young-Joon; Vanden Broeck, Jozef

    2013-01-01

    The insect PISCF/allatostatins (ASTs) are pleiotropic peptides that are involved in the regulation of juvenile hormone biosynthesis, are myoinhibitory on the gut and the heart, and suppress feeding in various insects, but their roles in beetles are poorly understood. To provide further insight into the significance of PISCF/ASTs in beetles, the PISCF/AST receptor from Tribolium castaneum has been characterised and its tissue distribution determined. The biological activity of the T. castaneum PISCF/AST (Trica-AS) was also investigated. The Trica-AS receptor shows high sequence homology to other insect PISCF/AST receptors, which are related to the mammalian somatostatin/opioid receptors, a family of G protein-coupled receptors. The Trica-AS receptor was activated in a dose-dependent manner by both Trica-AS and T. castaneum allatostatin double C (Trica-ASTCC) as well as Manduca sexta-allatostatin (Manse-AS). Other allatoregulatory peptides (a FLG/AST, a MIP/AST and an allatotropin) and somatostatin(14) were inactive on this receptor. Receptor transcript levels in tissues, determined by qRT-PCR, were highest in the head and the gut, with variable amounts in the fat body and reproductive organs. There were measurable differences in receptor levels of the head, fat body and reproductive organs between males and females. There was also a widespread distribution of Trica-AS in various tissues of T. castaneum. The Trica-AS peptide precursor was most abundant in the head and there was a significant difference between levels in the heads and reproductive organs of males and females. Whole mount immunocytochemistry localised Trica-AS in the median and lateral neurosecretory cells of the brain, in the corpus cardiacum and throughout the ventral nerve cord. The peptide was also present in midgut neurosecretory cells, but no immunostaining was detected in the reproductive organs or Malpighian tubules. The widespread distribution of both Trica-AS and its receptor suggest this

  3. Capture of Tribolium castaneum and Tribolium confusum (Coleoptera: Tenebrionidae) in Floor Traps: The Effect of Previous Captures.

    PubMed

    Athanassiou, Christos G; Kavallieratos, Nickolas G; Campbell, James F

    2016-02-01

    The impact of prior captures on the trapping performance of floor traps was evaluated for the red flour beetle, Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae), and the confused flour beetle, Tribolium confusum Jacquelin du Val (Coleoptera: Tenebrionidae), in laboratory conditions. The effect of trap seeding, adding adults of the same or different species, was evaluated in order to determine possible effects of prior captures in the trap on each species' behavioral responses. The presence of seeded beetles of the same species resulted in an increase in beetle captures for both T. castaneum and T. confusum, but when traps were seeded with the opposite species, there was no increase in beetle captures for either species, and for T. castaneum overall captures in both seeded and unseeded traps was reduced. Overall, T. castaneum tended to have greater captures than T. confusum regardless of the treatment. When the two species were released together, this negated the increased response to seeded traps observed in the single-species treatments. These findings suggest the potential that the presence of beetles in a trap may be influencing the response of beetles in a nearby trap and that T. castaneum and T. confusum when they occur together may influence each other's response to traps.

  4. Tribolium castaneum Apolipophorin-III acts as an immune response protein against Bacillus thuringiensis Cry3Ba toxic activity.

    PubMed

    Contreras, Estefanía; Rausell, Carolina; Real, M Dolores

    2013-07-01

    In this study, a 2.1-fold Apolipophorin-III mRNA up-regulation was found in Tribolium castaneum larvae challenged with Bacillus thuringiensis Cry3Ba spore-crystal mixture. Knockdown of Apolipophorin-III by RNAi resulted in increased T. castaneum larvae susceptibility following Cry3Ba spore-crystal treatment, demonstrating Apolipophorin-III involvement in insect defense against B. thuringiensis. We showed that Apolipophorin-III participates in T. castaneum immune response to B. thuringiensis activating the prophenoloxidase cascade since: (i) phenoloxidase activity significantly increased after Cry3Ba spore-crystal treatment compared to untreated or Cry1Ac spore-crystal treated larvae and (ii) phenoloxidase activity in Cry3Ba spore-crystal treated Apolipophorin-III silenced larvae was 71±14% lower than that of non-silenced intoxicated larvae.

  5. Models to predict mortality of Tribolium castaneum (Coleoptera: Tenebrionidae) exposed to elevated temperatures during structural heat treatments.

    PubMed

    Jian, Fuji; Subramanyam, Bhadriraju; Jayas, Digvir S; White, Noel D G

    2013-10-01

    Novel thermal death models were developed with certain assumptions, and these models were validated by using actual heat treatment data collected under laboratory conditions at constant temperatures over time and in commercial food-processing facilities where temperatures were dynamically changing over time. The predicted mortalities of both young larvae and adults of the red flour beetle, Tribolium castaneum (Herbst), were within 92-99% of actual measured insect mortalities. There was good concordance between predicted and observed mortalities of young larvae and adults of T. castaneum exposed to constant temperatures in laboratory growth chambers and at variable temperatures during structural heat treatments of commercial food-processing facilities. The models developed in this study can be used to determine effectiveness of structural heat treatments in killing young larvae and adults of T. castaneum and for characterizing insect thermotolerance.

  6. Evaluation of reference genes for quantitative polymerase chain reaction across life cycle stages and tissue types of Tribolium castaneum.

    PubMed

    Toutges, Michelle J; Hartzer, Kris; Lord, Jeffrey; Oppert, Brenda

    2010-08-25

    The genome of the genetic model for coleopteran insects, Tribolium castaneum, is now available for downstream applications. To facilitate gene expression studies in T. castaneum, genes were evaluated for suitability as normalizers in comparisons across tissues and/or developmental stages. In less diverse samples, such as comparisons within developmental stages or tissue only, normalizers for mRNA were more stable and consistent. Overall, the genes for ribosomal proteins rps6, rpl13a, rps3, and rps18 were the most stable normalizers for broad scale gene expression analysis in T. castaneum. However, their stability ranking was dependent upon the instrument as well as the analysis program. These data emphasize the need to optimize normalizers used in all real-time polymerase chain reaction experiments specifically for the experimental conditions and thermocycler and to carefully evaluate data generated by computational algorithms.

  7. A cell line derived from the red flour beetle Tribolium castaneum (Coleoptera: Tenebrionidae).

    PubMed

    Goodman, Cynthia L; Stanley, David; Ringbauer, Joseph A; Beeman, Richard W; Silver, Kristopher; Park, Yoonseong

    2012-08-01

    The red flour beetle, Tribolium castaneum, is a model organism for agricultural and medical research and its complete genome is sequenced. We established a continuously replicating T. castaneum cell line to complement existing physiological, genetic, and genomic research tools. We set up trial cell cultures from egg, pupa, and adult stages as tissue sources and incubated them in six separate cell culture media to determine the optimal combination of tissue source and medium for cell replication. Our most promising culture was generated by co-culturing adult (∼75 %) and pupal tissues in EX-CELL 420 medium containing 9 % FBS. Our new cell culture is designated BCIRL-TcA-CLG1 (TcA) and it has been subcultured more than 90 times. Amplification of genomic DNA with species-specific primers yielded DNA fragments of the expected sizes and with sequences identical to those from the published Tribolium genome. Additionally, we characterized this line using DNA fingerprinting (DAF-PCR) and compared it with three other coleopteran cell lines and its conspecific pupae to confirm identity. Its doubling time is 155.2 hr. Early passages consisted of attached cells and vesicles in suspension, whereas later passages consisted primarily of attached, spherical cells. Similar to other established cell lines, the ploidy of TcA cells was variable, ranging from 20 chromosomes/cell (diploid) to above 30 chromosomes/cell. TcA cells withstood incubation at 40°C for 1 h with no decrease in viability. We recorded increased levels of one heat shock protein (43 kDa) and of the hsp68a transcript following exposure to 40°C. Taken together, this represents the first report of a continuously replicating T. castaneum cell line. We expect the BCIRL-TcA-CLG1 line will become a useful tool in Tribolium research.

  8. Transovarial Effect of Novaluron on Tribolium castaneum (Coleoptera: Tenebrionidae) After Termination of Direct Contact.

    PubMed

    Trostanetsky, A; Kostyukovsky, M; Quinn, E

    2015-01-01

    The insect growth regulator novaluron (Rimon 10 EC, Makhteshim-Agan Ltd, Israel) is used against many field pests on corn, vegetables, orchards, forests, and cotton plantations. Previously, we studied various effects of novaluron on stored grain pests. Termination in Tribolium castaneum (Herbst) eggs hatching after treating adults with novaluron and following restoration after adult transfer to untreated media was observed. The objective of this study was to investigate the restoration of T. castaneum egg hatch following transfer of adults from treated media to untreated favorable and unfavorable media. The time needed for hatching restoration of 50% of eggs laid by adults transferred from novaluron (1 ppm) treated flour to untreated flour (RT50) was 2.7 d. RT50 for those transferred to untreated wheat grain was 4.1 d. RT90 in flour was 3.6 d, in grain--6.1 d. Varieties of RTs in grain and in flour with nonoverlapping confidence intervals indicate that RTs were significantly different. Delay of eggs hatching restoration for adults transferred from treated flour to unfavorable media (Petri dishes with limited amount of flour, lying of eggs not detected) was observed. RT50 in flour was 2.1 d and RT90--3.1 d, while RT50 in the unfavorable media was 3.4 d and RT90 6.5 d. Delayed effect of egg hatching restoration after adult transfer to unfavorable media provides evidence of the significant role of insect physiological state in novaluron excretion and (or) degradation by T. castaneum females.

  9. Transovarial Effect of Novaluron on Tribolium castaneum (Coleoptera: Tenebrionidae) After Termination of Direct Contact

    PubMed Central

    Trostanetsky, A.; Kostyukovsky, M.; Quinn, E.

    2015-01-01

    The insect growth regulator novaluron (Rimon 10 EC, Makhteshim-Agan Ltd, Israel) is used against many field pests on corn, vegetables, orchards, forests, and cotton plantations. Previously, we studied various effects of novaluron on stored grain pests. Termination in Tribolium castaneum (Herbst) eggs hatching after treating adults with novaluron and following restoration after adult transfer to untreated media was observed. The objective of this study was to investigate the restoration of T. castaneum egg hatch following transfer of adults from treated media to untreated favorable and unfavorable media. The time needed for hatching restoration of 50% of eggs laid by adults transferred from novaluron (1 ppm) treated flour to untreated flour (RT50) was 2.7 d. RT50 for those transferred to untreated wheat grain was 4.1 d. RT90 in flour was 3.6 d, in grain—6.1 d. Varieties of RTs in grain and in flour with nonoverlapping confidence intervals indicate that RTs were significantly different. Delay of eggs hatching restoration for adults transferred from treated flour to unfavorable media (Petri dishes with limited amount of flour, lying of eggs not detected) was observed. RT50 in flour was 2.1 d and RT90—3.1 d, while RT50 in the unfavorable media was 3.4 d and RT90 6.5 d. Delayed effect of egg hatching restoration after adult transfer to unfavorable media provides evidence of the significant role of insect physiological state in novaluron excretion and (or) degradation by T. castaneum females. PMID:26363061

  10. Structure and function of the homeotic gene complex (HOM-C) in the beetle, Tribolium castaneum

    NASA Technical Reports Server (NTRS)

    Beeman, R. W.; Stuart, J. J.; Brown, S. J.; Denell, R. E.; Spooner, B. S. (Principal Investigator)

    1993-01-01

    The powerful combination of genetic, developmental and molecular approaches possible with the fruit fly, Drosophila melanogaster, has led to a profound understanding of the genetic control of early developmental events. However, Drosophila is a highly specialized long germ insect, and the mechanisms controlling its early development may not be typical of insects or Arthropods in general. The beetle, Tribolium castaneum, offers a similar opportunity to integrate high resolution genetic analysis with the developmental/molecular approaches currently used in other organisms. Early results document significant differences between insect orders in the functions of genes responsible for establishing developmental commitments.

  11. Effect of cold water extracts of Acacia modesta Wall. and Glycyrrhiza glabra Linn. on Tribolium castaneum and Lemna minor.

    PubMed

    Nazeefullah, Sayed; Dastagir, Ghulam; Ahmad, Bashir

    2014-03-01

    The aim of the present study was to introduce an alternative way for insects control through biodegradable plants materials. The different cold water extracts dilutions of Acacia modesta and Glycyrrhiza glabra were tested against Tribolium castaneum. The extracts dilutions of both plants caused mortality of the Tribolium castaneum. ANOVA revealed that dilutions and plants were highly significant. The interaction between plants and dilutions was also significant at P < 0.05. Phytotoxic activity showed that dilutions of Acacia modesta and Glycyrrhiza glabra extracts significantly inhibited the growth of Lemna minor. ANOVA showed that dilutions of both plants extracts were significant at P < 0.05.

  12. Cannibalism facilitates the use of a novel environment in the flour beetle, Tribolium castaneum.

    PubMed

    Via, S

    1999-03-01

    Cannibalism is well known to affect both the population dynamics and the competitive relationships of organisms. Cannibalistic behaviour commonly increases in stressful conditions, such as when density is high or food is scarce, and cannibals often obtain a nutritional benefit. Might cannibalism also increase in a novel environment to which a population is poorly adapted physiologically? Moreover, might cannibalistic behaviour provide enough of a nutritional advantage in a nutritionally stressful environment to rescue individuals from its adverse effects and thus permit colonization and range expansion? Previous work has shown that oat flour is a particularly stressful environment for Tribolium castaneum. In the study reported here, egg cannibalism by two strains of T. castaneum was significantly enhanced in oat flour, and egg eating rescued larvae from the adverse demographic effects of this poor environment. Development time of the cannibals was accelerated almost to the level seen for individuals reared in the nutritionally superior environment (wheat plus brewer's yeast). Their survival and fecundity also increased relative to individuals reared in oat flour without the opportunity to cannibalize. A sib analysis revealed that for larvae reared in the presence of victim eggs, the extent of cannibalism was genetically variable, so that this trait could evolve, given a selective benefit exceeding its cost. These results suggest that colonization of a marginal new environment could be facilitated by enhanced rates of cannibalism. The possible interplay between cannibalism and physiological adaptation to a new environment is discussed.

  13. Signaling Properties and Pharmacological Analysis of Two Sulfakinin Receptors from the Red Flour Beetle, Tribolium castaneum

    PubMed Central

    Zels, Sven; Verlinden, Heleen; Dillen, Senne; Vleugels, Rut; Nachman, Ronald J.; Broeck, Jozef Vanden

    2014-01-01

    Sulfakinin is an insect neuropeptide that constitutes an important component of the complex network of hormonal and neural factors that regulate feeding and digestion. The key modulating functions of sulfakinin are mediated by binding and signaling via G-protein coupled receptors. Although a substantial amount of functional data have already been reported on sulfakinins in different insect species, only little information is known regarding the properties of their respective receptors. In this study, we report on the molecular cloning, functional expression and characterization of two sulfakinin receptors in the red flour beetle, Tribolium castaneum. Both receptor open reading frames show extensive sequence similarity with annotated sulfakinin receptors from other insects. Comparison of the sulfakinin receptor sequences with homologous vertebrate cholecystokinin receptors reveals crucial conserved regions for ligand binding and receptor activation. Quantitative reverse transcriptase PCR shows that transcripts of both receptors are primarily expressed in the central nervous system of the beetle. Pharmacological characterization using 29 different peptide ligands clarified the essential requirements for efficient activation of these sulfakinin receptors. Analysis of the signaling pathway in multiple cell lines disclosed that the sulfakinin receptors of T. castaneum can stimulate both the Ca2+ and cyclic AMP second messenger pathways. This in depth characterization of two insect sulfakinin receptors may provide useful leads for the further development of receptor ligands with a potential applicability in pest control and crop protection. PMID:24718573

  14. Identification, cloning, and expression of a GHF9 cellulase from Tribolium castaneum (Coleoptera: Tenebrionidae).

    PubMed

    Willis, Jonathan D; Oppert, Brenda; Oppert, Cris; Klingeman, William E; Jurat-Fuentes, Juan L

    2011-02-01

    The availability of sequenced insect genomes has allowed for discovery and functional characterization of novel genes and proteins. We report use of the Tribolium castaneum (Herbst) (red flour beetle) genome to identify, clone, express, and characterize a novel endo-β-1,4-glucanase we named TcEG1 (T. castaneum endoglucanase 1). Sequence analysis of a full-length TcEG1 cDNA clone (1356bp) revealed sequence homology to enzymes in glycosyl hydrolase family 9 (GHF9), and verified presence of a change (Gly for Ser) in the conserved catalytic domain for GHF9 cellulases. This TcEG1 cDNA clone was predicted to encode a 49.5kDa protein with a calculated pI of 5.39. Heterologous expression of TcEG1 in Drosophila S2 cell cultures resulted in secretion of a 51-kDa protein, as determined by Western blotting. The expressed protein was used to characterize TcEG1 enzymatic activity against two cellulose substrates to determine its specificity and stability. Our data support that TcEG1 as a novel endo-β-1,4-glucanase, the first functional characterization of a cellulase enzyme derived from an insect genome with potential applications in the biofuel industry due to its high relative activity at alkaline pH.

  15. Activity of Schinus areira (Anacardiaceae) essential oils against the grain storage pest Tribolium castaneum.

    PubMed

    Descamps, Lilian R; Sánchez Chopa, Carolina; Ferrero, Adriana A

    2011-06-01

    Essential oils extracted from leaves and fruits of Schinus areira (Anacardiaceae) were tested for their repellent, toxic and feeding deterrent properties against Tribolium castaneum (Coleoptera: Tenebrionidae) larvae and adults. A topical application assay was employed for the contact toxicity study and filter paper impregnation for the fumigant assay. A treated diet was also used to evaluate the repellent activity and a flour disk bioassay for the feeding deterrent action and nutritional index alteration. The essential oil of the leaves contained mainly monoterpenoids, with alpha-phellandrene, 3-carene and camphene predominant, whereas that from the fruits contained mainly alpha-phellandrene, 3-carene and beta-myrcene. The leaf essential oil showed repellent effects, whereas that from the fruit was an attractant. Both oils produced mortality against larvae in topical and fumigant bioassays, but fumigant toxicity was not found against adults. Moreover, both essential oils produced some alterations in nutritional index. These results show that the essential oils from S. areira could be applicable to the management of populations of Tribolium castaneum.

  16. Efficacy of aerosol applications of methoprene and synergized pyrethrin against Tribolium castaneum adults and eggs.

    PubMed

    Tucker, Angela M; Campbell, James F; Arthur, Frank H; Zhu, Kun Yan

    2014-06-01

    Experiments were performed to determine the efficacy of a single aerosol application of the insecticides methoprene and piperonyl butoxide-synergized pyrethrin, alone or in combination, and the insecticide carrier, Isopar M, against Tribolium castaneum (Herbst), the red flour beetle. The initial test exposed adults to insecticide treatments and placed male/female pairs in flour. All adults exposed to synergized pyrethrin were knocked down for at least 24 h after exposure but they recovered. High adult survival and similar average numbers of living F1 progeny were produced regardless of treatment exposure. In a separate test, insecticide treatments were directly applied to newly laid eggs, which resulted in the suppression of egg hatch. Synergized pyrethrin was the most effective insecticide (P < or = 0.001) for suppressing egg hatch. The effect of flour on insecticide activity to eggs and consequent insect development was also evaluated. An amount of 0.01 g of flour in the exposure arena, 62-cm2 area, was not sufficient for individuals to develop beyond the early larval stages, regardless of the treatment. As the flour amount in the arena increased from 1 to 5 g, the number of eggs that could develop to the adult stage increased, but this number was significantly lower in the insecticide treatments than in the control or carrier treatments. The results of the later tests indicate a high efficacy of the insecticides alone or in combination on T. castaneum egg hatch and development to the adult stage.

  17. Pharmacological and signalling properties of a D2-like dopamine receptor (Dop3) in Tribolium castaneum.

    PubMed

    Verlinden, Heleen; Vleugels, Rut; Verdonck, Rik; Urlacher, Elodie; Vanden Broeck, Jozef; Mercer, Alison

    2015-01-01

    Dopamine is an important neurotransmitter in the central nervous system of vertebrates and invertebrates. Despite their evolutionary distance, striking parallels exist between deuterostomian and protostomian dopaminergic systems. In both, signalling is achieved via a complement of functionally distinct dopamine receptors. In this study, we investigated the sequence, pharmacology and tissue distribution of a D2-like dopamine receptor from the red flour beetle Tribolium castaneum (TricaDop3) and compared it with related G protein-coupled receptors in other invertebrate species. The TricaDop3 receptor-encoding cDNA shows considerable sequence similarity with members of the Dop3 receptor class. Real time qRT-PCR showed high expression in both the central brain and the optic lobes, consistent with the role of dopamine as neurotransmitter. Activation of TricaDop3 expressed in mammalian cells increased intracellular Ca(2+) signalling and decreased NKH-477 (a forskolin analogue)-stimulated cyclic AMP levels in a dose-dependent manner. We studied the pharmacological profile of the TricaDop3 receptor and demonstrated that the synthetic vertebrate dopamine receptor agonists, 2 - amino- 6,7 - dihydroxy - 1,2,3,4 - tetrahydronaphthalene hydrobromide (6,7-ADTN) and bromocriptine acted as agonists. Methysergide was the most potent of the antagonists tested and showed competitive inhibition in the presence of dopamine. This study offers important information on the Dop3 receptor from Tribolium castaneum that will facilitate functional analyses of dopamine receptors in insects and other invertebrates.

  18. Identification and evolution of two insulin receptor genes involved in Tribolium castaneum development and reproduction.

    PubMed

    Sang, Ming; Li, Chengjun; Wu, Wei; Li, Bin

    2016-07-10

    The insulin and insulin-like signaling (IIS) pathway exists in a wide range of organisms from mammals to invertebrates and regulates several vital physiological functions. A phylogenetic analysis have indicated that insulin receptors have been duplicated at least twice among vertebrates, whereas only one duplication occurred in insects before the differentiation of Coleoptera, Hymenoptera, and Hemiptera. Thus, we cloned two putative insulin receptor genes, T.cas-ir1 and T.cas-ir2, from T. castaneum and determined that T.cas-ir1 is most strongly expressed during the late adult and early pupal stages, whereas T.cas-ir2 is most strongly expressed during the late larval stage. We found that larval RNAi against T.cas-ir1 and T.cas-ir2 causes 100% and 42.0% insect death, respectively, and that parental RNAi against T.cas-ir1 and T.cas-ir2 leads to 100% and 33.3% reductions in beetle fecundity, respectively. The hatching rate of ds-ir2 insects was 66.2%. Moreover, RNAi against these two genes increased the expression of the pkc, foxo, jnk, cdc42, ikk, and mekk genes but decreased erk gene expression. Despite these similarities, these two genes act via distinct regulatory pathways. These results indicate that these two receptors have functionally diverged with respect to the development and reproduction of T. castaneum, even though they retain some common regulatory signaling pathways.

  19. Comparative RNA-sequencing analysis of mthl1 functions and signal transductions in Tribolium castaneum.

    PubMed

    Li, Chengjun; Wu, Wei; Sang, Ming; Liu, Xing; Hu, Xingxing; Yun, Xiaopei; Li, Bin

    2014-09-01

    Methuselah-like 1 of Tribolium castaneum (TcMthl1) has been reported to play crucial roles in development, lifespan, stress resistance and reproduction. However, the signaling system of TcMthl1 is unknown. Thus, we compare the transcriptome profile of RNAi treated larvae (ds-Tcmthl1) and control larvae of T. castaneum by RNA-sequencing, and obtained 14,613,514 sequence reads aligned with 13,533 genes; 812 differentially expressed genes (DEGs) were identified. These DEGs were classified into 47 GO functional groups, including such functions as the immune system process, the response to stimulus, the developmental process and reproduction. Interestingly, knock-down of Tcmthl1 suppressed both of Toll and IMD immunity pathways which most likely modulated the effects of Tcmthl1 on lifespan and stress resistance. Additionally, the DEGs encoding Blimp-1, Ftz-F1, E74 and Timeless may participate in the development and reproduction of ds-Tcmthl1 insects. The findings of these DEGs and pathways will provide valuable insight into TcMthl1 signaling and regulation system.

  20. The stimulation of immune defence accelerates development in the red flour beetle (Tribolium castaneum).

    PubMed

    Roth, O; Kurtz, J

    2008-11-01

    The timing of the transition between life stages is of key importance for an organism. Depending on the environmental conditions, maturing earlier at a smaller size or maturing later at a larger size can be advantageous for fitness. Exposure to parasites and subsequent immune activation may lead to alterations in development. Immune defence often comes at a cost, such as energy drain towards immune function, which is likely to delay development. On the other hand, animals may react to an anticipated risk of infection with a phenotypically plastic shift in life history, which may more likely lead to accelerated development and earlier maturation. We tested these alternatives in the red flour beetle, Tribolium castaneum. Young larvae were exposed to a non-infectious immune challenge with heat-killed bacteria (either Escherichia coli or Bacillus thuringiensis) and they were followed up for their development, survival, adult size and reproduction. We found that animals that had experienced a bacterial challenge developed into adults earlier than sham-treated beetles, while they did not differ significantly in survival or adult size. Beetles exposed to E. coli produced fewer offspring, while exposure to B. thuringiensis did not affect offspring number. Taken together, our results indicate that T. castaneum is able to speed up its development when facing a risk of infection.

  1. Experimental evolution exposes female and male responses to sexual selection and conflict in Tribolium castaneum.

    PubMed

    Michalczyk, Łukasz; Millard, Anna L; Martin, Oliver Y; Lumley, Alyson J; Emerson, Brent C; Gage, Matthew J G

    2011-03-01

    Between-individual variance in potential reproductive rate theoretically creates a load in reproducing populations by driving sexual selection of male traits for winning competitions, and female traits for resisting the costs of multiple mating. Here, using replicated experimental evolution under divergent operational sex ratios (OSR, 9:1 or 1:6 ♀:♂) we empirically identified the parallel reproductive fitness consequences for females and males in the promiscuous flour beetle Tribolium castaneum. Our results revealed clear evidence that sexual conflict resides within the T. castaneum mating system. After 20 generations of selection, females from female-biased OSRs became vulnerable to multiple mating, and showed a steep decrease in reproductive fitness with an increasing number of control males. In contrast, females from male-biased OSRs showed no change in reproductive fitness, irrespective of male numbers. The divergence in reproductive output was not explained by variation in female mortality. Parallel assays revealed that males also responded to experimental evolution: individuals from male-biased OSRs obtained 27% greater reproductive success across 7-day competition for females with a control male rival, compared to males from the female-biased lines. Subsequent assays suggest that these differences were not due to postcopulatory sperm competitiveness, but to precopulatory/copulatory competitive male mating behavior. PMID:21091981

  2. Repellent activity of essential oils and some of their individual constituents against Tribolium castaneum herbst.

    PubMed

    Caballero-Gallardo, Karina; Olivero-Verbel, Jesús; Stashenko, Elena E

    2011-03-01

    A tool for integrated pest management is the use of essential oils (EOs) and plant extracts. In this study, EOs from Tagetes lucida , Lepechinia betonicifolia , Lippia alba , Cananga odorata , and Rosmarinus officinalis , species grown in Colombia, were analyzed by gas chromatography-mass spectrometry. These oils as well as several of their constituents were tested for repellent activity against Tribolium castaneum , using the area preference method. The main components (>10%) found in EOs were methylchavicol, limonene/α-pinene, carvone/limonene, benzyl acetate/linalool/benzyl benzoate, and α-pinene, for T. lucida, L. betonicifolia, L. alba, C. odorata, and R. officinalis, respectively. All EOs were repellent, followed a dose-response relationship, and had bioactivity similar to or better than that of commercial compound IR3535. EOs from C. odorata and L. alba were the most active. Compounds from EOs, such benzyl benzoate, β-myrcene, and carvone, showed good repellent properties. In short, EOs from plants cultivated in Colombia are sources of repellents against T. castaneum. PMID:21291237

  3. Insecticidal properties of essential oils against Tribolium castaneum (Herbst) and their inhibitory effects on acetylcholinesterase and adenosine triphosphatases.

    PubMed

    Abou-Taleb, Hamdy K; Mohamed, Magdy I E; Shawir, Mohamed S; Abdelgaleil, Samir A M

    2016-01-01

    Essential oils from 20 Egyptian plants were obtained by using hydrodistillation. The chemical composition of the isolated oils was identified by gas chromatograph/mass spectrometer. Fumigant and contact toxicities of the essential oils were evaluated against the adults of Tribolium castaneum. In fumigation assays, the oil of Origanum vulgare (LC50 = 9.97 mg/L air) displayed the highest toxicity towards the adults of T. castaneum. In contact assays, the oils of Artemisia monosperma (LC50 = 0.07 mg/cm(2)) and O. vulgare (LC50 = 0.07 mg/cm(2)) were the most potent toxicants against the adults of T. castaneum. Biochemical studies showed that the tested oils caused pronounced inhibition of acetylcholinesterase (AChE) and adenosine triphosphatases (ATPases) isolated from the larvae of T. castaneum. The oil Cupressus macrocarpa (IC50 = 12.3 mg/L) was the most potent inhibitor of AChE, while the oil of Calistemon viminals (IC50 = 4.4 mg/L) was the most potent inhibitor of ATPases. PMID:25978134

  4. Functions of Ion Transport Peptide and Ion Transport Peptide-Like in the Red Flour Beetle Tribolium castaneum

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Ion transport peptide (ITP) and ITP-like (ITPL) are highly conserved neuropeptides in insects and crustaceans. We investigated the alternatively spliced variants of ITP/ITPL in Tribolium castaneum to understand their functions. We identified three alternatively spliced transcripts named itp, itpl-...

  5. Superoxide dismutase 2 knockdown leads to defects in locomotor activity, sensitivity to paraquat, and increased cuticle pigmentation in Tribolium castaneum.

    PubMed

    Tabunoki, Hiroko; Gorman, Maureen J; Dittmer, Neal T; Kanost, Michael R

    2016-01-01

    Insects can rapidly adapt to environmental changes through physiological responses. The red flour beetle Tribolium castaneum is widely used as a model insect species. However, the stress-response system of this species remains unclear. Superoxide dismutase 2 (SOD2) is a crucial antioxidative enzyme that is found in mitochondria. T. castaneum SOD2 (TcSOD2) is composed of 215 amino acids, and has an iron/manganese superoxide dismutase domain. qRT-PCR experiments revealed that TcSOD2 was present through all developmental stages. To evaluate TcSOD2 function in T. castaneum, we performed RNAi and also assessed the phenotype and antioxidative tolerance of the knockdown of TcSOD2 by exposing larvae to paraquat. The administration of paraquat resulted in significantly higher 24-h mortality in TcSOD2 knockdown larval groups than in the control groups. The TcSOD2 knockdown adults moved significantly more slowly, had lower ATP content, and exhibited a different body color from the control groups. We found that TcSOD2 dsRNA treatment in larvae resulted in increased expression of tyrosinase and laccase2 mRNA after 10 days. This is the first report showing that TcSOD2 has an antioxidative function and demonstrates that T. castaneum may use an alternative antioxidative system when the SOD2-based system fails. PMID:27387523

  6. Insecticidal properties of essential oils against Tribolium castaneum (Herbst) and their inhibitory effects on acetylcholinesterase and adenosine triphosphatases.

    PubMed

    Abou-Taleb, Hamdy K; Mohamed, Magdy I E; Shawir, Mohamed S; Abdelgaleil, Samir A M

    2016-01-01

    Essential oils from 20 Egyptian plants were obtained by using hydrodistillation. The chemical composition of the isolated oils was identified by gas chromatograph/mass spectrometer. Fumigant and contact toxicities of the essential oils were evaluated against the adults of Tribolium castaneum. In fumigation assays, the oil of Origanum vulgare (LC50 = 9.97 mg/L air) displayed the highest toxicity towards the adults of T. castaneum. In contact assays, the oils of Artemisia monosperma (LC50 = 0.07 mg/cm(2)) and O. vulgare (LC50 = 0.07 mg/cm(2)) were the most potent toxicants against the adults of T. castaneum. Biochemical studies showed that the tested oils caused pronounced inhibition of acetylcholinesterase (AChE) and adenosine triphosphatases (ATPases) isolated from the larvae of T. castaneum. The oil Cupressus macrocarpa (IC50 = 12.3 mg/L) was the most potent inhibitor of AChE, while the oil of Calistemon viminals (IC50 = 4.4 mg/L) was the most potent inhibitor of ATPases.

  7. Identification and characterization of novel ER-based hsp90 gene in the red flour beetle, Tribolium castaneum.

    PubMed

    Zhang, Yi; Gu, Shasha; Li, Chengjun; Sang, Ming; Wu, Wei; Yun, Xiaopei; Hu, Xingxing; Li, Bin

    2014-09-01

    Heat-shock protein 90 (HSP90) is a highly conserved molecular chaperone found in all species except for Archaea, which is required not only for stress tolerance but also for normal development. Recently, it was reported that HSP83, one member of the cytosolic HSP90 family, contributes to oogenesis and responds to heat resistance in Tribolium castaneum. Here, a novel ER-based HSP90 gene, Tchsp90, has been identified in T. castaneum. Phylogenetic analysis showed that hsp90s and hsp83s evolved separately from a common ancestor but that hsp90s originated earlier. Quantitative real-time polymerase chain reaction illustrated that Tchsp90 is expressed in all developmental stages and is highly expressed at early pupa and late adult stages. Tchsp90 was upregulated in response to heat stress but not to cold stress. Laval RNAi revealed that Tchsp90 is important for larval/pupal development. Meanwhile, parental RNAi indicated that it completely inhibited female fecundity and partially inhibited male fertility once Tchsp90 was knocked down and that it will further shorten the lifespan of T. castaneum. These results suggest that Tchsp90 is essential for development, lifespan, and reproduction in T. castaneum in addition to its response to heat stress.

  8. Superoxide dismutase 2 knockdown leads to defects in locomotor activity, sensitivity to paraquat, and increased cuticle pigmentation in Tribolium castaneum

    PubMed Central

    Tabunoki, Hiroko; Gorman, Maureen J.; Dittmer, Neal T.; Kanost, Michael R.

    2016-01-01

    Insects can rapidly adapt to environmental changes through physiological responses. The red flour beetle Tribolium castaneum is widely used as a model insect species. However, the stress–response system of this species remains unclear. Superoxide dismutase 2 (SOD2) is a crucial antioxidative enzyme that is found in mitochondria. T. castaneum SOD2 (TcSOD2) is composed of 215 amino acids, and has an iron/manganese superoxide dismutase domain. qRT-PCR experiments revealed that TcSOD2 was present through all developmental stages. To evaluate TcSOD2 function in T. castaneum, we performed RNAi and also assessed the phenotype and antioxidative tolerance of the knockdown of TcSOD2 by exposing larvae to paraquat. The administration of paraquat resulted in significantly higher 24-h mortality in TcSOD2 knockdown larval groups than in the control groups. The TcSOD2 knockdown adults moved significantly more slowly, had lower ATP content, and exhibited a different body color from the control groups. We found that TcSOD2 dsRNA treatment in larvae resulted in increased expression of tyrosinase and laccase2 mRNA after 10 days. This is the first report showing that TcSOD2 has an antioxidative function and demonstrates that T. castaneum may use an alternative antioxidative system when the SOD2-based system fails. PMID:27387523

  9. Food source provisioning and susceptibility of immature and adult Tribolium castaneum on concrete partially treated with chlorfenapyr (Phantom®)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A series of experiments were conducted in which adults, pupae, and 4-week-old larvae of Tribolium castaneum (Herbst), the red flour beetle, were exposed separately on concrete arenas partially treated (14.4 % of the total area) with the insecticide chlorfenapyr (Phantom®) at 1.1 g active ingredient/...

  10. Effects of flour and milling debris on efficacy of beta-cyfluthrin for control of Tribolium castaneum

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Laboratory and field studies were conducted to determine if accumulation of a flour food source or milling debris affected residual efficacy of beta-cyfluthrin for control of Tribolium castaneum (Herbst), the red flour beetle. In the laboratory study, the high label rate of 20 mg active ingredient (...

  11. Contact toxicity of deltamethrin against Tribolium castaneum (Coleoptera: Tenebrionidae), Sitophilus oryzae (Coleoptera: Curculionidae), and Rhyzopertha dominica (Coleoptera: Bostrichidae) adults

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This study was conducted to evaluate deltamethrin-incorporated ZeroFly® Storage Bags for efficacy against stored-product insect pests. We evaluated response to deltamethrin concentrations for adults of three stored-product insects, Tribolium castaneum (Herbst), Sitophilus oryzae (L.), and Rhyzoperth...

  12. Capture of Tribolium castaneum and Tribolium confusum (Coleoptera: Tenebrionidae) in floor traps: the effect of previous captures

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The impact of prior captures on the trapping performance of floor traps was evaluated for the red flour beetle, Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae) and the confused flour beetle, Tribolium confusum Jacquelin du Val (Coleoptera: Tenebrionidae) in laboratory conditions. The effect...

  13. The presence of flour affects the efficacy of aerosolized insecticides used to treat the red flour beetle, Tribolium castaneum

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Experiments were conducted in tightly sealed pilot-scale warehouses to assess the efficacy of common aerosolized insecticides on all life stages of Tribolium castaneum when exposed in dishes containing 0 to 2 g of wheat flour either under pallets or out in the open. Petri dishes containing 0, 0.1, ...

  14. Live imaging of Tribolium castaneum embryonic development using light-sheet-based fluorescence microscopy.

    PubMed

    Strobl, Frederic; Schmitz, Alexander; Stelzer, Ernst H K

    2015-10-01

    Tribolium castaneum has become an important insect model organism for evolutionary developmental biology, genetics and biotechnology. However, few protocols for live fluorescence imaging of Tribolium have been reported, and little image data is available. Here we provide a protocol for recording the development of Tribolium embryos with light-sheet-based fluorescence microscopy. The protocol can be completed in 4-7 d and provides procedural details for: embryo collection, microscope configuration, embryo preparation and mounting, noninvasive live imaging for up to 120 h along multiple directions, retrieval of the live embryo once imaging is completed, and image data processing, for which exemplary data is provided. Stringent quality control criteria for developmental biology studies are also discussed. Light-sheet-based fluorescence microscopy complements existing toolkits used to study Tribolium development, can be adapted to other insect species, and requires no advanced imaging or sample preparation skills.

  15. A system to efficiently maintain embryonic lethal mutations in the flour beetle Tribolium castaneum.

    PubMed

    Berghammer, A; Bucher, G; Maderspacher, F; Klingler, M

    1999-06-01

    Due to its small size, short life cycle, and easy maintenance, the flour beetle Tribolium castaneum is well suited for the genetic analysis of development. One drawback of Tribolium as a genetic system is, however, the difficulty of keeping embryonic lethal lines. Presently, only few lethal mutations can be kept as balanced stocks. Therefore, heterozygous carriers must be identified anew in every generation in order to maintain a recessive embryonic mutation. To alleviate this problem we have devised a block system that allows the simultaneous processing of many mutant lines or test crosses for visual inspection of larval cuticle phenotypes. Using this technique, one person can maintain about 100 embryonic lethal stocks, which makes feasible the thorough genetic analysis of embryogenesis in this species.

  16. Genome-wide mapping of conserved microRNAs and their host transcripts in Tribolium castaneum.

    PubMed

    Luo, Qibin; Zhou, Qing; Yu, Xiaomin; Lin, Hongbin; Hu, Songnian; Yu, Jun

    2008-06-01

    MicroRNAs (miRNAs) are endogenous 22-nt RNAs, which play important regulatory roles by post-transcriptional gene silencing. A computational strategy has been developed for the identification of conserved miRNAs based on features of known metazoan miRNAs in red flour beetle (Tribolium castaneum), which is regarded as one of the major laboratory models of arthropods. Among 118 putative miRNAs, 47% and 53% of the predicted miRNAs from the red flour beetle are harbored by known protein-coding genes (intronic) and genes located outside (intergenic miRNA), respectively. There are 31 intronic miRNAs in the same transcriptional orientation as the host genes, which may share RNA polymerase II and spliceosomal machinery with their host genes for their biogenesis. A hypothetical feedback model has been proposed based on the analysis of the relationship between intronic miRNAs and their host genes in the development of red flour beetle.

  17. Satellite DNA Modulates Gene Expression in the Beetle Tribolium castaneum after Heat Stress.

    PubMed

    Feliciello, Isidoro; Akrap, Ivana; Ugarković, Đurđica

    2015-08-01

    Non-coding repetitive DNAs have been proposed to perform a gene regulatory role, however for tandemly repeated satellite DNA no such role was defined until now. Here we provide the first evidence for a role of satellite DNA in the modulation of gene expression under specific environmental conditions. The major satellite DNA TCAST1 in the beetle Tribolium castaneum is preferentially located within pericentromeric heterochromatin but is also dispersed as single repeats or short arrays in the vicinity of protein-coding genes within euchromatin. Our results show enhanced suppression of activity of TCAST1-associated genes and slower recovery of their activity after long-term heat stress relative to the same genes without associated TCAST1 satellite DNA elements. The level of gene suppression is not influenced by the distance of TCAST1 elements from the associated genes up to 40 kb from the genes' transcription start sites, but it does depend on the copy number of TCAST1 repeats within an element, being stronger for the higher number of copies. The enhanced gene suppression correlates with the enrichment of the repressive histone marks H3K9me2/3 at dispersed TCAST1 elements and their flanking regions as well as with increased expression of TCAST1 satellite DNA. The results reveal transient, RNAi based heterochromatin formation at dispersed TCAST1 repeats and their proximal regions as a mechanism responsible for enhanced silencing of TCAST1-associated genes. Differences in the pattern of distribution of TCAST1 elements contribute to gene expression diversity among T. castaneum strains after long-term heat stress and might have an impact on adaptation to different environmental conditions. PMID:26275223

  18. Tribolium castaneum Transformer-2 regulates sex determination and development in both males and females.

    PubMed

    Shukla, Jayendra Nath; Palli, Subba Reddy

    2013-12-01

    Tribolium castaneum Transformer (TcTra) is essential for female sex determination and maintenance through the regulation of sex-specific splicing of doublesex (dsx) pre-mRNA. In females, TcTra also regulates the sex-specific splicing of its own pre-mRNA to ensure continuous production of functional Tra protein. Transformer protein is absent in males and hence dsx pre-mRNA is spliced in a default mode. The mechanisms by which males inhibit the production of functional Tra protein are not known. Here, we report on functional characterization of transformer-2 (tra-2) gene (an ortholog of Drosophila transformer-2) in T. castaneum. RNA interference-mediated knockdown in the expression of gene coding for tra-2 in female pupae or adults resulted in the production of male-specific isoform of dsx and both female and male isoforms of tra suggesting that Tra-2 is essential for the female-specific splicing of tra and dsx pre-mRNAs. Interestingly, knockdown of tra-2 in males did not affect the splicing of dsx but resulted in the production of both female and male isoforms of tra suggesting that Tra-2 suppresses female-specific splicing of tra pre-mRNA in males. This dual regulation of sex-specific splicing of tra pre-mRNA ensures a tight regulation of sex determination and maintenance. These data suggest a critical role for Tra-2 in suppression of female sex determination cascade in males. In addition, RNAi studies showed that Tra-2 is also required for successful embryonic and larval development in both sexes.

  19. Satellite DNA-like elements associated with genes within euchromatin of the beetle Tribolium castaneum.

    PubMed

    Brajković, Josip; Feliciello, Isidoro; Bruvo-Mađarić, Branka; Ugarković, Durđica

    2012-08-01

    In the red flour beetle Tribolium castaneum the major TCAST satellite DNA accounts for 35% of the genome and encompasses the pericentromeric regions of all chromosomes. Because of the presence of transcriptional regulatory elements and transcriptional activity in these sequences, TCAST satellite DNAs also have been proposed to be modulators of gene expression within euchromatin. Here, we analyze the distribution of TCAST homologous repeats in T. castaneum euchromatin and study their association with genes as well as their potential gene regulatory role. We identified 68 arrays composed of TCAST-like elements distributed on all chromosomes. Based on sequence characteristics the arrays were composed of two types of TCAST-like elements. The first type consists of TCAST satellite-like elements in the form of partial monomers or tandemly arranged monomers, up to tetramers, whereas the second type consists of TCAST-like elements embedded with a complex unit that resembles a DNA transposon. TCAST-like elements were also found in the 5' untranslated region (UTR) of the CR1-3_TCa retrotransposon, and therefore retrotransposition may have contributed to their dispersion throughout the genome. No significant difference in the homogenization of dispersed TCAST-like elements was found either at the level of local arrays or chromosomes nor among different chromosomes. Of 68 TCAST-like elements, 29 were located within introns, with the remaining elements flanked by genes within a 262 to 404,270 nt range. TCAST-like elements are statistically overrepresented near genes with immunoglobulin-like domains attesting to their nonrandom distribution and a possible gene regulatory role. PMID:22908042

  20. Satellite DNA-Like Elements Associated With Genes Within Euchromatin of the Beetle Tribolium castaneum

    PubMed Central

    Brajković, Josip; Feliciello, Isidoro; Bruvo-Mađarić, Branka; Ugarković, Đurđica

    2012-01-01

    In the red flour beetle Tribolium castaneum the major TCAST satellite DNA accounts for 35% of the genome and encompasses the pericentromeric regions of all chromosomes. Because of the presence of transcriptional regulatory elements and transcriptional activity in these sequences, TCAST satellite DNAs also have been proposed to be modulators of gene expression within euchromatin. Here, we analyze the distribution of TCAST homologous repeats in T. castaneum euchromatin and study their association with genes as well as their potential gene regulatory role. We identified 68 arrays composed of TCAST-like elements distributed on all chromosomes. Based on sequence characteristics the arrays were composed of two types of TCAST-like elements. The first type consists of TCAST satellite-like elements in the form of partial monomers or tandemly arranged monomers, up to tetramers, whereas the second type consists of TCAST-like elements embedded with a complex unit that resembles a DNA transposon. TCAST-like elements were also found in the 5′ untranslated region (UTR) of the CR1-3_TCa retrotransposon, and therefore retrotransposition may have contributed to their dispersion throughout the genome. No significant difference in the homogenization of dispersed TCAST-like elements was found either at the level of local arrays or chromosomes nor among different chromosomes. Of 68 TCAST-like elements, 29 were located within introns, with the remaining elements flanked by genes within a 262 to 404,270 nt range. TCAST-like elements are statistically overrepresented near genes with immunoglobulin-like domains attesting to their nonrandom distribution and a possible gene regulatory role. PMID:22908042

  1. Susceptibility of Tribolium castaneum (Coleoptera: Tenebrionidae) life stages to flameless catalytic infrared radiation.

    PubMed

    Khamis, Moses; Subramanyam, Bhadriraju; Flinn, Paul W; Dogan, Hulya; Gwirtz, Jeffrey A

    2011-02-01

    The susceptibility of various life stages of the red flour beetle, Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae), a pest of stored wheat, Triticum aestivum L., to flameless catalytic infrared radiation in the 3-7-microm range was evaluated in the laboratory. Immature stages were collected from flour infested with T. castaneum adults only for 1 d. Stages collected after 1 d represented eggs (collected on day 0); those collected after 7, 14, and 21 d from day 0 represented larvae in different developmental stages, whereas those collected after 24 d represented pupae. Adults (2 wk old) were collected after 42 d. Each of these stages was exposed for 45 or 60 s in 113.5 or 227.0 g of wheat at a distance of 8.0 or 12.7 cm from a bench top infrared emitter. The mean temperatures attained during exposures were measured continuously using a noncontact infrared thermometer connected to a computer. The mean grain temperatures attained increased with an increase in exposure time and were inversely related to distance from the emitter. Grain quantity least influenced mean temperatures attained. Pupae were the least susceptible stage and larvae collected after 7 d were the most susceptible stage. Variation in probability of death of various life stages decreased with an increase in mean grain temperatures attained. All life stages were killed after a 60-s exposure at a distance of 8.0 cm from the emitter in 113.5 g of wheat, where the mean +/- SE temperatures attained ranged from 107.6 +/- 1.2 to 111.4 +/- 0.5 degrees C. Our laboratory results using small grain quantities and short exposure times showed that flameless catalytic infrared radiation can be a valuable tool for managing insects in stored organic and nonorganic wheat.

  2. Response of body size and developmental time of Tribolium castaneum to constant versus fluctuating thermal conditions.

    PubMed

    Małek, D; Drobniak, S; Gozdek, A; Pawlik, K; Kramarz, P

    2015-07-01

    Temperature has profound effects on biological functions at all levels of organization. In ectotherms, body size is usually negatively correlated with ambient temperature during development, a phenomenon known as The Temperature-Size Rule (TSR). However, a growing number of studies have indicated that temperature fluctuations have a large influence on life history traits and the implications of such fluctuations for the TSR are unknown. Our study investigated the effect of different constant and fluctuating temperatures on the body mass and development time of red flour beetles (Tribolium castaneum Herbst, 1797); we also examined whether the sexes differed in their responses to thermal conditions. We exposed the progeny of half-sib families of a T. castaneum laboratory strain to one of four temperature regimes: constant 30°C, constant 25°C, fluctuating with a daily mean of 30°C, or fluctuating with a daily mean of 25°C. Sex-specific development time and body mass at emergence were determined. Beetles developed the fastest and had the greatest body mass upon emergence when they were exposed to a constant temperature of 30°C. This pattern was reversed when beetles experienced a constant temperature of 25°C: slowest development and lowest body mass upon emergence were observed. Fluctuations changed those effects significantly - impact of temperature on development time was smaller, while differences in body mass disappeared completely. Our results do not fit TSR predictions. Furthermore, regardless of the temperature regime, females acquired more mass, while there were no differences between sexes in development time to eclosion. This finding fails to support one of the explanations for smaller male size: that selection favors the early emergence of males. We found no evidence of genotype × environment interactions for selected set of traits.

  3. Satellite DNA Modulates Gene Expression in the Beetle Tribolium castaneum after Heat Stress.

    PubMed

    Feliciello, Isidoro; Akrap, Ivana; Ugarković, Đurđica

    2015-08-01

    Non-coding repetitive DNAs have been proposed to perform a gene regulatory role, however for tandemly repeated satellite DNA no such role was defined until now. Here we provide the first evidence for a role of satellite DNA in the modulation of gene expression under specific environmental conditions. The major satellite DNA TCAST1 in the beetle Tribolium castaneum is preferentially located within pericentromeric heterochromatin but is also dispersed as single repeats or short arrays in the vicinity of protein-coding genes within euchromatin. Our results show enhanced suppression of activity of TCAST1-associated genes and slower recovery of their activity after long-term heat stress relative to the same genes without associated TCAST1 satellite DNA elements. The level of gene suppression is not influenced by the distance of TCAST1 elements from the associated genes up to 40 kb from the genes' transcription start sites, but it does depend on the copy number of TCAST1 repeats within an element, being stronger for the higher number of copies. The enhanced gene suppression correlates with the enrichment of the repressive histone marks H3K9me2/3 at dispersed TCAST1 elements and their flanking regions as well as with increased expression of TCAST1 satellite DNA. The results reveal transient, RNAi based heterochromatin formation at dispersed TCAST1 repeats and their proximal regions as a mechanism responsible for enhanced silencing of TCAST1-associated genes. Differences in the pattern of distribution of TCAST1 elements contribute to gene expression diversity among T. castaneum strains after long-term heat stress and might have an impact on adaptation to different environmental conditions.

  4. Fine-scale analysis of parasite resistance genes in the red flour beetle, Tribolium castaneum.

    PubMed

    Zhong, Daibin; Pai, Aditi; Wang, Mei-Hui; Keech, Naomi; Yan, Guiyun

    2013-09-01

    Parasite infection impacts population dynamics through effects on fitness and fecundity of the individual host. In addition to the known roles of environmental factors, host susceptibility to parasites has a genetic basis that has not been well characterized. We previously mapped quantitative trait loci (QTL) for susceptibility to rat tapeworm (Hymenolepis diminuta) infection in Tribolium castaneum using dominant AFLP markers; however, the resistance genes were not identified. Here, we refined the QTL locations and increased the marker density in the QTL regions using new microsatellite markers, sequence-tagged site markers, and single-strand conformational polymorphism markers. Resistance QTL in three linkage groups (LG3, LG6, and LG8) were each mapped to intervals <1.0 cM between two codominant markers. The effects of 21 genes in the three QTL regions were investigated by using quantitative RT-PCR analysis, and transcription profiles were obtained from the resistant TIW1 and the susceptible cSM strains. Based on transcription data, eight genes were selected for RNA interference analysis to investigate their possible roles in H. diminuta resistance, including cytochrome P450 (LOC657454) and Toll-like receptor 13 (TLR13, LOC662131). The transcription of P450 and TLR13 genes in the resistant TIW1 strains was reduced more than ninefold relative to the control. Moreover, the effects of gene knockdown of P450 and TLR13 caused resistant beetles to become susceptible to tapeworm infection, which strongly suggests an important role for each in T. castaneum resistance to H. diminuta infection.

  5. MicroRNA evolution, expression, and function during short germband development in Tribolium castaneum.

    PubMed

    Ninova, Maria; Ronshaugen, Matthew; Griffiths-Jones, Sam

    2016-01-01

    MicroRNAs are well-established players in the development of multicellular animals. Most of our understanding of microRNA function in arthropod development comes from studies in Drosophila. Despite their advantages as model systems, the long germband embryogenesis of fruit flies is an evolutionary derived state restricted to several holometabolous insect lineages. MicroRNA evolution and expression across development in animals exhibiting the ancestral and more widespread short germband mode of embryogenesis has not been characterized. We sequenced small RNA libraries of oocytes and successive intervals covering the embryonic development of the short germband model organism, Tribolium castaneum. We analyzed the evolution and temporal expression of the microRNA complement and sequenced libraries of total RNA to investigate the relationships with microRNA target expression. We show microRNA maternal loading and sequence-specific 3' end nontemplate oligoadenylation of maternally deposited microRNAs that is conserved between Tribolium and Drosophila. We further uncover large clusters encoding multiple paralogs from several Tribolium-specific microRNA families expressed during a narrow interval of time immediately after the activation of zygotic transcription. These novel microRNAs, together with several early expressed conserved microRNAs, target a significant number of maternally deposited transcripts. Comparison with Drosophila shows that microRNA-mediated maternal transcript targeting is a conserved process in insects, but the number and sequences of microRNAs involved have diverged. The expression of fast-evolving and species-specific microRNAs in the early blastoderm of T. castaneum is consistent with previous findings in Drosophila and shows that the unique permissiveness for microRNA innovation at this stage is a conserved phenomenon.

  6. MicroRNA evolution, expression, and function during short germband development in Tribolium castaneum

    PubMed Central

    Ninova, Maria; Ronshaugen, Matthew; Griffiths-Jones, Sam

    2016-01-01

    MicroRNAs are well-established players in the development of multicellular animals. Most of our understanding of microRNA function in arthropod development comes from studies in Drosophila. Despite their advantages as model systems, the long germband embryogenesis of fruit flies is an evolutionary derived state restricted to several holometabolous insect lineages. MicroRNA evolution and expression across development in animals exhibiting the ancestral and more widespread short germband mode of embryogenesis has not been characterized. We sequenced small RNA libraries of oocytes and successive intervals covering the embryonic development of the short germband model organism, Tribolium castaneum. We analyzed the evolution and temporal expression of the microRNA complement and sequenced libraries of total RNA to investigate the relationships with microRNA target expression. We show microRNA maternal loading and sequence-specific 3′ end nontemplate oligoadenylation of maternally deposited microRNAs that is conserved between Tribolium and Drosophila. We further uncover large clusters encoding multiple paralogs from several Tribolium-specific microRNA families expressed during a narrow interval of time immediately after the activation of zygotic transcription. These novel microRNAs, together with several early expressed conserved microRNAs, target a significant number of maternally deposited transcripts. Comparison with Drosophila shows that microRNA-mediated maternal transcript targeting is a conserved process in insects, but the number and sequences of microRNAs involved have diverged. The expression of fast-evolving and species-specific microRNAs in the early blastoderm of T. castaneum is consistent with previous findings in Drosophila and shows that the unique permissiveness for microRNA innovation at this stage is a conserved phenomenon. PMID:26518483

  7. Kinetic properties of alternatively spliced isoforms of laccase-2 from Tribolium castaneum and Anopheles gambiae

    PubMed Central

    Gorman, Maureen J.; Sullivan, Lucinda I.; Nguyen, Thi D. T.; Dai, Huaien; Arakane, Yasuyuki; Dittmer, Neal T.; Syed, Lateef U.; Li, Jun; Hua, Duy H.; Kanost, Michael R.

    2011-01-01

    Laccase-2 is a highly conserved multicopper oxidase that functions in insect cuticle pigmentation and tanning. In many species, alternative splicing gives rise to two laccase-2 isoforms. A comparison of laccase-2 sequences from three orders of insects revealed eleven positions at which there are conserved differences between the A and B isoforms. Homology modeling suggested that these eleven residues are not part of the substrate binding pocket. To determine whether the isoforms have different kinetic properties, we compared the activity of laccase-2 isoforms from Tribolium castaneum and Anopheles gambiae. We partially purified the four laccases as recombinant enzymes and analyzed their ability to oxidize a range of laccase substrates. The predicted endogenous substrates tested were dopamine, N-acetyldopamine (NADA), N-β-alanyldopamine (NBAD) and dopa, which were detected in T. castaneum previously and in A. gambiae as part of this study. Two additional diphenols (catechol and hydroquinone) and one non-phenolic substrate (2,2′-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid)) were also tested. We observed no major differences in substrate specificity between the A and B isoforms. Dopamine, NADA and NBAD were oxidized with catalytic efficiencies ranging from 51 – 550 min−1 mM−1. These results support the hypothesis that dopamine, NADA and NBAD are endogenous substrates for both isoforms of laccase-2. Catalytic efficiencies associated with dopa oxidation were low, ranging from 8 – 30 min−1 mM−1; in comparison, insect tyrosinase oxidized dopa with a catalytic efficiency of 201 min−1 mM−1. We found that dopa had the highest redox potential of the four endogenous substrates, and this property of dopa may explain its poor oxidation by laccase-2. We conclude that laccase-2 splice isoforms are likely to oxidize the same substrates in vivo, and additional experiments will be required to discover any isoform-specific functions. PMID:22198355

  8. Satellite DNA Modulates Gene Expression in the Beetle Tribolium castaneum after Heat Stress

    PubMed Central

    Feliciello, Isidoro; Akrap, Ivana; Ugarković, Đurđica

    2015-01-01

    Non-coding repetitive DNAs have been proposed to perform a gene regulatory role, however for tandemly repeated satellite DNA no such role was defined until now. Here we provide the first evidence for a role of satellite DNA in the modulation of gene expression under specific environmental conditions. The major satellite DNA TCAST1 in the beetle Tribolium castaneum is preferentially located within pericentromeric heterochromatin but is also dispersed as single repeats or short arrays in the vicinity of protein-coding genes within euchromatin. Our results show enhanced suppression of activity of TCAST1-associated genes and slower recovery of their activity after long-term heat stress relative to the same genes without associated TCAST1 satellite DNA elements. The level of gene suppression is not influenced by the distance of TCAST1 elements from the associated genes up to 40 kb from the genes’ transcription start sites, but it does depend on the copy number of TCAST1 repeats within an element, being stronger for the higher number of copies. The enhanced gene suppression correlates with the enrichment of the repressive histone marks H3K9me2/3 at dispersed TCAST1 elements and their flanking regions as well as with increased expression of TCAST1 satellite DNA. The results reveal transient, RNAi based heterochromatin formation at dispersed TCAST1 repeats and their proximal regions as a mechanism responsible for enhanced silencing of TCAST1-associated genes. Differences in the pattern of distribution of TCAST1 elements contribute to gene expression diversity among T. castaneum strains after long-term heat stress and might have an impact on adaptation to different environmental conditions. PMID:26275223

  9. PDP1 regulates energy metabolism through the IIS-TOR pathway in the red flour beetle, Tribolium castaneum.

    PubMed

    Li, Zhiqian; Jiang, Jianhao; Chen, Yazhou; You, Lang; Huang, Yongping; Tan, Anjiang; Li, Zhiqian; Jiang, Jianhao; Niu, Baolong; Meng, Zhiqi

    2014-03-01

    The PAR-domain protein 1 (PDP1) is essential for locomotor activity of insects. However, its functions in insect growth and development have not been studied extensively, which prompted our hypothesis that PDP1 acts in energy metabolism. Here we report identification of TcPDP1 in the red flour beetle, Tribolium castaneum, and its functional analysis by RNAi. Treating larvae with dsTcPDP1 induced pupae developmental arrestment, accompanied by accelerated fat body degradation. dsTcPDP1 treatments in adults resulted in reduced female fecundity. Disruption of TcPDP1 expression affected the transcription of genes involved in insulin signaling transduction and mechanistic target of rapamycin (mTOR) pathway. These results support our hypothesis that TcPDP1 acts in energy metabolism in T. castaneum.

  10. An unusual barrier to gene flow: perpetually immature larvae from inter-population crosses in the flour beetle, Tribolium castaneum.

    PubMed

    Drury, D W; Jideonwo, V N; Ehmke, R C; Wade, M J

    2011-12-01

    We genetically characterize an unusual hybrid incompatibility phenotype manifest in F(1) offspring of crosses between two populations of Tribolium castaneum. Hybrid larvae cease development at the third larval instar, persisting as 'perpetually immature larvae' thereafter. Although unable to produce viable adult hybrid offspring with one another, each population produces abundant, fertile hybrids with other populations, indicating a recent origin of the incompatibility and facilitating genetic studies. We mapped the paternal component of the hybrid phenotype to a single region, which exhibits two characteristics common to hybrid incompatibility: marker transmission ratio distortion within crosses and elevated genetic divergence between populations. The incompatible variation and an elevation in between-population genetic divergence is associated with a region containing the T. castaneum ecdysone receptor homologue, a major regulatory switch, controlling larval moults, pupation and metamorphosis. This contributes to understanding the genetics of speciation in the Coleoptera, one of the most speciose of all arthropod taxa.

  11. Evolutionary variations in the expression of dorso-ventral patterning genes and the conservation of pioneer neurons in Tribolium castaneum.

    PubMed

    Biffar, Lucia; Stollewerk, Angelika

    2015-04-01

    Insects are ideally suited for gaining insight into the evolutionary developmental mechanisms that have led to adaptive changes of the nervous system since the specific structure of the nervous system can be directly linked to the neural stem cell (neuroblast) lineages, which in turn can be traced back to the last common ancestor of insects. The recent comparative analysis of the Drosophila melanogaster and Tribolium castaneum neuroblast maps revealed substantial differences in the expression profiles of neuroblasts. Here we show that despite the overall conservation of the dorso-ventral expression domains of muscle segment homeobox, intermediate neuroblasts defective and ventral nervous system defective, the expression of these genes relative to the neuroblasts in the respective domains has changed considerably during insect evolution. Furthermore, functional studies show evolutionary changes in the requirement of ventral nervous system defective in the formation of neuroblast 1-1 and the correct differentiation of its presumptive progeny, the pioneer neurons aCC and pCC. The inclusion of the expression data of the dorso-ventral genes into the recently established T. castaneum neuroblast map further increases the differences in the neuroblast expression profiles between D. melanogaster and T. castaneum. Despite these molecular variations, the Even-skipped positive pioneer neurons show an invariant arrangement, except for an additional Even-skipped positive cluster that we discovered in T. castaneum. Given the importance of these pioneer neurons in establishing the intersegmental nerves and the longitudinal tracts, which are part of the conserved axonal scaffold of arthropods, we discuss internal buffering mechanisms that might ensure that neuroblast lineages invariantly generate pioneer neurons over a wide range of molecular variations.

  12. Diagnostic molecular markers for phosphine resistance in U.S. populations of Tribolium castaneum and Rhyzopertha dominica.

    PubMed

    Chen, Zhaorigetu; Schlipalius, David; Opit, George; Subramanyam, Bhadriraju; Phillips, Thomas W

    2015-01-01

    Stored product beetles that are resistant to the fumigant pesticide phosphine (hydrogen phosphide) gas have been reported for more than 40 years in many places worldwide. Traditionally, determination of phosphine resistance in stored product beetles is based on a discriminating dose bioassay that can take up to two weeks to evaluate. We developed a diagnostic cleaved amplified polymorphic sequence method, CAPS, to detect individuals with alleles for strong resistance to phosphine in populations of the red flour beetle, Tribolium castaneum, and the lesser grain borer, Rhyzopertha dominica, according to a single nucleotide mutation in the dihydrolipoamide dehydrogenase (DLD) gene. We initially isolated and sequenced the DLD genes from susceptible and strongly resistant populations of both species. The corresponding amino acid sequences were then deduced. A single amino acid mutation in DLD in populations of T. castaneum and R. dominica with strong resistance was identified as P45S in T. castaneum and P49S in R. dominica, both collected from northern Oklahoma, USA. PCR products containing these mutations were digested by the restriction enzymes MboI and BstNI, which revealed presence or absence, respectively of the resistant (R) allele and allowed inference of genotypes with that allele. Seven populations of T. castaneum from Kansas were subjected to discriminating dose bioassays for the weak and strong resistance phenotypes. Application of CAPS to these seven populations confirmed the R allele was in high frequency in the strongly resistant populations, and was absent or at a lower frequency in populations with weak resistance, which suggests that these populations with a low frequency of the R allele have the potential for selection of the strong resistance phenotype. CAPS markers for strong phosphine resistance will help to detect and confirm resistant beetles and can facilitate resistance management actions against a given pest population.

  13. Effect of ozone on respiration of adult Sitophilus oryzae (L.), Tribolium castaneum (Herbst) and Rhyzopertha dominica (F.).

    PubMed

    Lu, Baoqian; Ren, Yonglin; Du, Yu-zhou; Fu, Yueguan; Gu, Jie

    2009-10-01

    The effect of ozone on the respiration of three species of adult stored-product Coleoptera was tested in an air-tight flask. Sitophilus oryzae (L.), Rhyzopertha dominica (F.) and Tribolium castaneum (Herbst) adults were exposed to atmosphere containing 0.1, 0.2 or 0.4microg/ml initial ozone at 23-25 degrees C and 50% r.h. Carbon dioxide (CO(2)) production reflected the respiration rates of insects and was determined with a gas chromatograph (GC). The experiments showed that the effects of ozone on respiration had two distinct phases. Phase 1 involved a lower respiration rate of the adult stored-product Coleoptera under ozone atmosphere and reflected the need for insects to reduce ozone toxicity. After 1h, CO(2) production of S. oryzae was 3.19, 2.63, 2.27 and 1.99microl/mg for the ozone concentration of 0, 0.1, 0.2 and 0.4microg/ml, respectively. The results also showed that there were decreases in the rate of respiration in R. dominica and T. castaneum with an increase in ozone concentration. During phase 2, respiration of S. oryzae, R. dominica, and T. castaneum adults treated with ozone increased as the ozone degraded to oxygen. After 7h, the effect of ozone on CO(2) production, relative to the control, changed from a decrease to an increase. The findings in relation to control strategies were discussed.

  14. Genetic Variation Segregating in Natural Populations of Tribolium Castaneum Affecting Traits Observed in Hybrids with T. Freemani

    PubMed Central

    Wade, M. J.; Johnson, N. A.; Jones, R.; Siguel, V.; McNaughton, M.

    1997-01-01

    We investigated patterns of within-species genetic variation for traits observed in hybrids (hybrid numbers, hybrid sex ratios, and hybrid male deformities) between two species of flour beetles, Tribolium castaneum and T. freemani. We found genetic variation segregating among four natural populations of T. castaneum as well as within these populations. For some hybrid traits, we observed as much variation among populations 750 km apart as between populations on different continents, suggesting genetic differentiation at a local scale. Within natural populations, the variation segregating among sires is greater than that found in an earlier study for an outbred laboratory population and comparable to that observed between inbred lines derived from the outbred stock by eight generations of brother-sister mating. When sires from T. castaneum are mated to conspecific and heterospecific females, we do not observe a significant correlation at the level of the family mean between the intraspecific and interspecific phenotypes, suggesting the independence of the hybrid traits from comparable traits within species. We discuss our findings in relation to the evolutionary genetics of speciation and the expression of epistatic genetic variance in interspecific crosses. PMID:9383066

  15. Glycogen and glucose metabolism are essential for early embryonic development of the red flour beetle Tribolium castaneum.

    PubMed

    Fraga, Amanda; Ribeiro, Lupis; Lobato, Mariana; Santos, Vitória; Silva, José Roberto; Gomes, Helga; da Cunha Moraes, Jorge Luiz; de Souza Menezes, Jackson; de Oliveira, Carlos Jorge Logullo; Campos, Eldo; da Fonseca, Rodrigo Nunes

    2013-01-01

    Control of energy metabolism is an essential process for life. In insects, egg formation (oogenesis) and embryogenesis is dependent on stored molecules deposited by the mother or transcribed later by the zygote. In oviparous insects the egg becomes an isolated system after egg laying with all energy conversion taking place during embryogenesis. Previous studies in a few vector species showed a strong correlation of key morphogenetic events and changes in glucose metabolism. Here, we investigate glycogen and glucose metabolism in the red flour beetle Tribolium castaneum, an insect amenable to functional genomic studies. To examine the role of the key enzymes on glycogen and glucose regulation we cloned and analyzed the function of glycogen synthase kinase 3 (GSK-3) and hexokinase (HexA) genes during T. castaneum embryogenesis. Expression analysis via in situ hybridization shows that both genes are expressed only in the embryonic tissue, suggesting that embryonic and extra-embryonic cells display different metabolic activities. dsRNA adult female injection (parental RNAi) of both genes lead a reduction in egg laying and to embryonic lethality. Morphological analysis via DAPI stainings indicates that early development is impaired in Tc-GSK-3 and Tc-HexA1 RNAi embryos. Importantly, glycogen levels are upregulated after Tc-GSK-3 RNAi and glucose levels are upregulated after Tc-HexA1 RNAi, indicating that both genes control metabolism during embryogenesis and oogenesis, respectively. Altogether our results show that T. castaneum embryogenesis depends on the proper control of glucose and glycogen.

  16. Penetration through the peritrophic matrix is a key to lectin toxicity against Tribolium castaneum.

    PubMed

    Walski, Tomasz; Van Damme, Els J M; Smagghe, Guy

    2014-11-01

    In the last decades lectins have received a lot of attention as potential tools in pest control. Despite substantial progress in the field not all the factors determining insecticidal potency and selectivity of these proteins have been described. Recently, three lectins, RSA (Rhizoctonia solani agglutinin), SNA-I and SNA-II (Sambucus nigra agglutinin I and II) have been shown to be toxic to aphids and caterpillars. In this project we investigated if these lectins are also toxic against larvae and a cell line of the red flour beetle, Tribolium castaneum, a model organism and important pest of stored products. Furthermore, we analyzed the stability of the lectins in the larval gut and used confocal microscopy to compare their efficiency in passing through the peritrophic matrix (PM). We observed that all three lectins were toxic against the T. castaneum cell line and their effectiveness in vitro was in decreasing order SNA-II>SNA-I>RSA with the respective EC50 being 0.1, 0.5 and 3.6 μg/ml. Larvae feeding for 16 day on diets containing 2% RSA, 2% SNA-II and 2% SNA-I weighed 0.14 ± 0.07 mg, 0.67 ± 0.44 mg and 1.89 ± 0.38 mg, corresponding to approximately 7%, 36% and 80% of control larvae, respectively. As a consequence, RSA increased the time to adult emergence by over 3-fold, SNA-II by 1.9-fold and SNA-I by 1.2-fold. RSA and SNA-II were stable in the larval gut, while SNA-I was digested and excreted with the feces. Finally, confocal microscopy confirmed that RSA passed through the PM more efficiently than SNA-II. In conclusion, our data suggest that the lectin ability to pass through the PM, governed by molecule dimensions, charge and size of PM pores, is one of the features that determine the toxicity of these insecticidal proteins.

  17. Efficacies of spinosad and a combination of chlorpyrifos-methyl and deltamethrin against phosphine-resistant Rhyzopertha dominica (Coleoptera: Bostrichidae) and Tribolium castaneum (Coleoptera: Tenebrionidae) on wheat.

    PubMed

    Bajracharya, N S; Opit, George P; Talley, J; Jones, C L

    2013-10-01

    Highly phosphine-resistant populations of Rhyzopertha dominica (F.) (Coleoptera: Bostrichidae) and Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae) have recently been found in Oklahoma grain storage facilities. These findings necessitate development of a phosphine resistance management strategy to ensure continued effective use of phosphine. Therefore, we investigated the efficacies of two grain insecticides, namely, spinosad applied at label rate of 1 ppm and a mixture of chlorpyrifos-methyl and deltamethrin applied at label rates of 3 and 0.5 ppm, respectively, against highly phosphine-resistant R. dominica and T. castaneum. Adult mortality and progeny production suppression of spinosad- or chlorpyrifos-methyl + deltamethrin mixture-treated wheat that had been stored for 2, 84, 168, 252, and 336 d posttreatment were assessed. We found that both spinosad and chlorpyrifos-methyl + deltamethrin were effective against phosphine-resistant R. dominica and caused 83-100% mortality and also caused total progeny production suppression for all storage periods. Spinosad was not effective against phosphine-resistant T. castaneum; the highest mortality observed was only 3% for all the storage periods. Chlorpyrifos-methyl + deltamethrin was effective against phosphine-resistant T. castaneum only in treated wheat stored for 2 and 84 d, where it caused 93-99% mortality. However, chlorpyrifos-methyl + deltamethrin was effective and achieved total suppression of progeny production in T. castaneum for all the storage periods. Spinosad was not as effective as chlorpyrifos-methyl + deltamethrin mixture at suppressing progeny production of phosphine-resistant T. castaneum. These two insecticides can be used in a phosphine resistance management strategy for R. dominica and T. castaneum in the United States.

  18. Inbreeding depresses sperm competitiveness, but not fertilization or mating success in male Tribolium castaneum.

    PubMed

    Michalczyk, Lukasz; Martin, Oliver Y; Millard, Anna L; Emerson, Brent C; Gage, Matthew J G

    2010-11-22

    As populations decline to levels where reproduction among close genetic relatives becomes more probable, subsequent increases in homozygous recessive deleterious expression and/or loss of heterozygote advantage can lead to inbreeding depression. Here, we measure how inbreeding across replicate lines of the flour beetle Tribolium castaneum impacts on male reproductive fitness in the absence or presence of male-male competition. Effects on male evolution from mating pattern were removed by enforcing monogamous mating throughout. After inbreeding across eight generations, we found that male fertility in the absence of competition was unaffected. However, we found significant inbreeding depression of sperm competitiveness: non-inbred males won 57 per cent of fertilizations in competition, while inbred equivalents only sired 42 per cent. We also found that the P(2) 'offence' role in sperm competition was significantly more depressed under inbreeding than sperm 'defence' (P(1)). Mating behaviour did not explain these differences, and there was no difference in the viability of offspring sired by inbred or non-inbred males. Sperm length variation was significantly greater in the ejaculates of inbred males. Our results show that male ability to achieve normal fertilization success was not depressed under strong inbreeding, but that inbreeding depression in these traits occurred when conditions of sperm competition were generated. PMID:20554548

  19. Inbreeding depresses sperm competitiveness, but not fertilization or mating success in male Tribolium castaneum

    PubMed Central

    Michalczyk, Łukasz; Martin, Oliver Y.; Millard, Anna L.; Emerson, Brent C.; Gage, Matthew J. G.

    2010-01-01

    As populations decline to levels where reproduction among close genetic relatives becomes more probable, subsequent increases in homozygous recessive deleterious expression and/or loss of heterozygote advantage can lead to inbreeding depression. Here, we measure how inbreeding across replicate lines of the flour beetle Tribolium castaneum impacts on male reproductive fitness in the absence or presence of male–male competition. Effects on male evolution from mating pattern were removed by enforcing monogamous mating throughout. After inbreeding across eight generations, we found that male fertility in the absence of competition was unaffected. However, we found significant inbreeding depression of sperm competitiveness: non-inbred males won 57 per cent of fertilizations in competition, while inbred equivalents only sired 42 per cent. We also found that the P2 ‘offence’ role in sperm competition was significantly more depressed under inbreeding than sperm ‘defence’ (P1). Mating behaviour did not explain these differences, and there was no difference in the viability of offspring sired by inbred or non-inbred males. Sperm length variation was significantly greater in the ejaculates of inbred males. Our results show that male ability to achieve normal fertilization success was not depressed under strong inbreeding, but that inbreeding depression in these traits occurred when conditions of sperm competition were generated. PMID:20554548

  20. The beetle Tribolium castaneum has a fushi tarazu homolog expressed in stripes during segmentation

    NASA Technical Reports Server (NTRS)

    Brown, S. J.; Hilgenfeld, R. B.; Denell, R. E.; Spooner, B. S. (Principal Investigator)

    1994-01-01

    The genetic control of embryonic organization is far better understood for the fruit fly Drosophila melanogaster than for any other metazoan. A gene hierarchy acts during oogenesis and embryogenesis to regulate the establishment of segmentation along the anterior-posterior axis, and homeotic selector genes define developmental commitments within each parasegmental unit delineated. One of the most intensively studied Drosophila segmentation genes is fushi tarazu (ftz), a pair-rule gene expressed in stripes that is important for the establishment of the parasegmental boundaries. Although ftz is flanked by homeotic selector genes conserved throughout the metazoa, there is no evidence that it was part of the ancestral homeotic complex, and it has been unclear when the gene arose and acquired a role in segmentation. We show here that the beetle Tribolium castaneum has a ftz homolog located in its Homeotic complex and expressed in a pair-rule fashion, albeit in a register differing from that of the fly gene. These and other observations demonstrate that a ftz gene preexisted the radiation of holometabolous insects and suggest that it has a role in beetle embryogenesis which differs somewhat from that described in flies.

  1. Direct and indirect genetic effects in life-history traits of flour beetles (Tribolium castaneum).

    PubMed

    Ellen, Esther D; Peeters, Katrijn; Verhoeven, Merel; Gols, Rieta; Harvey, Jeffrey A; Wade, Michael J; Dicke, Marcel; Bijma, Piter

    2016-01-01

    Indirect genetic effects (IGEs) are the basis of social interactions among conspecifics, and can affect genetic variation of nonsocial and social traits. We used flour beetles (Tribolium castaneum) of two phenotypically distinguishable populations to estimate genetic (co)variances and the effect of IGEs on three life-history traits: development time (DT), growth rate (GR), and pupal body mass (BM). We found that GR was strongly affected by social environment with IGEs accounting for 18% of the heritable variation. We also discovered a sex-specific social effect: male ratio in a group significantly affected both GR and BM; that is, beetles grew larger and faster in male-biased social environments. Such sex-specific IGEs have not previously been demonstrated in a nonsocial insect. Our results show that beetles that achieve a higher BM do so via a slower GR in response to social environment. Existing models of evolution in age-structured or stage-structured populations do not account for IGEs of social cohorts. It is likely that such IGEs have played a key role in the evolution of developmental plasticity shown by Tenebrionid larvae in response to density. Our results document an important source of genetic variation for GR, often overlooked in life-history theory. PMID:26660947

  2. Insect gravitational biology: ground-based and shuttle flight experiments using the beetle Tribolium castaneum

    NASA Technical Reports Server (NTRS)

    Bennett, R. L.; Abbott, M. K.; Denell, R. E.; Spooner, B. S. (Principal Investigator)

    1994-01-01

    Many of the traditional experimental advantages of insects recommend their use in studies of gravitational and space biology. The fruit fly, Drosophila melanogaster, is an obvious choice for studies of the developmental significance of gravity vectors because of the unparalleled description of regulatory mechanisms controlling oogenesis and embryogenesis. However, we demonstrate that Drosophila could not survive the conditions mandated for particular flight opportunities on the Space Shuttle. With the exception of Drosophila, the red flour beetle, Tribolium castaneum, is the insect best characterized with respect to molecular embryology and most frequently utilized for past space flights. We show that Tribolium is dramatically more resistant to confinement in small sealed volumes. In preparation for flight experiments we characterize the course and timing of the onset of oogenesis in newly eclosed adult females. Finally, we present results from two shuttle flights which indicate that a number of aspects of the development and function of the female reproductive system are not demonstrably sensitive to microgravity. Available information supports the utility of this insect for future studies of gravitational biology.

  3. Satellite DNA as a driver of population divergence in the red flour beetle Tribolium castaneum.

    PubMed

    Feliciello, Isidoro; Akrap, Ivana; Brajković, Josip; Zlatar, Ivo; Ugarković, Đurđica

    2014-12-19

    Tandemly repeated satellite DNAs are among most rapidly evolving sequences in eukaryotic genome, usually differing significantly among closely related species. By inducing changes in heterochromatin and/or centromere, satellite DNAs are expected to drive population and species divergence. However, despite high evolutionary dynamics, divergence of satellite DNA profiles at the level of natural population which precedes and possibly triggers speciation process is not readily detected. Here, we characterize minor TCAST2 satellite DNA of the red flour beetle Tribolium castaneum and follow its dynamics among wild-type strains originating from diverse geographic locations. The investigation revealed presence of three distinct subfamilies of TCAST2 satellite DNA which differ in monomer size, genome organization, and subfamily specific mutations. Subfamilies Tcast2a and Tcast2b are tandemly arranged within pericentromeric heterochromatin whereas Tcast2c is preferentially dispersed within euchromatin of all chromosomes. Among strains, TCAST2 subfamilies are conserved in sequence but exhibit a significant content variability. This results in overrepresentation or almost complete absence of particular subfamily in some strains and enables discrimination between strains. It is proposed that homologous recombination, probably stimulated by environmental stress, is responsible for the emergence of TCAST2 satellite subfamilies, their copy number variation and dispersion within genome. The results represent the first evidence for the existence of population-specific satellite DNA profiles. Partial organization of TCAST2 satellite DNA in the form of single repeats dispersed within euchromatin additionally contributes to the genome divergence at the population level.

  4. Novel functions for Dorsocross in epithelial morphogenesis in the beetle Tribolium castaneum.

    PubMed

    Horn, Thorsten; Panfilio, Kristen A

    2016-08-15

    Epithelial morphogenesis, the progressive restructuring of tissue sheets, is fundamental to embryogenesis. In insects, not only embryonic tissues but also extraembryonic (EE) epithelia play a crucial role in shaping the embryo. In Drosophila, the T-box transcription factor Dorsocross (Doc) is essential for EE tissue maintenance and therefore embryo survival. However, Drosophila possesses a single amnioserosa, whereas most insects have a distinct amnion and serosa. How does this derived situation compare with Doc function in the ancestral context of two EE epithelia? Here, we investigate the Doc orthologue in the red flour beetle, Tribolium castaneum, which is an excellent model for EE tissue complement and for functional, fluorescent live imaging approaches. Surprisingly, we find that Tc-Doc controls all major events in Tribolium EE morphogenesis without affecting EE tissue specification or maintenance. These macroevolutionary changes in function between Tribolium and Drosophila are accompanied by regulatory network changes, where BMP signaling and possibly the transcription factor Hindsight are downstream mediators. We propose that the ancestral role of Doc was to control morphogenesis and discuss how Tc-Doc could provide spatial precision for remodeling the amnion-serosa border. PMID:27407103

  5. Insulin receptor regulates food intake through sulfakinin signaling in the red flour beetle, Tribolium castaneum.

    PubMed

    Lin, Xianyu; Yu, Na; Smagghe, Guy

    2016-06-01

    Insects obtain energy and nutrients via feeding to support growth and development. The insulin signaling pathway is involved in the regulation of feeding; however, the underlying mechanisms are not fully understood. Here, we show that insulin signaling regulates food intake via crosstalk with neuropeptide sulfakinin in the red flour beetle, Tribolium castaneum. Silencing of the insulin receptor (InR) decreased the food intake in the penultimate and final instar stages, leading to a decrease of weight gain and mortality during larval-pupal metamorphosis. Interestingly, the knockdown of InR co-occurred with an increased expression of sulfakinin (sk), a gene encoding neuropeptide SK functioning as a satiety signal. In parallel, double silencing of sk and InR eliminated the inhibitory effect on food intake as induced by silencing of InR and the larvae died as prepupae. In conclusion, this study shows, for the first time, that the insulin/InR signaling regulates food intake through the sulfakinin signaling pathway in the larval stages of this important model and pest insect, indicating a novel target for pest control. PMID:26972481

  6. Satellite DNA as a Driver of Population Divergence in the Red Flour Beetle Tribolium castaneum

    PubMed Central

    Feliciello, Isidoro; Akrap, Ivana; Brajković, Josip; Zlatar, Ivo; Ugarković, Đurđica

    2015-01-01

    Tandemly repeated satellite DNAs are among most rapidly evolving sequences in eukaryotic genome, usually differing significantly among closely related species. By inducing changes in heterochromatin and/or centromere, satellite DNAs are expected to drive population and species divergence. However, despite high evolutionary dynamics, divergence of satellite DNA profiles at the level of natural population which precedes and possibly triggers speciation process is not readily detected. Here, we characterize minor TCAST2 satellite DNA of the red flour beetle Tribolium castaneum and follow its dynamics among wild-type strains originating from diverse geographic locations. The investigation revealed presence of three distinct subfamilies of TCAST2 satellite DNA which differ in monomer size, genome organization, and subfamily specific mutations. Subfamilies Tcast2a and Tcast2b are tandemly arranged within pericentromeric heterochromatin whereas Tcast2c is preferentially dispersed within euchromatin of all chromosomes. Among strains, TCAST2 subfamilies are conserved in sequence but exhibit a significant content variability. This results in overrepresentation or almost complete absence of particular subfamily in some strains and enables discrimination between strains. It is proposed that homologous recombination, probably stimulated by environmental stress, is responsible for the emergence of TCAST2 satellite subfamilies, their copy number variation and dispersion within genome. The results represent the first evidence for the existence of population-specific satellite DNA profiles. Partial organization of TCAST2 satellite DNA in the form of single repeats dispersed within euchromatin additionally contributes to the genome divergence at the population level. PMID:25527837

  7. Constraints on the evolution of function-valued traits: a study of growth in Tribolium castaneum.

    PubMed

    Irwin, K K; Carter, P A

    2013-12-01

    Growth trajectories often impact individual fitness. They are continuous by nature and so are amenable to analysis using a function-valued (FV) trait framework to reveal their underlying genetic architecture. Previous studies have found high levels of standing additive genetic (co)variance for growth trajectories despite the expectation that growth should be responding to frequent strong directional selection. In this study, the FV framework is used to estimate the additive genetic covariance function for growth trajectories in larval Tribolium castaneum to address questions about standing additive genetic (co)variance and possible evolutionary constraints on growth and to predict responses to four plausible selection regimes. Results show that additive genetic (co)variance is high at the early ages, but decreases towards later ages in the larval period. A selection gradient function of the same size and in the same direction of the first eigenfunction of the G-function should give the maximal response. However, evolutionary constraints may be acting to keep this maximal response from being realized, through either conflicting effects on survivability and fecundity of larger body size, few evolutionary directions having sufficient additive variance for a response, genetic trade-offs with other traits or physiological regulatory mechanisms. More light may be shed on these constraints through the development of more sophisticated statistical approaches and implementation of additional empirical studies to explicitly test for specific types of constraints.

  8. Evaluation of synergized pyrethrin aerosol for control of Tribolium castaneum and Tribolium confusum (Coleoptera: Tenebrionidae).

    PubMed

    Kharel, Kabita; Arthur, Frank H; Zhu, Kun Yan; Campbell, James F; Subramanyam, Bhadriraju

    2014-02-01

    Aerosol insecticides are being used in flour mill pest management programs, but there is limited information on their efficacy on different insect life stages. In this study, we evaluated the efficacy of synergized pyrethrin applied as an aerosol against eggs, larvae, pupae, and adults of the red flour beetle, Tribolium castaneum (Herbst), and the confused flour beetle, Tribolium confusum Jacquelin du Val. Effects of direct and indirect exposure were evaluated by exposing each life stage to the aerosol and then transferring to untreated flour, transferring untreated insects to treated flour, or exposing both the insects and the flour to the aerosol. The aerosol produced >88% mortality of both species and all life stages when insects were directly treated and transferred to either treated or untreated flour. Mortality was significantly reduced when insects were either treated together with flour or untreated insects were transferred to treated flour (indirect exposure to the aerosol). Larvae and adults of both species were more tolerant compared with eggs and pupae. Recovery of moribund adults in the indirect exposure treatments was greater compared with recovery of moribund insects in the direct exposure treatments. Good sanitation before aerosol application could facilitate direct exposure of insects and thus increase aerosol efficacy inside flour mills.

  9. Hormonal enhancement of insecticide efficacy in Tribolium castaneum: oxidative stress and metabolic aspects.

    PubMed

    Plavšin, Ivana; Stašková, Tereza; Šerý, Michal; Smýkal, Vlastimil; Hackenberger, Branimir K; Kodrík, Dalibor

    2015-04-01

    Insect anti-stress responses, including those induced by insecticides, are controlled by adipokinetic hormones (AKHs). We examined the physiological consequences of Pyrap-AKH application on Tribolium castaneum adults (AKH-normal and AKH-deficient prepared by the RNAi technique) treated by two insecticides, pirimiphos-methyl and deltamethrin. Co-application of pirimiphos-methyl and/or deltamethrin with AKH significantly increased beetle mortality compared with application of the insecticides alone. This co-treatment was accompanied by substantial stimulation of general metabolism, as monitored by carbon dioxide production. Further, the insecticide treatment alone affected some basic markers of oxidative stress: it lowered total antioxidative capacity as well as the activity of superoxide dismutase in the beetle body; in addition, it enhanced the activity of catalase and glutathione-S-transferase. However, these discrepancies in oxidative stress markers were eliminated/reduced by co-application with Pyrap-AKH. We suggest that the elevation of metabolism, which is probably accompanied with faster turnover of toxins, might be responsible for the higher mortality that results after AKH and insecticide co-application. Changes in oxidative stress markers are probably not included in the mechanisms responsible for increased mortality.

  10. The Tribolium castaneum cell line TcA: a new tool kit for cell biology.

    PubMed

    Silver, Kristopher; Jiang, Hongbo; Fu, Jinping; Phillips, Thomas W; Beeman, Richard W; Park, Yoonseong

    2014-10-30

    The red flour beetle, Tribolium castaneum, is an agriculturally important insect pest that has been widely used as a model organism. Recently, an adherent cell line (BCIRL-TcA-CLG1 or TcA) was developed from late pupae of the red flour beetle. Next generation transcriptome sequencing of TcA cells demonstrated expression of a wide variety of genes associated with specialized functions in chitin metabolism, immune responses and cellular and systemic RNAi pathways. Accordingly, we evaluated the sensitivity of TcA cells to dsRNA to initiate an RNAi response. TcA cells were highly sensitive to minute amounts of dsRNA, with a minimum effective dose of 100 pg/mL resulting in significant suppression of gene expression. We have also developed a plasmid containing two TcA-specific promoters, the promoter from the 40S ribosomal protein subunit (TC006550) and a bi-directional heat shock promoter (TcHS70) from the intergenic space between heat shock proteins 68a and b. These promoters have been employed to provide high levels of either constitutive (TC006550) or inducible (TcHS70) gene expression of the reporter proteins. Our results show that the TcA cell line, with its sensitivity to RNAi and functional TcA-specific promoters, is an invaluable resource for studying basic molecular and physiological questions.

  11. Are there genetic trade-offs between immune and reproductive investments in Tribolium castaneum?

    PubMed

    Hangartner, Sandra; Sbilordo, Sonja H; Michalczyk, Łukasz; Gage, Matthew J G; Martin, Oliver Y

    2013-10-01

    Parasites impose strong selection on hosts to defend themselves, which is expected to result in trade-offs with other fitness traits such as reproduction. Here we test for genetic trade-offs between reproductive traits and immunity using Tribolium castaneum lines that were subject to experimental evolution. The lines have been exposed to contrasting sexual selection intensities via different sex ratios (female-biased, equal and male-biased). After 56 generations, the lines have significantly diverged and those experiencing high sexual selection have evolved males who are superior competitors for reproductive success, and females who are more resistant to multiple mating. All selected lines were assessed for both an immune measurement (phenoloxidase (PO) activity) and host resistance to the microsporidian Nosema whitei after two generations of relaxed selection. In contrast to our expectations we did not find any evidence for a genetic trade-off between investment in reproduction and immunity. Both PO and Nosema resistance did not differ between lines, despite their divergences in reproductive investment due to variation in sexual selection and conflict. Nevertheless, overall we found that females had higher PO activities and in the Nosema free control treatment survived longer than males, suggesting that females generally invest more in PO and survival under control conditions than males. This result fits the Bateman's principle, which states that females gain fitness through increased immunity and longevity, while males gain fitness through increased mating success.

  12. Direct and indirect genetic effects in life-history traits of flour beetles (Tribolium castaneum).

    PubMed

    Ellen, Esther D; Peeters, Katrijn; Verhoeven, Merel; Gols, Rieta; Harvey, Jeffrey A; Wade, Michael J; Dicke, Marcel; Bijma, Piter

    2016-01-01

    Indirect genetic effects (IGEs) are the basis of social interactions among conspecifics, and can affect genetic variation of nonsocial and social traits. We used flour beetles (Tribolium castaneum) of two phenotypically distinguishable populations to estimate genetic (co)variances and the effect of IGEs on three life-history traits: development time (DT), growth rate (GR), and pupal body mass (BM). We found that GR was strongly affected by social environment with IGEs accounting for 18% of the heritable variation. We also discovered a sex-specific social effect: male ratio in a group significantly affected both GR and BM; that is, beetles grew larger and faster in male-biased social environments. Such sex-specific IGEs have not previously been demonstrated in a nonsocial insect. Our results show that beetles that achieve a higher BM do so via a slower GR in response to social environment. Existing models of evolution in age-structured or stage-structured populations do not account for IGEs of social cohorts. It is likely that such IGEs have played a key role in the evolution of developmental plasticity shown by Tenebrionid larvae in response to density. Our results document an important source of genetic variation for GR, often overlooked in life-history theory.

  13. Gender- and stressor-specific microRNA expression in Tribolium castaneum.

    PubMed

    Freitak, Dalial; Knorr, Eileen; Vogel, Heiko; Vilcinskas, Andreas

    2012-10-23

    MicroRNAs (miRNAs) are small non-coding RNAs mediating post-transcriptional regulation of gene expression in eukaryotes. Addressing their role in regulation of physiological adaptations to environmental stress in insects, we selected the red flour beetle Tribolium castaneum as a model. Beetles were fed with the bacterial entomopathogen Pseudomonas entomophila (to mimic natural infection), injected with peptidoglycan (experimental setting of strong immune responses) or subjected to either mild heat shock or starvation. Differential expression of selected immunity- and stress-related genes was quantified using real-time PCR, and expression and induction of 455 mature arthropod miRNAs were determined using proprietary microarrays. We found that Tribolium exhibits both gender- and stressor-specific adjustment of immune gene and miRNA expression. Strikingly, we discovered that the number of stressor-induced miRNAs in females is remarkably higher than in males. This observation could support the hypothesis called Bateman's principle in immunity that predicts gender-specific immune responses because females gain fitness through increased longevity, whereas males gain fitness by increasing mating rates. Our results suggest that Tribolium males and females display differential regulatory elements, both pre- and post-transcriptional, likely resulting from different investment strategies in life-history traits.

  14. Functional value of elytra under various stresses in the red flour beetle, Tribolium castaneum

    PubMed Central

    Linz, David M.; Hu, Alan W.; Sitvarin, Michael I.; Tomoyasu, Yoshinori

    2016-01-01

    Coleoptera (beetles) is a massively successful order of insects, distinguished by their evolutionarily modified forewings called elytra. These structures are often presumed to have been a major driving force for the successful radiation of this taxon, by providing beetles with protection against a variety of harsh environmental factors. However, few studies have directly demonstrated the functional significance of the elytra against diverse environmental challenges. Here, we sought to empirically test the function of the elytra using Tribolium castaneum (the red flour beetle) as a model. We tested four categories of stress on the beetles: physical damage to hindwings, predation, desiccation, and cold shock. We found that, in all categories, the presence of elytra conferred a significant advantage compared to those beetles with their elytra experimentally removed. This work provides compelling quantitative evidence supporting the importance of beetle forewings in tolerating a variety of environmental stresses, and gives insight into how the evolution of elytra have facilitated the remarkable success of beetle radiation. PMID:27708390

  15. Multiple Wnt genes are required for segmentation in the short-germ embryo of Tribolium castaneum.

    PubMed

    Bolognesi, Renata; Farzana, Laila; Fischer, Tamara D; Brown, Susan J

    2008-10-28

    wingless (wg)/Wnt family are essential to development in virtually all metazoans. In short-germ insects, including the red flour beetle (Tribolium castaneum), the segment-polarity function of wg is conserved [1]. Wnt signaling is also implicated in posterior patterning and germband elongation [2-4], but despite its expression in the posterior growth zone, Wnt1/wg alone is not responsible for these functions [1-3]. Tribolium contains additional Wnt family genes that are also expressed in the growth zone [5]. After depleting Tc-WntD/8 we found a small percentage of embryos lacking abdominal segments. Additional removal of Tc-Wnt1 significantly enhanced the penetrance of this phenotype. Seeking alternative methods to deplete Wnt signal, we performed RNAi with other components of the Wnt pathway including wntless (wls), porcupine (porc), and pangolin (pan). Tc-wls RNAi caused segmentation defects similar to Tc-Wnt1 RNAi, but not Tc-WntD/8 RNAi, indicating that Tc-WntD/8 function is Tc-wls independent. Depletion of Tc-porc and Tc-pan produced embryos resembling double Tc-Wnt1,Tc-WntD/8 RNAi embryos, suggesting that Tc-porc is essential for the function of both ligands, which signal through the canonical pathway. This is the first evidence of functional redundancy between Wnt ligands in posterior patterning in short-germ insects. This Wnt function appears to be conserved in other arthropods [6] and vertebrates [7-9].

  16. Genome-wide mapping of conserved microRNAs and their host transcripts in Tribolium castaneum.

    PubMed

    Luo, Qibin; Zhou, Qing; Yu, Xiaomin; Lin, Hongbin; Hu, Songnian; Yu, Jun

    2008-06-01

    MicroRNAs (miRNAs) are endogenous 22-nt RNAs, which play important regulatory roles by post-transcriptional gene silencing. A computational strategy has been developed for the identification of conserved miRNAs based on features of known metazoan miRNAs in red flour beetle (Tribolium castaneum), which is regarded as one of the major laboratory models of arthropods. Among 118 putative miRNAs, 47% and 53% of the predicted miRNAs from the red flour beetle are harbored by known protein-coding genes (intronic) and genes located outside (intergenic miRNA), respectively. There are 31 intronic miRNAs in the same transcriptional orientation as the host genes, which may share RNA polymerase II and spliceosomal machinery with their host genes for their biogenesis. A hypothetical feedback model has been proposed based on the analysis of the relationship between intronic miRNAs and their host genes in the development of red flour beetle. PMID:18571123

  17. Comparative characterization of two intracellular Ca²⁺-release channels from the red flour beetle, Tribolium castaneum.

    PubMed

    Liu, Yaping; Li, Chengjun; Gao, Jingkun; Wang, Wenlong; Huang, Li; Guo, Xuezhu; Li, Bin; Wang, Jianjun

    2014-10-21

    Ryanodine receptors (RyRs) and inositol 1,4,5-trisphosphate receptors (IP3Rs) are members of a family of tetrameric intracellular Ca(2+)-release channels (CRCs). While it is well known in mammals that RyRs and IP3Rs modulate multiple physiological processes, the roles of these two CRCs in the development and physiology of insects remain poorly understood. In this study, we cloned and functionally characterized RyR and IP3R cDNAs (named TcRyR and TcIP3R) from the red flour beetle, Tribolium castaneum. The composite TcRyR gene contains an ORF of 15,285 bp encoding a protein of 5,094 amino acid residues. The TcIP3R contains an 8,175 bp ORF encoding a protein of 2,724 amino acids. Expression analysis of TcRyR and TcIP3R revealed significant differences in mRNA expression levels among T. castaneum during different developmental stages. When the transcript levels of TcRyR were suppressed by RNA interference (RNAi), an abnormal folding of the adult hind wings was observed, while the RNAi-mediated knockdown of TcIP3R resulted in defective larval-pupal and pupal-adult metamorphosis. These results suggested that TcRyR is required for muscle excitation-contraction (E-C) coupling in T. castaneum, and that calcium release via IP3R might play an important role in regulating ecdysone synthesis and release during molting and metamorphosis in insects.

  18. Comparative toxicity and micronuclei formation in Tribolium castaneum, Callosobruchus maculatus and Sitophilus oryzae exposed to high doses of gamma radiation.

    PubMed

    Ahmadi, Mehrdad; Mozdarani, Hossein; Abd-Alla, Adly M M

    2015-07-01

    The effects of gamma radiation on mortality and micronucleus formation in Tribolium castaneum Herbst, Callosobruchus maculatus (F.) and Sitophilus oryzae (L.) genital cells were evaluated. Two groups of healthy and active adult insects 1-3 and 8-10 days old were irradiated with various doses (50-200 Gy) gamma ray. Seven days post-irradiation; mortality rates and micronucleus formation were assessed in genital cells of the irradiated insects. The results show that with increasing gamma doses, the mortality rate of each species increased and T. castaneum and S. oryzae showed the low and high sensitivity respectively. It was shown that the micronucleus appearance in the tested insects had correlation with amount and intensity of radiation doses. Moreover our results indicate different levels in the genotoxicity of gamma radiation among the insects' genital cells under study. The frequency of micronuclei in genital cells of 1-3 days old insects exposed to 50 and 200 Gy were 12.6 and 38.8 Mn/1000 cells in T. castaneum, 20.8 and 46.8 Mn/1000 cells in C. maculatus and 16.8 and 57.2 Mn/1000 cells in S. oryzae respectively. A high sensitivity of the genital cells to irradiation exposure was seen in S. oryzae correlated with its high mortality rate compared with the other two species. These results might be indicative of inflicting chromosomal damage expressed as micronucleus in high mortality rates observed in the pest population; an indication of genotoxic effects of radiation on the studied species.

  19. Efficient CRISPR-mediated gene targeting and transgene replacement in the beetle Tribolium castaneum.

    PubMed

    Gilles, Anna F; Schinko, Johannes B; Averof, Michalis

    2015-08-15

    Gene-editing techniques are revolutionizing the way we conduct genetics in many organisms. The CRISPR/Cas nuclease has emerged as a highly versatile, efficient and affordable tool for targeting chosen sites in the genome. Beyond its applications in established model organisms, CRISPR technology provides a platform for genetic intervention in a wide range of species, limited only by our ability to deliver it to cells and to select mutations efficiently. Here, we test the CRISPR technology in an emerging insect model and pest, the beetle Tribolium castaneum. We use simple assays to test CRISPR/Cas activity, we demonstrate efficient expression of guide RNAs and Cas9 from Tribolium U6 and hsp68 promoters and we test the efficiency of knockout and knock-in approaches in Tribolium. We find that 55-80% of injected individuals carry mutations (indels) generated by non-homologous end joining, including mosaic bi-allelic knockouts; 71-100% carry such mutations in their germ line and transmit them to the next generation. We show that CRISPR-mediated gene knockout of the Tribolium E-cadherin gene causes defects in dorsal closure, which is consistent with RNAi-induced phenotypes. Homology-directed knock-in of marker transgenes was observed in 14% of injected individuals and transmitted to the next generation by 6% of injected individuals. Previous work in Tribolium mapped a large number of transgene insertions associated with developmental phenotypes and enhancer traps. We present an efficient method for re-purposing these insertions, via CRISPR-mediated replacement of these transgenes by new constructs. PMID:26160901

  20. Multiple Wnt genes are required for segmentation in the short-germ embryo of Tribolium castaneum.

    PubMed

    Bolognesi, Renata; Farzana, Laila; Fischer, Tamara D; Brown, Susan J

    2008-10-28

    wingless (wg)/Wnt family are essential to development in virtually all metazoans. In short-germ insects, including the red flour beetle (Tribolium castaneum), the segment-polarity function of wg is conserved [1]. Wnt signaling is also implicated in posterior patterning and germband elongation [2-4], but despite its expression in the posterior growth zone, Wnt1/wg alone is not responsible for these functions [1-3]. Tribolium contains additional Wnt family genes that are also expressed in the growth zone [5]. After depleting Tc-WntD/8 we found a small percentage of embryos lacking abdominal segments. Additional removal of Tc-Wnt1 significantly enhanced the penetrance of this phenotype. Seeking alternative methods to deplete Wnt signal, we performed RNAi with other components of the Wnt pathway including wntless (wls), porcupine (porc), and pangolin (pan). Tc-wls RNAi caused segmentation defects similar to Tc-Wnt1 RNAi, but not Tc-WntD/8 RNAi, indicating that Tc-WntD/8 function is Tc-wls independent. Depletion of Tc-porc and Tc-pan produced embryos resembling double Tc-Wnt1,Tc-WntD/8 RNAi embryos, suggesting that Tc-porc is essential for the function of both ligands, which signal through the canonical pathway. This is the first evidence of functional redundancy between Wnt ligands in posterior patterning in short-germ insects. This Wnt function appears to be conserved in other arthropods [6] and vertebrates [7-9]. PMID:18926702

  1. Juvenile hormone regulation of male accessory gland activity in the red flour beetle, Tribolium castaneum

    PubMed Central

    Parthasarathy, R.; Tan, A.; Sun, Z.; Chen, J.; Rainkin, M.; Palli, S. R.

    2009-01-01

    Male accessory gland proteins (Acps) act as key modulators of reproductive success in insects by influencing the female reproductive physiology and behavior. We used custom microarrays and identified 112 genes that were highly expressed in male accessory glands (MAG) in the red flour beetle, Tribolium castaneum. Out of these 112 identified genes, 59 of them contained sequences coding for signal peptide and cleavage site and the remaining 53 contained transmembrane domains. The expression of 14 these genes in the MAG but not in other tissues of male or female was confirmed by quantitative real-time PCR. In virgin males, juvenile hormone (JH) levels increased from second day post adult emergence (PAE), remained high on third day PAE and declined on fourth day PAE. The ecdysteroid titers were high soon after adult emergence but declined to minimal levels from 1-5 days PAE. Feeding of juvenile hormone analog, hydroprene, but not the ecdysteroid analog, RH-2485, showed an increase in size of MAGs, as well as an increase in total RNA and protein content of MAG. Hydroprene treatment also increased the expression Acp genes in the MAG. RNAi-mediated knock-down in the expression of JHAMT gene decreased the size of MAGs and expression of Acps. JH deficiency influenced male reproductive fitness as evidenced by a less vigor in mating behavior, poor sperm transfer, low egg and the progeny production by females mated with the JH deficient males. These data suggest a critical role for JH in the regulation of male reproduction especially through MAG secretions. PMID:19324087

  2. Anatomical and morphological spine variation in Gymnocalycium kieslingii subsp. castaneum (Cactaceae)

    PubMed Central

    Gebauer, Roman; Řepka, Radomír; Šmudla, Radek; Mamoňová, Miroslava; Ďurkovič, Jaroslav

    2016-01-01

    Abstract Although spine variation within cacti species or populations is assumed to be large, the minimum sample size of different spine anatomical and morphological traits required for species description is less studied. There are studies where only 2 spines were used for taxonomical comparison amnog species. Therefore, the spine structure variation within areoles and individuals of one population of Gymnocalycium kieslingii subsp. castaneum (Ferrari) Slaba was analyzed. Fifteen plants were selected and from each plant one areole from the basal, middle and upper part of the plant body was sampled. A scanning electron microscopy was used for spine surface description and a light microscopy for measurements of spine width, thickness, cross-section area, fiber diameter and fiber cell wall thickness. The spine surface was more visible and damaged less in the upper part of the plant body than in the basal part. Large spine and fiber differences were found between upper and lower parts of the plant body, but also within single areoles. In general, the examined traits in the upper part had by 8–17% higher values than in the lower parts. The variation of spine and fiber traits within areoles was lower than the differences between individuals. The minimum sample size was largely influenced by the studied spine and fiber traits, ranging from 1 to 70 spines. The results provide pioneer information useful in spine sample collection in the field for taxonomical, biomechanical and structural studies. Nevertheless, similar studies should be carried out for other cacti species to make generalizations. The large spine and fiber variation within areoles observed in our study indicates a very complex spine morphogenesis.

  3. Efficient CRISPR-mediated gene targeting and transgene replacement in the beetle Tribolium castaneum.

    PubMed

    Gilles, Anna F; Schinko, Johannes B; Averof, Michalis

    2015-08-15

    Gene-editing techniques are revolutionizing the way we conduct genetics in many organisms. The CRISPR/Cas nuclease has emerged as a highly versatile, efficient and affordable tool for targeting chosen sites in the genome. Beyond its applications in established model organisms, CRISPR technology provides a platform for genetic intervention in a wide range of species, limited only by our ability to deliver it to cells and to select mutations efficiently. Here, we test the CRISPR technology in an emerging insect model and pest, the beetle Tribolium castaneum. We use simple assays to test CRISPR/Cas activity, we demonstrate efficient expression of guide RNAs and Cas9 from Tribolium U6 and hsp68 promoters and we test the efficiency of knockout and knock-in approaches in Tribolium. We find that 55-80% of injected individuals carry mutations (indels) generated by non-homologous end joining, including mosaic bi-allelic knockouts; 71-100% carry such mutations in their germ line and transmit them to the next generation. We show that CRISPR-mediated gene knockout of the Tribolium E-cadherin gene causes defects in dorsal closure, which is consistent with RNAi-induced phenotypes. Homology-directed knock-in of marker transgenes was observed in 14% of injected individuals and transmitted to the next generation by 6% of injected individuals. Previous work in Tribolium mapped a large number of transgene insertions associated with developmental phenotypes and enhancer traps. We present an efficient method for re-purposing these insertions, via CRISPR-mediated replacement of these transgenes by new constructs.

  4. TrOn: an anatomical ontology for the beetle Tribolium castaneum.

    PubMed

    Dönitz, Jürgen; Grossmann, Daniela; Schild, Inga; Schmitt-Engel, Christian; Bradler, Sven; Prpic, Nikola-Michael; Bucher, Gregor

    2013-01-01

    In a morphological ontology the expert's knowledge is represented in terms, which describe morphological structures and how these structures relate to each other. With the assistance of ontologies this expert knowledge is made processable by machines, through a formal and standardized representation of terms and their relations to each other. The red flour beetle Tribolium castaneum, a representative of the most species rich animal taxon on earth (the Coleoptera), is an emerging model organism for development, evolution, physiology, and pest control. In order to foster Tribolium research, we have initiated the Tribolium Ontology (TrOn), which describes the morphology of the red flour beetle. The content of this ontology comprises so far most external morphological structures as well as some internal ones. All modeled structures are consistently annotated for the developmental stages larva, pupa and adult. In TrOn all terms are grouped into three categories: Generic terms represent morphological structures, which are independent of a developmental stage. In contrast, downstream of such terms are concrete terms which stand for a dissectible structure of a beetle at a specific life stage. Finally, there are mixed terms describing structures that are only found at one developmental stage. These terms combine the characteristics of generic and concrete terms with features of both. These annotation principles take into account the changing morphology of the beetle during development and provide generic terms to be used in applications or for cross linking with other ontologies and data resources. We use the ontology for implementing an intuitive search function at the electronic iBeetle-Base, which stores morphological defects found in a genome wide RNA interference (RNAi) screen. The ontology is available for download at http://ibeetle-base.uni-goettingen.de.

  5. Signalling properties and pharmacology of a 5-HT7 -type serotonin receptor from Tribolium castaneum.

    PubMed

    Vleugels, R; Lenaerts, C; Vanden Broeck, J; Verlinden, H

    2014-04-01

    In the last decade, genome sequence data and gene structure information on invertebrate receptors has been greatly expanded by large sequencing projects and cloning studies. This information is of great value for the identification of receptors; however, functional and pharmacological data are necessary for an accurate receptor classification and for practical applications. In insects, an important group of neurotransmitter and neurohormone receptors, for which ample sequence information is available but pharmacological information is missing, are the biogenic amine G protein-coupled receptors (GPCRs). In the present study, we investigated the sequence information, pharmacology and signalling properties of a 5-HT7 -type serotonin receptor from the red flour beetle, Tribolium castaneum (Trica5-HT7 ). The receptor encoding cDNA shows considerable sequence similarity with cognate 5-HT7 receptors and phylogenetic analysis also clusters the receptor within this 5-HT receptor group. Real-time reverse transcription PCR demonstrated high expression levels in the brain, indicating the possible importance of this receptor in neural processes. Trica5-HT7 was dose-dependently activated by 5-HT, which induced elevated intracellular cyclic AMP levels but had no effect on calcium signalling. The synthetic agonists, α-methyl 5-HT, 5-methoxytryptamine, 5-carboxamidotryptamine and 8-hydroxy-2-(dipropylamino)tetralin hydrobromide, showed a response, although with a much lower potency and efficacy than 5-HT. Ketanserin and methiothepin were the most potent antagonists. Both showed characteristics of competitive inhibition on Trica5-HT7 . The signalling pathway and pharmacological profile offer important information that will facilitate functional and comparative studies of 5-HT receptors in insects and other invertebrates. The pharmacology of invertebrate 5-HT receptors differs considerably from that of vertebrates. The present study may therefore contribute to establishing a more

  6. Non-invasive long-term fluorescence live imaging of Tribolium castaneum embryos.

    PubMed

    Strobl, Frederic; Stelzer, Ernst H K

    2014-06-01

    Insect development has contributed significantly to our understanding of metazoan development. However, most information has been obtained by analyzing a single species, the fruit fly Drosophila melanogaster. Embryonic development of the red flour beetle Tribolium castaneum differs fundamentally from that of Drosophila in aspects such as short-germ development, embryonic leg development, extensive extra-embryonic membrane formation and non-involuted head development. Although Tribolium has become the second most important insect model organism, previous live imaging attempts have addressed only specific questions and no long-term live imaging data of Tribolium embryogenesis have been available. By combining light sheet-based fluorescence microscopy with a novel mounting method, we achieved complete, continuous and non-invasive fluorescence live imaging of Tribolium embryogenesis at high spatiotemporal resolution. The embryos survived the 2-day or longer imaging process, developed into adults and produced fertile progeny. Our data document all morphogenetic processes from the rearrangement of the uniform blastoderm to the onset of regular muscular movement in the same embryo and in four orientations, contributing significantly to the understanding of Tribolium development. Furthermore, we created a comprehensive chronological table of Tribolium embryogenesis, integrating most previous work and providing a reference for future studies. Based on our observations, we provide evidence that serosa window closure and serosa opening, although deferred by more than 1 day, are linked. All our long-term imaging datasets are available as a resource for the community. Tribolium is only the second insect species, after Drosophila, for which non-invasive long-term fluorescence live imaging has been achieved.

  7. Visualizing late insect embryogenesis: extraembryonic and mesodermal enhancer trap expression in the beetle Tribolium castaneum.

    PubMed

    Koelzer, Stefan; Kölsch, Yvonne; Panfilio, Kristen A

    2014-01-01

    The beetle Tribolium castaneum has increasingly become a powerful model for comparative research on insect development. One recent resource is a collection of piggyBac transposon-based enhancer trap lines. Here, we provide a detailed analysis of three selected lines and demonstrate their value for investigations in the second half of embryogenesis, which has thus far lagged behind research on early stages. Two lines, G12424 and KT650, show enhanced green fluorescent protein (EGFP) expression throughout the extraembryonic serosal tissue and in a few discrete embryonic domains. Intriguingly, both lines show for the first time a degree of regionalization within the mature serosa. However, their expression profiles illuminate distinct aspects of serosal biology: G12424 tracks the tissue's rapid maturation while KT650 expression likely reflects ongoing physiological processes. The third line, G04609, is stably expressed in mesodermal domains, including segmental muscles and the heart. Genomic mapping followed by in situ hybridization for genes near to the G04609 insertion site suggests that the transposon has trapped enhancer information for the Tribolium orthologue of midline (Tc-mid). Altogether, our analyses provide the first live imaging, long-term characterizations of enhancer traps from this collection. We show that EGFP expression is readily detected, including in heterozygote crosses that permit the simultaneous visualization of multiple tissue types. The tissue specificity provides live, endogenous marker gene expression at key developmental stages that are inaccessible for whole mount staining. Furthermore, the nonlocalized EGFP in these lines illuminates both the nucleus and cytoplasm, providing cellular resolution for morphogenesis research on processes such as dorsal closure and heart formation. In future work, identification of regulatory regions driving these enhancer traps will deepen our understanding of late developmental control, including in the

  8. Molecular and Functional Analyses of Amino Acid Decarboxylases Involved in Cuticle Tanning in Tribolium castaneum*

    PubMed Central

    Arakane, Yasuyuki; Lomakin, Joseph; Beeman, Richard W.; Muthukrishnan, Subbaratnam; Gehrke, Stevin H.; Kanost, Michael R.; Kramer, Karl J.

    2009-01-01

    Aspartate 1-decarboxylase (ADC) and 3,4-dihydroxyphenylalanine decarboxylase (DDC) provide β-alanine and dopamine used in insect cuticle tanning. β-Alanine is conjugated with dopamine to yield N-β-alanyldopamine (NBAD), a substrate for the phenol oxidase laccase that catalyzes the synthesis of cuticle protein cross-linking agents and pigment precursors. We identified ADC and DDC genes in the red flour beetle, Tribolium castaneum (Tc), and investigated their functions. TcADC mRNA was most abundant prior to the pupal-adult molt. Injection of TcADC double-stranded (ds) RNA (dsTcADC) into mature larvae resulted in depletion of NBAD in pharate adults, accumulation of dopamine, and abnormally dark pigmentation of the adult cuticle. Injection of β-alanine, the expected product of ADC, into dsTcADC-treated pupae rescued the pigmentation phenotype, resulting in normal rust-red color. A similar pattern of catechol content consisting of elevated dopamine and depressed NBAD was observed in the genetic black mutants of Tribolium, in which levels of TcADC mRNA were drastically reduced. Furthermore, from the Tribolium black mutant and dsTcADC-injected insects both exhibited similar changes in material properties. Dynamic mechanical analysis of elytral cuticle from beetles with depleted TcADC transcripts revealed diminished cross-linking of cuticular components, further confirming the important role of oxidation products of NBAD as cross-linking agents during cuticle tanning. Injection of dsTcDDC into larvae produced a lethal pupal phenotype, and the resulting grayish pupal cuticle exhibited many small patches of black pigmentation. When dsTcDDC was injected into young pupae, the resulting adults had abnormally dark brown body color, but there was little mortality. Injection of dsTcDDC resulted in more than a 5-fold increase in levels of DOPA, indicating that lack of TcDDC led to accumulation of its substrate, DOPA. PMID:19366687

  9. Anatomical and morphological spine variation in Gymnocalycium kieslingii subsp. castaneum (Cactaceae)

    PubMed Central

    Gebauer, Roman; Řepka, Radomír; Šmudla, Radek; Mamoňová, Miroslava; Ďurkovič, Jaroslav

    2016-01-01

    Abstract Although spine variation within cacti species or populations is assumed to be large, the minimum sample size of different spine anatomical and morphological traits required for species description is less studied. There are studies where only 2 spines were used for taxonomical comparison amnog species. Therefore, the spine structure variation within areoles and individuals of one population of Gymnocalycium kieslingii subsp. castaneum (Ferrari) Slaba was analyzed. Fifteen plants were selected and from each plant one areole from the basal, middle and upper part of the plant body was sampled. A scanning electron microscopy was used for spine surface description and a light microscopy for measurements of spine width, thickness, cross-section area, fiber diameter and fiber cell wall thickness. The spine surface was more visible and damaged less in the upper part of the plant body than in the basal part. Large spine and fiber differences were found between upper and lower parts of the plant body, but also within single areoles. In general, the examined traits in the upper part had by 8–17% higher values than in the lower parts. The variation of spine and fiber traits within areoles was lower than the differences between individuals. The minimum sample size was largely influenced by the studied spine and fiber traits, ranging from 1 to 70 spines. The results provide pioneer information useful in spine sample collection in the field for taxonomical, biomechanical and structural studies. Nevertheless, similar studies should be carried out for other cacti species to make generalizations. The large spine and fiber variation within areoles observed in our study indicates a very complex spine morphogenesis. PMID:27698579

  10. Visualizing Late Insect Embryogenesis: Extraembryonic and Mesodermal Enhancer Trap Expression in the Beetle Tribolium castaneum

    PubMed Central

    Koelzer, Stefan; Kölsch, Yvonne; Panfilio, Kristen A.

    2014-01-01

    The beetle Tribolium castaneum has increasingly become a powerful model for comparative research on insect development. One recent resource is a collection of piggyBac transposon-based enhancer trap lines. Here, we provide a detailed analysis of three selected lines and demonstrate their value for investigations in the second half of embryogenesis, which has thus far lagged behind research on early stages. Two lines, G12424 and KT650, show enhanced green fluorescent protein (EGFP) expression throughout the extraembryonic serosal tissue and in a few discrete embryonic domains. Intriguingly, both lines show for the first time a degree of regionalization within the mature serosa. However, their expression profiles illuminate distinct aspects of serosal biology: G12424 tracks the tissue’s rapid maturation while KT650 expression likely reflects ongoing physiological processes. The third line, G04609, is stably expressed in mesodermal domains, including segmental muscles and the heart. Genomic mapping followed by in situ hybridization for genes near to the G04609 insertion site suggests that the transposon has trapped enhancer information for the Tribolium orthologue of midline (Tc-mid). Altogether, our analyses provide the first live imaging, long-term characterizations of enhancer traps from this collection. We show that EGFP expression is readily detected, including in heterozygote crosses that permit the simultaneous visualization of multiple tissue types. The tissue specificity provides live, endogenous marker gene expression at key developmental stages that are inaccessible for whole mount staining. Furthermore, the nonlocalized EGFP in these lines illuminates both the nucleus and cytoplasm, providing cellular resolution for morphogenesis research on processes such as dorsal closure and heart formation. In future work, identification of regulatory regions driving these enhancer traps will deepen our understanding of late developmental control, including in the

  11. Expression of an endoglucanase from Tribolium castaneum (TcEG1) in Saccharomyces cerevisiae.

    PubMed

    Shirley, Derek; Oppert, Cris; Reynolds, Todd B; Miracle, Bethany; Oppert, Brenda; Klingeman, William E; Jurat-Fuentes, Juan Luis

    2014-10-01

    Insects are a largely unexploited resource in prospecting for novel cellulolytic enzymes to improve the production of ethanol fuel from lignocellulosic biomass. The cost of lignocellulosic ethanol production is expected to decrease by the combination of cellulose degradation (saccharification) and fermentation of the resulting glucose to ethanol in a single process, catalyzed by the yeast Saccharomyces cerevisiae transformed to express efficient cellulases. While S. cerevisiae is an established heterologous expression system, there are no available data on the functional expression of insect cellulolytic enzymes for this species. To address this knowledge gap, S. cerevisiae was transformed to express the full-length cDNA encoding an endoglucanase from the red flour beetle, Tribolium castaneum (TcEG1), and evaluated the activity of the transgenic product (rTcEG1). Expression of the TcEG1 cDNA in S. cerevisiae was under control of the strong glyceraldehyde-3 phosphate dehydrogenase promoter. Cultured transformed yeast secreted rTcEG1 protein as a functional β-1,4-endoglucanase, which allowed transformants to survive on selective media containing cellulose as the only available carbon source. Evaluation of substrate specificity for secreted rTcEG1 demonstrated endoglucanase activity, although some activity was also detected against complex cellulose substrates. Potentially relevant to uses in biofuel production rTcEG1 activity increased with pH conditions, with the highest activity detected at pH 12. Our results demonstrate the potential for functional production of an insect cellulase in S. cerevisiae and confirm the stability of rTcEG1 activity in strong alkaline environments.

  12. Determination of mortality of different life stages of Tribolium castaneum (Coleoptera: Tenebrionidae) in stored barley using microwaves.

    PubMed

    Vadivambal, R; Jayas, D S; White, N D G

    2008-06-01

    Barley, Hordeum vulgare L., one of the important crops in Canada, is used in malting, feed, and food industries. Disinfestation of barley using microwaves can be an alternative to chemical methods used to kill insects. A pilot-scale industrial microwave system operating at 2.45 MHz was used in this study to determine the mortality of life stages (egg, larva, pupa, and adult) of Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae). Barley samples of 50 g each at 14, 16, and 18% moisture content (MC; wet basis) were infested with various life stages of T. castaneum and exposed to microwave energy at different power levels and exposure times, and the mortality of the insects was determined. The average temperature of the 14% MC sample exposed to 28 s at 0, 200, 300, 400, and 500 W were 27.4, 42.6, 53.7, 66.9, and 73.0 degrees C, respectively, and those exposed to 56 s at 0, 200, 300, and 400 W were 27.4, 57.3, 75.5, and 91.2 degrees C, respectively. A similar range of temperature was observed for 16 and 18% MC barley. Complete mortality of all life stages of T. castaneum can be achieved at a power level of 400 W and an exposure time of 56 s or at 500 W for 28 s. Among the life stages of T. castaneum, eggs were the most susceptible to microwave energy and adults were the least susceptible. There was no significant difference in the mortality of larvae and adults at 14, 16, and 18% MC, but the mortality was significantly different for eggs and pupae at different MCs. There was a significant increase in the mortality with an increase in power level or exposure time or both. Germination capacity of the seeds was decreased with an increase in power level or exposure time or both. Quality characteristics such as alpha-amylase, diastatic power, soluble protein, viscosity, and density of the barley malt treated at 500 W for 28 s were same as the control sample, whereas the samples treated at 400 W for 56 s were significantly lower.

  13. Cloning and characterization of the Tribolium castaneum eye-color genes encoding tryptophan oxygenase and kynurenine 3-monooxygenase.

    PubMed

    Lorenzen, Marcé D; Brown, Susan J; Denell, Robin E; Beeman, Richard W

    2002-01-01

    The use of eye-color mutants and their corresponding genes as scorable marker systems has facilitated the development of transformation technology in Drosophila and other insects. In the red flour beetle, Tribolium castaneum, the only currently available system for germline transformation employs the exogenous marker gene, EGFP, driven by an eye-specific promoter. To exploit the advantages offered by eye-pigmentation markers, we decided to develop a transformant selection system for Tribolium on the basis of mutant rescue. The Tribolium orthologs of the Drosophila eye-color genes vermilion (tryptophan oxygenase) and cinnabar (kynurenine 3-monooxygenase) were cloned and characterized. Conceptual translations of Tc vermilion (Tcv) and Tc cinnabar (Tccn) are 71 and 51% identical to their respective Drosophila orthologs. We used RNA interference (RNAi) to show that T. castaneum larvae lacking functional Tcv or Tccn gene products also lack the pigmented eyespots observed in wild-type larvae. Five available eye-color mutations were tested for linkage to Tcv or Tccn via recombinational mapping. No linkage was found between candidate mutations and Tccn. However, tight linkage was found between Tcv and the white-eye mutation white, here renamed vermilion(white) (v(w)). Molecular analysis indicates that 80% of the Tcv coding region is deleted in v(w) beetles. These observations suggest that the Tribolium eye is pigmented only by ommochromes, not pteridines, and indicate that Tcv is potentially useful as a germline transformation marker. PMID:11805058

  14. Evaluation of Reference Genes for RT-qPCR in Tribolium castaneum (Coleoptera: Tenebrionidae) Under UVB Stress.

    PubMed

    Sang, Wen; He, Li; Wang, Xiao-Ping; Zhu-Salzman, Keyan; Lei, Chao-Liang

    2015-04-01

    Reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) has become a widely used technique to quantify gene expression. It is necessary to select appropriate reference genes for normalization. In the present study, we assessed the expression stability of seven candidate genes in Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae) irradiated by ultraviolet B (UVB) at different developmental stages for various irradiation time periods. The algorithms of geNorm, NormFinder, and BestKeeper were applied to determine the stability of these candidate genes. Ribosomal protein genes RpS3, RpL13A, and β-actin gene (ActB) showed the highest stability across all UVB irradiation time points, whereas expression of other normally used reference genes, such as those encoding the β-tubulin gene TUBB and the E-cadherin gene CAD, varied at different developmental stages. This study will potentially provide more suitable reference gene candidates for RT-qPCR analysis in T. castaneum subjected to environmental stresses, particularly UV irradiation.

  15. Evaluation of catmint oil and hydrogenated catmint oil as repellents for the flour beetles, Tribolium castaneum and Tribolium confusum.

    PubMed

    Arthur, Frank H; Fontenot, Emily A; Campbell, James F

    2011-01-01

    Catmint oil and hydrogenated catmint oil were evaluated as repellents for adult Tribolium casteneum (Herbst) (Coleoptera: Tenebrionidae), the red flour beetle, and T. confusum (DuVal), the confused flour beetle, using both a traditional method of visual assessment of distribution and a video recording method to determine movement patterns of individual insects. Visual assessments of distribution using groups of adults showed that the hydrogenated catmint oil was more effective than the pure catmint oil, but there was no significant difference between species. However, when repellency was measured using single insects and the visual recording system, both oils were significantly more repellent to T. castaneum than T. confusum at the concentrations evaluated in the study. Avoidance movement and change in direction when T. castaneum encountered the repellent were observed. Results indicate that repellents may be more accurately assessed using single insects rather than groups of individuals, and simple visual observations of distribution may be less sensitive in measuring repellent efficacy. Procedures for utilizing a video system are described as models for future evaluations of repellents for stored-product beetles. PMID:22235903

  16. Cloning and characterization of the Tribolium castaneum eye-color genes encoding tryptophan oxygenase and kynurenine 3-monooxygenase.

    PubMed

    Lorenzen, Marcé D; Brown, Susan J; Denell, Robin E; Beeman, Richard W

    2002-01-01

    The use of eye-color mutants and their corresponding genes as scorable marker systems has facilitated the development of transformation technology in Drosophila and other insects. In the red flour beetle, Tribolium castaneum, the only currently available system for germline transformation employs the exogenous marker gene, EGFP, driven by an eye-specific promoter. To exploit the advantages offered by eye-pigmentation markers, we decided to develop a transformant selection system for Tribolium on the basis of mutant rescue. The Tribolium orthologs of the Drosophila eye-color genes vermilion (tryptophan oxygenase) and cinnabar (kynurenine 3-monooxygenase) were cloned and characterized. Conceptual translations of Tc vermilion (Tcv) and Tc cinnabar (Tccn) are 71 and 51% identical to their respective Drosophila orthologs. We used RNA interference (RNAi) to show that T. castaneum larvae lacking functional Tcv or Tccn gene products also lack the pigmented eyespots observed in wild-type larvae. Five available eye-color mutations were tested for linkage to Tcv or Tccn via recombinational mapping. No linkage was found between candidate mutations and Tccn. However, tight linkage was found between Tcv and the white-eye mutation white, here renamed vermilion(white) (v(w)). Molecular analysis indicates that 80% of the Tcv coding region is deleted in v(w) beetles. These observations suggest that the Tribolium eye is pigmented only by ommochromes, not pteridines, and indicate that Tcv is potentially useful as a germline transformation marker.

  17. Woot, an Active Gypsy-Class Retrotransposon in the Flour Beetle, Tribolium Castaneum, Is Associated with a Recent Mutation

    PubMed Central

    Beeman, R. W.; Thomson, M. S.; Clark, J. M.; DeCamillis, M. A.; Brown, S. J.; Denell, R. E.

    1996-01-01

    A recently isolated, lethal mutation of the homeotic Abdominal gene of the red flour beetle Tribolium castaneum is associated with an insertion of a novel retrotransposon into an intron. Sequence analysis indicates that this retrotransposon, named Woot, is a member of the gypsy family of mobile elements. Most strains of T. castaneum appear to harbor ~25-35 copies of Woot per genome. Woot is composed of long terminal repeats of unprecedented length (3.6 kb each), flanking an internal coding region 5.0 kb in length. For most copies of Woot, the internal region includes two open reading frames (ORFs) that correspond to the gag and pol genes of previously described retrotransposons and retroviruses. The copy of Woot inserted into Abdominal bears an apparent single frameshift mutation that separates the normal second ORF into two. Woot does not appear to generate infectious virions by the criterion that no envelop gene is discernible. The association of Woot with a recent mutation suggests that this retroelement is currently transpositionally active in at least some strains. PMID:8722793

  18. In Vitro Comparison of the Activity Requirements and Substrate Specificity of Human and Triboleum castaneum PINK1 Orthologues

    PubMed Central

    Aerts, Liesbeth; Craessaerts, Katleen; De Strooper, Bart; Morais, Vanessa A.

    2016-01-01

    Mutations in the gene encoding the mitochondrial kinase PINK1 cause early-onset familial Parkinson’s disease. To understand the biological function of PINK1 and its role in the pathogenesis of Parkinson’s disease, it is useful to study its kinase activity towards substrates both in vivo and in vitro. For in vitro kinase assays, a purified Triboleum castaneum PINK1 insect orthologue is often employed, because it displays higher levels of activity when compared to human PINK1. We show, however, that the activity requirements, and more importantly the substrate specificity, differ between both orthologues. While Triboleum castaneum PINK1 readily phosphorylates the PINKtide peptide and Histone H1 in vitro, neither of these non-physiological substrates is phosphorylated by human PINK1. Nonetheless, both Tc and human PINK1 phosphorylate Parkin and Ubiquitin, two physiological substrates of PINK1. Our results show that the substrate selectivity differs among PINK1 orthologues, an important consideration that should be taken into account when extrapolating findings back to human PINK1. PMID:26784449

  19. Cap’n’collar differentiates the mandible from the maxilla in the beetle Tribolium castaneum

    PubMed Central

    2012-01-01

    Background The biting mandible of the arthropods is thought to have evolved in the ancestor of the insects, crustaceans and myriapods: the Mandibulata. A unique origin suggests a common set of developmental genes will be required to pattern the mandible in different arthropods. To date we have functional studies on patterning of the mandibular segment of Drosophila melanogaster showing in particular the effects of the gene cap’n’collar (cnc), however, the dipteran head is far from representative of insects or of more distantly related mandibulates; Drosophila does not even possess a mandibular appendage. To study the development of a more representative insect mandible, we chose the red flour beetle Tribolium castaneum and investigated the function of the Tribolium orthologs of cap’n’collar (Tc-cnc) and the Hox gene Deformed (Tc-Dfd). In order to determine the function of Tc-cnc and Tc-Dfd, transcripts were knocked down by maternal RNA interference (RNAi). The effects of gene knockdown were examined in the developing embryos and larvae. The effect of Tc-cnc and Tc-Dfd knockdown on the expression of other genes was determined by using in situ hybridization on Tribolium embryos. Results Our analyses show that Tc-cnc is required for specification of the identity of the mandibular segment of Tribolium and differentiates the mandible from maxillary identity. Loss of Tc-cnc function results in a transformation of the mandible to maxillary identity as well as deletion of the labrum. Tc-Dfd and the Tribolium homolog of proboscipedia (Tc-mxp = maxillopedia), Hox genes that are required to pattern the maxillary appendage, are expressed in a maxilla-like manner in the transformed mandible. Tribolium homologs of paired (Tc-prd) and Distal-less (Tc-Dll) that are expressed in the endites and telopodites of embryonic appendages are also expressed in a maxilla-like manner in the transformed mandible. We also show that Tc-Dfd is required to activate the collar of Tc

  20. Proteome Response of Tribolium castaneum Larvae to Bacillus thuringiensis Toxin Producing Strains

    PubMed Central

    Contreras, Estefanía; Rausell, Carolina; Real, M. Dolores

    2013-01-01

    Susceptibility of Tribolium castaneum (Tc) larvae was determined against spore-crystal mixtures of five coleopteran specific and one lepidopteran specific Bacillus thuringiensis Cry toxin producing strains and those containing the structurally unrelated Cry3Ba and Cry23Aa/Cry37Aa proteins were found toxic (LC50 values 13.53 and 6.30 µg spore-crystal mixture/µL flour disc, respectively). Using iTRAQ combined with LC-MS/MS allowed the discovery of seven novel differentially expressed proteins in early response of Tc larvae to the two active spore-crystal mixtures. Proteins showing a statistically significant change in treated larvae compared to non-intoxicated larvae fell into two major categories; up-regulated proteins were involved in host defense (odorant binding protein C12, apolipophorin-III and chemosensory protein 18) and down-regulated proteins were linked to metabolic pathways affecting larval metabolism and development (pyruvate dehydrogenase Eα subunit, cuticular protein, ribosomal protein L13a and apolipoprotein LI-II). Among increased proteins, Odorant binding protein C12 showed the highest change, 4-fold increase in both toxin treatments. The protein displayed amino acid sequence and structural homology to Tenebrio molitor 12 kDa hemolymph protein b precursor, a non-olfactory odorant binding protein. Analysis of mRNA expression and mortality assays in Odorant binding protein C12 silenced larvae were consistent with a general immune defense function of non-olfactory odorant binding proteins. Regarding down-regulated proteins, at the transcriptional level, pyruvate dehydrogenase and cuticular genes were decreased in Tc larvae exposed to the Cry3Ba producing strain compared to the Cry23Aa/Cry37Aa producing strain, which may contribute to the developmental arrest that we observed with larvae fed the Cry3Ba producing strain. Results demonstrated a distinct host transcriptional regulation depending upon the Cry toxin treatment. Knowledge on how insects

  1. Proteome response of Tribolium castaneum larvae to Bacillus thuringiensis toxin producing strains.

    PubMed

    Contreras, Estefanía; Rausell, Carolina; Real, M Dolores

    2013-01-01

    Susceptibility of Tribolium castaneum (Tc) larvae was determined against spore-crystal mixtures of five coleopteran specific and one lepidopteran specific Bacillus thuringiensis Cry toxin producing strains and those containing the structurally unrelated Cry3Ba and Cry23Aa/Cry37Aa proteins were found toxic (LC(50) values 13.53 and 6.30 µg spore-crystal mixture/µL flour disc, respectively). Using iTRAQ combined with LC-MS/MS allowed the discovery of seven novel differentially expressed proteins in early response of Tc larvae to the two active spore-crystal mixtures. Proteins showing a statistically significant change in treated larvae compared to non-intoxicated larvae fell into two major categories; up-regulated proteins were involved in host defense (odorant binding protein C12, apolipophorin-III and chemosensory protein 18) and down-regulated proteins were linked to metabolic pathways affecting larval metabolism and development (pyruvate dehydrogenase Eα subunit, cuticular protein, ribosomal protein L13a and apolipoprotein LI-II). Among increased proteins, Odorant binding protein C12 showed the highest change, 4-fold increase in both toxin treatments. The protein displayed amino acid sequence and structural homology to Tenebrio molitor 12 kDa hemolymph protein b precursor, a non-olfactory odorant binding protein. Analysis of mRNA expression and mortality assays in Odorant binding protein C12 silenced larvae were consistent with a general immune defense function of non-olfactory odorant binding proteins. Regarding down-regulated proteins, at the transcriptional level, pyruvate dehydrogenase and cuticular genes were decreased in Tc larvae exposed to the Cry3Ba producing strain compared to the Cry23Aa/Cry37Aa producing strain, which may contribute to the developmental arrest that we observed with larvae fed the Cry3Ba producing strain. Results demonstrated a distinct host transcriptional regulation depending upon the Cry toxin treatment. Knowledge on how insects

  2. Evaluation of Standard Loose Plastic Packaging for the Management of Rhyzopertha dominica (F.) (Coleoptera: Bostrichidae) and Tribolium castaneum (Herbst) (Coleoptera: Tenebriondiae).

    PubMed

    Hassan, Muhammad Waqar; Gulraize; Ali, Usman; Ur Rehman, Fazal; Najeeb, Hafsa; Sohail, Maryam; Irsa, Bakhtawar; Muzaffar, Zubaria; Chaudhry, Muhammad Shafiq

    2016-01-01

    Three standard foodstuff plastic packaging namely polyethylene (PE), polypropylene (PP), and polyvinylchloride (PVC) were evaluated for management of lesser grain borer Rhyzopertha dominica (F.) (Coleoptera: Bostrichidae) and red flour beetle Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae). Resistance parameters in packaging were recorded as punctures, holes, penetrations, sealing defects, and invasions with two thicknesses and tested for two lengths of time. Damages like punctures, holes and penetrations by both insects were more in PE packaging however R. dominica made more penetrations in PP than in PE. For both insects sealing defects and invasions were predominant in PVC than in others. Thickness did not affect significantly damage types but significantly more holes and penetrations by R. dominica were in less thickness. Punctures and holes by R. dominica were more after less time period but other damages in packaging were more after more time period. However for T. castaneum all sorts of damages were seen more after more time period. Overall categorization between two insects showed R. dominica made more penetrations and T. castaneum made more invasions compared with their counterparts. Pictures were taken under camera fitted microscope to magnify punctures and holes in different packaging and thicknesses. Insect mortality due to phosphine was more in PP and PE packaging and least in PVC packaging and thickness effect was marginal. T. castaneum mortality was significantly more after 48 h than after 24 h. Damages extent in packaging and fumigation results showed PP to be the best of three packaging materials to manage these insects.

  3. Genes encoding proteins with peritrophin A-type chitin-binding domains in Tribolium castaneum are grouped into three distinct families based on phylogeny, expression and function

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This study is focused on the characterization and expression of genes in the red flour beetle, Tribolium castaneum, encoding proteins that possess six-cysteine-containing chitin-binding domains (CBDs) related to the peritrophin A domain (ChtBD2). An exhaustive bioinformatics search of the genome of...

  4. Evaluation of Standard Loose Plastic Packaging for the Management of Rhyzopertha dominica (F.) (Coleoptera: Bostrichidae) and Tribolium castaneum (Herbst) (Coleoptera: Tenebriondiae)

    PubMed Central

    Hassan, Muhammad Waqar; Gulraize; Ali, Usman; Ur Rehman, Fazal; Najeeb, Hafsa; Sohail, Maryam; Irsa, Bakhtawar; Muzaffar, Zubaria; Chaudhry, Muhammad Shafiq

    2016-01-01

    Three standard foodstuff plastic packaging namely polyethylene (PE), polypropylene (PP), and polyvinylchloride (PVC) were evaluated for management of lesser grain borer Rhyzopertha dominica (F.) (Coleoptera: Bostrichidae) and red flour beetle Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae). Resistance parameters in packaging were recorded as punctures, holes, penetrations, sealing defects, and invasions with two thicknesses and tested for two lengths of time. Damages like punctures, holes and penetrations by both insects were more in PE packaging however R. dominica made more penetrations in PP than in PE. For both insects sealing defects and invasions were predominant in PVC than in others. Thickness did not affect significantly damage types but significantly more holes and penetrations by R. dominica were in less thickness. Punctures and holes by R. dominica were more after less time period but other damages in packaging were more after more time period. However for T. castaneum all sorts of damages were seen more after more time period. Overall categorization between two insects showed R. dominica made more penetrations and T. castaneum made more invasions compared with their counterparts. Pictures were taken under camera fitted microscope to magnify punctures and holes in different packaging and thicknesses. Insect mortality due to phosphine was more in PP and PE packaging and least in PVC packaging and thickness effect was marginal. T. castaneum mortality was significantly more after 48 h than after 24 h. Damages extent in packaging and fumigation results showed PP to be the best of three packaging materials to manage these insects. PMID:27638958

  5. Residual efficacy of methoprene for control of Tribolium castaneum (Coleoptera: Tenebrionidae) larvae at different temperatures on varnished wood, concrete, and wheat

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The residual efficacy of the juvenile hormone analogue, methoprene (Diacon II), was evaluated in bioassays using larvae of Tribolium castaneum (Herbst) exposed on varnished wood or unsealed concrete treated with a liquid formulation and held at different temperatures. When these surfaces were stored...

  6. Evaluation of structural treatment efficacy against Tribolium castaneum and Tribolium confusum (Coleoptera: Tenebrionidae) using meta-analysis of multiple studies conducted in food facilities

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The phase out of methyl bromide for the treatment of structures where grain is milled and processed has triggered a need to evaluate the effectiveness of alternative structural treatments such as sulfuryl fluoride and heat. The red flour beetle (Tribolium castaneum (Herbst)) (RFB) and confused flour...

  7. Impact of varying levels of sanitation on mortality of Tribolium castaneum eggs and adults during heat treatment of a pilot flour mill

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The influence of sanitation on responses of life stages of the red flour beetle, Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae), an economically important pest in flour mills, was investigated in a pilot flour mill subjected to two, 24-h heat treatments. One hundred eggs or 100 adults of T...

  8. Evaluation of Standard Loose Plastic Packaging for the Management of Rhyzopertha dominica (F.) (Coleoptera: Bostrichidae) and Tribolium castaneum (Herbst) (Coleoptera: Tenebriondiae).

    PubMed

    Hassan, Muhammad Waqar; Gulraize; Ali, Usman; Ur Rehman, Fazal; Najeeb, Hafsa; Sohail, Maryam; Irsa, Bakhtawar; Muzaffar, Zubaria; Chaudhry, Muhammad Shafiq

    2016-01-01

    Three standard foodstuff plastic packaging namely polyethylene (PE), polypropylene (PP), and polyvinylchloride (PVC) were evaluated for management of lesser grain borer Rhyzopertha dominica (F.) (Coleoptera: Bostrichidae) and red flour beetle Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae). Resistance parameters in packaging were recorded as punctures, holes, penetrations, sealing defects, and invasions with two thicknesses and tested for two lengths of time. Damages like punctures, holes and penetrations by both insects were more in PE packaging however R. dominica made more penetrations in PP than in PE. For both insects sealing defects and invasions were predominant in PVC than in others. Thickness did not affect significantly damage types but significantly more holes and penetrations by R. dominica were in less thickness. Punctures and holes by R. dominica were more after less time period but other damages in packaging were more after more time period. However for T. castaneum all sorts of damages were seen more after more time period. Overall categorization between two insects showed R. dominica made more penetrations and T. castaneum made more invasions compared with their counterparts. Pictures were taken under camera fitted microscope to magnify punctures and holes in different packaging and thicknesses. Insect mortality due to phosphine was more in PP and PE packaging and least in PVC packaging and thickness effect was marginal. T. castaneum mortality was significantly more after 48 h than after 24 h. Damages extent in packaging and fumigation results showed PP to be the best of three packaging materials to manage these insects. PMID:27638958

  9. Loss of function of the yellow-e gene causes dehydration-induced mortality of adult Tribolium castaneum.

    PubMed

    Noh, Mi Young; Kramer, Karl J; Muthukrishnan, Subbaratnam; Beeman, Richard W; Kanost, Michael R; Arakane, Yasuyuki

    2015-03-15

    Yellow protein (dopachrome conversion enzyme, DCE) is involved in the melanin biosynthetic pathway that significantly accelerates pigmentation reactions in insects. Recent studies have suggested that the insect yellow genes represent a rapidly evolving gene family generating functionally diverse paralogs, but the exact physiological functions of several yellow genes are still not understood. To study the function(s) of one of the yellow genes, yellow-e (TcY-e), in the red flour beetle, Tribolium castaneum, we performed real-time PCR to analyze its developmental and tissue-specific expression, and utilized immunohistochemistry to identify the localization of the TcY-e protein in adult cuticle. Injection of double-stranded RNA for TcY-e (dsTcY-e) into late instar larvae had no effect on larval-pupal molting or pupal development. The pupal cuticle, including that lining the setae, gin traps and urogomphi, underwent normal tanning. Adult cuticle tanning including that of the head, mandibles and legs viewed through the translucent pupal cuticle was initiated on schedule (pupal days 4-5), indicating that TcY-e is not required for pupal or pharate adult cuticle pigmentation in T. castaneum. The subsequent pupal-adult molt, however, was adversely affected. Although pupal cuticle apolysis and slippage were evident, some of the adults (~25%) were unable to shed their exuvium and died entrapped in their pupal cuticle. In addition, the resulting adults rapidly became highly desiccated. Interestingly, both the failure of the pupal-adult molt and desiccation-induced mortality were prevented by maintaining the dsTcY-e-treated insects at 100% relative humidity (rh). However, when the high humidity-rescued adults were removed from 100% rh and transferred to 50% rh, they rapidly dehydrated and died, whereas untreated beetles thrived throughout development at 50% rh. We also observed that the body color of the high humidity-rescued dsTcY-e-adults was slightly darker than that of

  10. Knickkopf and retroactive proteins are required for formation of laminar serosal procuticle during embryonic development of Tribolium castaneum.

    PubMed

    Chaudhari, Sujata S; Noh, Mi Young; Moussian, Bernard; Specht, Charles A; Kramer, Karl J; Beeman, Richard W; Arakane, Yasuyuki; Muthukrishnan, Subbaratnam

    2015-05-01

    Chitin, a homopolymer of β-1-4-linked N-acetylglucosamine synthesized by chitin synthase A (Chs-A), is organized in the procuticle of the postembryonic cuticle or exoskeleton, which is composed of laminae stacked parallel to the cell surface to give stability and integrity to the underlying insect epidermal and other tissues. Our previous work has revealed an important role for two proteins from Tribolium castaneum named Knickkopf (TcKnk) and Retroactive (TcRtv) in postembryonic cuticular chitin maintenance. TcKnk and TcRtv were shown to be required for protection and organization of newly synthesized procuticular chitin. To study the functions of TcKnk and TcRtv in serosal and larval cuticles produced during embryogenesis in T. castaneum, dsRNAs specific for these two genes were injected into two week-old adult females. The effects of dsRNA treatment on ovarial integrity, oviposition, egg hatching and adult survival were determined. Insects treated with dsRNA for chitin synthase-A (TcChs-A) and tryptophan oxygenase (TcVer) were used as positive and negative controls for these experiments, respectively. Like TcChs-A RNAi, injection of dsRNA for TcKnk or TcRtv into adult females exhibited no adult lethality and oviposition was normal. However, a vast majority of the embryos did not hatch. The remaining (∼10%) of the embryos hatched into first instar larvae that died without molting to the second instar. Chitin content analysis following TcKnk and TcRtv parental RNAi revealed approximately 50% reduction in chitin content of eggs in comparison with control TcVer RNAi, whereas TcChs-A dsRNA-treatment led to >90% loss of chitin. Furthermore, transmission electron microscopic (TEM) analysis of serosal cuticle from TcChs-A, TcKnk and TcRtv dsRNA-treated insects revealed a complete absence of laminar organization of serosal (and larval) procuticle in comparison with TcVer dsRNA-treated controls, which exhibited normal laminar organization of procuticular chitin. The

  11. Translocation of bacteria from the gut to the eggs triggers maternal transgenerational immune priming in Tribolium castaneum

    PubMed Central

    Knorr, Eileen; Schmidtberg, Henrike; Arslan, Derya; Bingsohn, Linda; Vilcinskas, Andreas

    2015-01-01

    Invertebrates can be primed to enhance their protection against pathogens they have encountered before. This enhanced immunity can be passed maternally or paternally to the offspring and is known as transgenerational immune priming. We challenged larvae of the red flour beetle Tribolium castaneum by feeding them on diets supplemented with Escherichia coli, Micrococcus luteus or Pseudomonas entomophila, thus mimicking natural exposure to pathogens. The oral uptake of bacteria induced immunity-related genes in the offspring, but did not affect the methylation status of the egg DNA. However, we observed the translocation of bacteria or bacterial fragments from the gut to the developing eggs via the female reproductive system. Such translocating microbial elicitors are postulated to trigger bacterial strain-specific immune responses in the offspring and provide an alternative mechanistic explanation for maternal transgenerational immune priming in coleopteran insects. PMID:26701756

  12. A Defensin from the Model Beetle Tribolium castaneum Acts Synergistically with Telavancin and Daptomycin against Multidrug Resistant Staphylococcus aureus.

    PubMed

    Rajamuthiah, Rajmohan; Jayamani, Elamparithi; Conery, Annie L; Fuchs, Beth Burgwyn; Kim, Wooseong; Johnston, Tatiana; Vilcinskas, Andreas; Ausubel, Frederick M; Mylonakis, Eleftherios

    2015-01-01

    The red flour beetle Tribolium castaneum is a common insect pest and has been established as a model beetle to study insect development and immunity. This study demonstrates that defensin 1 from T. castaneum displays in vitro and in vivo antimicrobial activity against drug resistant Staphylococcus aureus strains. The minimum inhibitory concentration (MIC) of defensin 1 against 11 reference and clinical staphylococcal isolates was between 16-64 μg/ml. The putative mode of action of the defensin peptide is disruption of the bacterial cell membrane. The antibacterial activity of defensin 1 was attenuated by salt concentrations of 1.56 mM and 25 mM for NaCl and CaCl2 respectively. Treatment of defensin 1 with the reducing agent dithiothreitol (DTT) at concentrations 1.56 to 3.13 mM abolished the antimicrobial activity of the peptide. In the presence of subinhibitory concentrations of antibiotics that also target the bacterial cell envelope such as telavancin and daptomycin, the MIC of the peptide was as low as 1 μg/ml. Moreover, when tested against an S. aureus strain that was defective in D-alanylation of the cell wall, the MIC of the peptide was 0.5 μg/ml. Defensin 1 exhibited no toxicity against human erythrocytes even at 400 μg/ml. The in vivo activity of the peptide was validated in a Caenorhabditis elegans-MRSA liquid infection assay. These results suggest that defensin 1 behaves similarly to other cationic AMPs in its mode of action against S. aureus and that the activity of the peptide can be enhanced in combination with other antibiotics with similar modes of action or with compounds that have the ability to decrease D-alanylation of the bacterial cell wall.

  13. Transcription profiling of immune genes during parasite infection in susceptible and resistant strains of the flour beetles (Tribolium castaneum).

    PubMed

    Zhong, Daibin; Wang, Mei-Hui; Pai, Aditi; Yan, Guiyun

    2013-05-01

    The flour beetle, Tribolium castaneum, is an intermediate host for the tapeworm Hymenolepis diminuta and has become an important genetic model to explore immune responses to parasite infection in insect hosts. The present study examined the immune responses to tapeworm infection in resistant (TIW1) and susceptible (cSM) strains of the red flour beetle, T. castaneum, using real-time quantitative reverse transcription PCR on 29 immunity-related genes that exhibit antimicrobial properties. Thirteen of the 29 genes showed constitutive differences in expression between the two strains. Fourteen to fifteen of the 29 genes exhibited significant differences in transcription levels when beetles were challenged with tapeworm parasite in the resistant and susceptible strains. Nine genes (GNBP3, cSPH2, lysozyme4, defensin1, PGRP-SA, defensin2, coleoptericin1, attacin2 and serpin29) in cSM and 13 genes (lysozyme2, proPO1, GNBP3, cSPH2, lysozyme4, defensin1, PGRP-SA, defensin2, coleoptericin1, attacin2, proPO2/3, PGRP-LE and PGRP-SB) in TIW1 were up-regulated by infections or showed parasite infection-induced expression. Seven genes (attacin2, coleoptericin1, defensin1, defensin2, lysozyme2, PGRP-SA and PGRP-SB) were more than 10 folds higher in the resistant TIW1 strain than in the susceptible cSM strain after exposure to tapeworm parasites. This study demonstrated the effects of genetic background, the transcription profile to parasite infection, and identified the immunity-related genes that were significantly regulated by the infection of tapeworms in Tribolium beetles.

  14. Tribolium castaneum RR-1 Cuticular Protein TcCPR4 Is Required for Formation of Pore Canals in Rigid Cuticle

    PubMed Central

    Noh, Mi Young; Muthukrishnan, Subbaratnam; Kramer, Karl J.; Arakane, Yasuyuki

    2015-01-01

    Insect cuticle is composed mainly of structural proteins and the polysaccharide chitin. The CPR family is the largest family of cuticle proteins (CPs), which can be further divided into three subgroups based on the presence of one of the three presumptive chitin-binding sequence motifs denoted as Rebers-Riddiford (R&R) consensus sequence motifs RR-1, RR-2 and RR-3. The TcCPR27 protein containing the RR-2 motif is one of the most abundant CPs present both in the horizontal laminae and in vertical pore canals in the procuticle of rigid cuticle found in the elytron of the red flour beetle, Tribolium castaneum. Depletion of TcCPR27 by RNA interference (RNAi) causes both unorganized laminae and pore canals, resulting in malformation and weakening of the elytron. In this study, we investigated the function(s) of another CP, TcCPR4, which contains the RR-1 motif and is easily extractable from elytra after RNAi to deplete the level of TcCPR27. Transcript levels of the TcCPR4 gene are dramatically increased in 3 d-old pupae when adult cuticle synthesis begins. Immunohistochemical studies revealed that TcCPR4 protein is present in the rigid cuticles of the dorsal elytron, ventral abdomen and leg but not in the flexible cuticles of the hindwing and dorsal abdomen of adult T. castaneum. Immunogold labeling and transmission electron microscopic analyses revealed that TcCPR4 is predominantly localized in pore canals and regions around the apical plasma membrane protrusions into the procuticle of rigid adult cuticles. RNAi for TcCPR4 resulted in an abnormal shape of the pore canals with amorphous pore canal fibers (PCFs) in their lumen. These results support the hypothesis that TcCPR4 is required for achieving proper morphology of the vertical pore canals and PCFs that contribute to the assembly of a cuticle that is both lightweight and rigid. PMID:25664770

  15. A Defensin from the Model Beetle Tribolium castaneum Acts Synergistically with Telavancin and Daptomycin against Multidrug Resistant Staphylococcus aureus

    PubMed Central

    Rajamuthiah, Rajmohan; Jayamani, Elamparithi; Conery, Annie L.; Fuchs, Beth Burgwyn; Kim, Wooseong; Johnston, Tatiana; Vilcinskas, Andreas; Ausubel, Frederick M.; Mylonakis, Eleftherios

    2015-01-01

    The red flour beetle Tribolium castaneum is a common insect pest and has been established as a model beetle to study insect development and immunity. This study demonstrates that defensin 1 from T. castaneum displays in vitro and in vivo antimicrobial activity against drug resistant Staphylococcus aureus strains. The minimum inhibitory concentration (MIC) of defensin 1 against 11 reference and clinical staphylococcal isolates was between 16–64 μg/ml. The putative mode of action of the defensin peptide is disruption of the bacterial cell membrane. The antibacterial activity of defensin 1 was attenuated by salt concentrations of 1.56 mM and 25 mM for NaCl and CaCl2 respectively. Treatment of defensin 1 with the reducing agent dithiothreitol (DTT) at concentrations 1.56 to 3.13 mM abolished the antimicrobial activity of the peptide. In the presence of subinhibitory concentrations of antibiotics that also target the bacterial cell envelope such as telavancin and daptomycin, the MIC of the peptide was as low as 1 μg/ml. Moreover, when tested against an S. aureus strain that was defective in D-alanylation of the cell wall, the MIC of the peptide was 0.5 μg/ml. Defensin 1 exhibited no toxicity against human erythrocytes even at 400 μg/ml. The in vivo activity of the peptide was validated in a Caenorhabditis elegans-MRSA liquid infection assay. These results suggest that defensin 1 behaves similarly to other cationic AMPs in its mode of action against S. aureus and that the activity of the peptide can be enhanced in combination with other antibiotics with similar modes of action or with compounds that have the ability to decrease D-alanylation of the bacterial cell wall. PMID:26062137

  16. Impact of varying levels of sanitation on mortality of Tribolium castaneum eggs and adults during heat treatment of a pilot flour mill.

    PubMed

    Brijwani, Monika; Subramanyam, Bhadriraju; Flinn, Paul W

    2012-04-01

    The influence of sanitation on responses of life stages of the red flour beetle, Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae), an economically important pest in flour mills, was investigated in a pilot flour mill subjected to two, 24-h heat treatments. One hundred eggs or 100 adults of T. castaneum were exposed inside each 20-cm diameter by 15-cm high PVC rings holding 0.1-, 0.2-, 1.0-, 3.0-, 6.0-, and 10.0-cm-deep wheat flour to simulate different sanitation levels that may exist in a flour mill. These rings were placed on the first and third floors of a pilot flour mill. On the first floor, temperatures inside rings with eggs reached 50 degrees C in 7-11 h only in 0.1- and 0.2-cm-deep flour treatments. In all other treatments the maximum temperatures attained generally were below 50 degrees C and inversely related to flour depth. Adults of T. castaneum on this floor were less susceptible than eggs. The egg mortality decreased linearly with an increase in flour depth, whereas that of adults decreased exponentially. All eggs and adults in rings on the third floor were killed irrespective of flour depth, because temperatures inside rings reached 50 degrees C in 15-17 h and were held above 50 degrees C for 6-8 h with the maximum temperatures ranging between 55.0 and 57.0 degrees C. Although the protective effects of flour on survival of T. castaneum eggs and adults were evident only if temperatures did not reach 50 degrees C, removal of flour accumulations is essential to improve heat treatment effectiveness.

  17. Mortality and testicular derangements in red flour beetles, Tribolium castaneum (Herbst) exposed to hen's egg white proteins.

    PubMed

    Parshad, Ranjit K; Kansal, Megha

    2012-03-01

    Red flour beetle (T. castaneum) is a major pest of stored grains and is known for its adaptability to all classes of insecticides. The present study was carried out to determine the insecticidal potential of egg white proteins to manage beetle population. Protein samples obtained through salt fractionation were lyophilized and were used separately and simultaneously in different concentrations by adding them to wheat flour and milk powder. The results indicated that the mortality rate of the adult beetles was dependent on the type of treatment, concentration of protein samples and duration of feeding. In multiple-choice feeding trials beetles showed their movement towards the control section as the concentration of treatment increases. Marked abnormalities were observed in appearance and dimensions of the testes which indicated that the egg white proteins caused considerable effect on the process of spermatogenesis and sperm functions. SEM study revealed the formation of deep wrinkles and folds on the testicular surface of the testes of beetles fed on treated diets, points towards the depletion of internal cellular material. The results suggest that egg white protein affects the survival and cause subsequent derangements in the testis of red flour beetle. PMID:22439439

  18. RNAi-mediated knockdown of the voltage gated sodium ion channel TcNav causes mortality in Tribolium castaneum.

    PubMed

    Abd El Halim, Hesham M; Alshukri, Baida M H; Ahmad, Munawar S; Nakasu, Erich Y T; Awwad, Mohammed H; Salama, Elham M; Gatehouse, Angharad M R; Edwards, Martin G

    2016-01-01

    The voltage-gated sodium ion channel (VGSC) belongs to the largest superfamily of ion channels. Since VGSCs play key roles in physiological processes they are major targets for effective insecticides. RNA interference (RNAi) is widely used to analyse gene function, but recently, it has shown potential to contribute to novel strategies for selectively controlling agricultural insect pests. The current study evaluates the delivery of dsRNA targeted to the sodium ion channel paralytic A (TcNav) gene in Tribolium castaneum as a viable means of controlling this insect pest. Delivery of TcNav dsRNA caused severe developmental arrest with larval mortalities up to 73% post injection of dsRNA. Injected larvae showed significant (p < 0.05) knockdown in gene expression between 30-60%. Expression was also significantly (p < 0.05) reduced in pupae following injection causing 30% and 42% knockdown for early and late pupal stages, respectively. Oral delivery of dsRNA caused dose-dependant mortalities of between 19 and 51.34%; this was accompanied by significant (p < 0.05) knockdown in gene expression following 3 days of continuous feeding. The majority of larvae injected with, or fed, dsRNA died during the final larval stage prior to pupation. This work provides evidence of a viable RNAi-based strategy for insect control. PMID:27411529

  19. A Soluble Pyrophosphatase Is Essential to Oogenesis and Is Required for Polyphosphate Metabolism in the Red Flour Beetle (Tribolium castaneum)

    PubMed Central

    Carvalho, Klébea; Ribeiro, Lupis; Moraes, Jorge; da Silva, José Roberto; Costa, Evenilton P.; Souza-Menezes, Jackson; Logullo, Carlos; Nunes da Fonseca, Rodrigo; Campos, Eldo

    2015-01-01

    Polyphosphates have been found in all cell types examined to date and play diverse roles depending on the cell type. In eukaryotic organisms, polyphosphates have been mainly investigated in mammalian cells with few studies on insects. Some studies have demonstrated that a pyrophosphatase regulates polyphosphate metabolism, and most of them were performed on trypanosomatids. Here, we investigated the effects of sPPase gene knocked down in oogenesis and polyphosphate metabolism in the red flour beetle (Tribolium castaneum) A single sPPase gene was identified in insect genome and is maternally provided at the mRNA level and not restricted to any embryonic or extraembryonic region during embryogenesis. After injection of Tc-sPPase dsRNA, female survival was reduced to 15% of the control (dsNeo RNA), and egg laying was completely impaired. The morphological analysis by nuclear DAPI staining of the ovarioles in Tc-sPPase dsRNA-injected females showed that the ovariole number is diminished, degenerated oocytes can be observed, and germarium is reduced. The polyphosphate level was increased in cytoplasmic and nuclear fractions in Tc-sPPase RNAi; Concomitantly, the exopolyphosphatase activity decreased in both fractions. Altogether, these data suggest a role for sPPase in the regulation on polyphosphate metabolism in insects and provide evidence that Tc-sPPase is essential to oogenesis. PMID:25811926

  20. Protein N-glycosylation and N-glycan trimming are required for postembryonic development of the pest beetle Tribolium castaneum

    PubMed Central

    Walski, Tomasz; Van Damme, Els J. M.; Smargiasso, Nicolas; Christiaens, Olivier; De Pauw, Edwin; Smagghe, Guy

    2016-01-01

    In holometabolous insects the transition from larva to adult requires a complete body reorganization and relies on N-glycosylated proteins. N-glycosylation is an important posttranslational modification that influences protein activity but its impact on the metamorphosis has not been studied yet. Here we used the red flour beetle, Tribolium castaneum, to perform a first comprehensive study on the involvement of the protein N-glycosylation pathway in metamorphosis. The transcript levels for genes encoding N-glycan processing enzymes increased during later developmental stages and, in turn, transition from larva to adult coincided with an enrichment of more extensively modified paucimannose glycans, including fucosylated ones. Blockage of N-glycan attachment resulted in larval mortality, while RNAi of α-glucosidases involved in early N-glycan trimming and quality control disrupted the larva to pupa transition. Additionally, simultaneous knockdown of multiple genes responsible for N-glycan processing towards paucimannose structures revealed their novel roles in pupal appendage formation and adult eclosion. Our findings revealed that, next to hormonal control, insect post-embryonic development and metamorphosis depend on protein N-glycan attachment and efficient N-glycan processing. Consequently, disruption of these processes could be an effective new approach for insect control. PMID:27731363

  1. The role of epigenetics in host-parasite coevolution: lessons from the model host insects Galleria mellonella and Tribolium castaneum.

    PubMed

    Vilcinskas, Andreas

    2016-08-01

    Recent studies addressing experimental host-parasite coevolution and transgenerational immune priming in insects provide evidence for heritable shifts in host resistance or parasite virulence. These rapid reciprocal adaptations may thus be transferred to offspring generations by either genetic changes or mechanisms that do not involve changes in the germline DNA sequence. Epigenetic inheritance refers to changes in gene expression that are heritable across generations and mediated by epigenetic modifications passed from parents to offspring. Highlighting the role of epigenetics in host-parasite coevolution, this review discusses the involvement of DNA methylation, histone acetylation/deacetylation and microRNAs in the interactions between bacterial or fungal parasites and model host insects such as the greater wax moth Galleria mellonella and the red flour beetle Tribolium castaneum. These epigenetic mechanisms are thought to participate in generation-spanning transcriptional reprogramming in the host insect, often linking immunity with developmentally related gene expression and contributing to the heredity of acquired adaptations. It is proposed that the interactions during host-parasite coevolution can therefore be expanded beyond reciprocal genetic changes to include reciprocal epigenetic changes. Epigenetics is thus a promising and prospering field in the context of host-parasite coevolution. PMID:27341739

  2. Differences in Attack Avoidance and Mating Success between Strains Artificially Selected for Dispersal Distance in Tribolium castaneum.

    PubMed

    Matsumura, Kentarou; Miyatake, Takahisa

    2015-01-01

    Individuals of both dispersal and non-dispersal types (disperser and non-disperser) are found in a population, suggesting that each type has both costs and benefits for fitness. However, few studies have examined the trade-off between the costs and benefits for the types. Here, we artificially selected for walking distance, i.e., an indicator of dispersal ability, in the red flour beetle Tribolium castaneum and established strains with longer (L-strains) or shorter (S-strains) walking distances. We then compared the frequency of predation by the assassin bug Amphibolus venator and the mating frequency of the selected strains. L-strain beetles suffered higher predation risk, than did S-strain beetles. L-strain males had significantly increased mating success compared to S-strain males, but females did not show a significant difference between the strains. The current results showed the existence of a trade-off between predation avoidance and mating success associated with dispersal types at a genetic level only in males. This finding can help to explain the maintenance of variation in dispersal ability within a population. PMID:25970585

  3. RNAi-mediated knockdown of the voltage gated sodium ion channel TcNav causes mortality in Tribolium castaneum

    PubMed Central

    Abd El Halim, Hesham M.; Alshukri, Baida M. H.; Ahmad, Munawar S.; Nakasu, Erich Y. T.; Awwad, Mohammed H.; Salama, Elham M.; Gatehouse, Angharad M. R.; Edwards, Martin G.

    2016-01-01

    The voltage-gated sodium ion channel (VGSC) belongs to the largest superfamily of ion channels. Since VGSCs play key roles in physiological processes they are major targets for effective insecticides. RNA interference (RNAi) is widely used to analyse gene function, but recently, it has shown potential to contribute to novel strategies for selectively controlling agricultural insect pests. The current study evaluates the delivery of dsRNA targeted to the sodium ion channel paralytic A (TcNav) gene in Tribolium castaneum as a viable means of controlling this insect pest. Delivery of TcNav dsRNA caused severe developmental arrest with larval mortalities up to 73% post injection of dsRNA. Injected larvae showed significant (p < 0.05) knockdown in gene expression between 30–60%. Expression was also significantly (p < 0.05) reduced in pupae following injection causing 30% and 42% knockdown for early and late pupal stages, respectively. Oral delivery of dsRNA caused dose-dependant mortalities of between 19 and 51.34%; this was accompanied by significant (p < 0.05) knockdown in gene expression following 3 days of continuous feeding. The majority of larvae injected with, or fed, dsRNA died during the final larval stage prior to pupation. This work provides evidence of a viable RNAi-based strategy for insect control. PMID:27411529

  4. The ABCs of Eye Color in Tribolium castaneum: Orthologs of the Drosophila white, scarlet, and brown Genes

    PubMed Central

    Grubbs, Nathaniel; Haas, Sue; Beeman, Richard W.; Lorenzen, Marcé D.

    2015-01-01

    In Drosophila melanogaster, each of the three paralogous ABC transporters, White, Scarlet and Brown, is required for normal pigmentation of the compound eye. We have cloned the three orthologous genes from the beetle Tribolium castaneum. Conceptual translations of Tribolium white (Tcw), scarlet (Tcst), and brown (Tcbw) are 51, 48, and 32% identical to their respective Drosophila counterparts. We have identified loss-of-eye-pigment strains that bear mutations in Tcw and Tcst: the Tcw gene in the ivory (i) strain carries a single-base transversion, which leads to an E → D amino-acid substitution in the highly conserved Walker B motif, while the Tcst gene in the pearl (p) strain has a deletion resulting in incorporation of a premature stop codon. In light of these findings, the mutant strains i and p are herein renamed whiteivory (wi) and scarletpearl (stp), respectively. In addition, RNA inhibition of Tcw and Tcst recapitulates the mutant phenotypes, confirming the roles of these genes in normal eye pigmentation, while RNA interference of Tcbw provides further evidence that it has no role in eye pigmentation in Tribolium. We also consider the evolutionary implications of our findings. PMID:25555987

  5. Effects of Flour and Milling Debris on Efficacy of Beta-Cyfluthrin for Control of Tribolium castaneum (Coleoptera: Tenebrionidae).

    PubMed

    Arthur, Frank H; Starkus, Laura A; Mckay, Tanja

    2015-04-01

    Laboratory and field studies were conducted to determine if accumulation of a flour food source or milling debris affected residual efficacy of beta-cyfluthrin for control of Tribolium castaneum (Herbst), the red flour beetle. In the laboratory study, the high label rate of 20 mg active ingredient (AI)/m2 gave effective control for 8 wks, regardless of whether or not the concrete was sealed prior to application or the presence of flour contamination. However, with the low label rate of 10 mg AI/m2, the flour apparently absorbed the insecticide residues from the treated surface, and sealing the concrete did not have a beneficial effect on efficacy. Two field studies with the low label rate were conducted during autumn of 2012 and 2013 and summer of 2013, using only unsealed concrete. Accumulated milling debris caused a reduction in efficacy in the autumn studies, as shown by increased time to 100% knockdown, decreased mortality, and decreased residual efficacy. There was no such corresponding decrease in residual efficacy in the summer study. Overall, results of both studies show that accumulated food and milling debris can absorb residue of beta-cyfluthrin from a treated surface and have a negative impact on residual efficacy, particularly with the low label rate of 10 mg AI/m2. PMID:26470194

  6. Innexin7a forms junctions that stabilize the basal membrane during cellularization of the blastoderm in Tribolium castaneum.

    PubMed

    van der Zee, Maurijn; Benton, Matthew A; Vazquez-Faci, Tania; Lamers, Gerda E M; Jacobs, Chris G C; Rabouille, Catherine

    2015-06-15

    In insects, the fertilized egg undergoes a series of rapid nuclear divisions before the syncytial blastoderm starts to cellularize. Cellularization has been extensively studied in Drosophila melanogaster, but its thick columnar blastoderm is unusual among insects. We therefore set out to describe cellularization in the beetle Tribolium castaneum, the embryos of which exhibit a thin blastoderm of cuboidal cells, like most insects. Using immunohistochemistry, live imaging and transmission electron microscopy, we describe several striking differences to cellularization in Drosophila, including the formation of junctions between the forming basal membrane and the yolk plasmalemma. To identify the nature of this novel junction, we used the parental RNAi technique for a small-scale screen of junction proteins. We find that maternal knockdown of Tribolium innexin7a (Tc-inx7a), an ortholog of the Drosophila gap junction gene Innexin 7, leads to failure of cellularization. In Inx7a-depleted eggs, the invaginated plasma membrane retracts when basal cell closure normally begins. Furthermore, transiently expressed tagged Inx7a localizes to the nascent basal membrane of the forming cells in wild-type eggs. We propose that Inx7a forms the newly identified junctions that stabilize the forming basal membrane and enable basal cell closure. We put forward Tribolium as a model for studying a more ancestral mode of cellularization in insects.

  7. The ABCs of eye color in Tribolium castaneum: orthologs of the Drosophila white, scarlet, and brown Genes.

    PubMed

    Grubbs, Nathaniel; Haas, Sue; Beeman, Richard W; Lorenzen, Marcé D

    2015-03-01

    In Drosophila melanogaster, each of the three paralogous ABC transporters, White, Scarlet and Brown, is required for normal pigmentation of the compound eye. We have cloned the three orthologous genes from the beetle Tribolium castaneum. Conceptual translations of Tribolium white (Tcw), scarlet (Tcst), and brown (Tcbw) are 51, 48, and 32% identical to their respective Drosophila counterparts. We have identified loss-of-eye-pigment strains that bear mutations in Tcw and Tcst: the Tcw gene in the ivory (i) strain carries a single-base transversion, which leads to an E → D amino-acid substitution in the highly conserved Walker B motif, while the Tcst gene in the pearl (p) strain has a deletion resulting in incorporation of a premature stop codon. In light of these findings, the mutant strains i and p are herein renamed white(ivory) (w(i)) and scarlet(pearl) (st(p)), respectively. In addition, RNA inhibition of Tcw and Tcst recapitulates the mutant phenotypes, confirming the roles of these genes in normal eye pigmentation, while RNA interference of Tcbw provides further evidence that it has no role in eye pigmentation in Tribolium. We also consider the evolutionary implications of our findings.

  8. Experimental removal of sexual selection leads to decreased investment in an immune component in female Tribolium castaneum.

    PubMed

    Hangartner, Sandra; Michalczyk, Łukasz; Gage, Matthew J G; Martin, Oliver Y

    2015-07-01

    Because of divergent selection acting on males and females arising from different life-history strategies, polyandry can be expected to promote sexual dimorphism of investment into immune function. In previous work we have established the existence of such divergence within populations where males and females are exposed to varying degrees of polyandry. We here test whether the removal of sexual selection via enforced monogamy generates males and females that have similar levels of investment in immune function. To test this prediction experimentally, we measured differences between the sexes in a key immune measurement (phenoloxidase (PO) activity) and resistance to the microsporidian Paranosema whitei in Tribolium castaneum lines that evolved under monogamous (sexual selection absent) vs polyandrous (sexual selection present) mating systems. At generation 49, all selected lines were simultaneously assessed for PO activity and resistance to their natural parasite P. whitei after two generations of relaxed selection. We found that the polyandrous regime was associated with a clear dimorphism in immune function: females had significantly higher PO activities than males in these lines. In contrast, there was no such difference between the sexes in the lines evolving under the monogamous regime. Survival in the infection experiment did not differ between mating systems or sexes. Removing sexual selection via enforced monogamy thus seems to erase intersexual differences in immunity investment. We suggest that higher PO activities in females that have evolved under sexual selection might be driven by the increased risk of infections and/or injuries associated with exposure to multiple males.

  9. Effects of Flour and Milling Debris on Efficacy of Beta-Cyfluthrin for Control of Tribolium castaneum (Coleoptera: Tenebrionidae).

    PubMed

    Arthur, Frank H; Starkus, Laura A; Mckay, Tanja

    2015-04-01

    Laboratory and field studies were conducted to determine if accumulation of a flour food source or milling debris affected residual efficacy of beta-cyfluthrin for control of Tribolium castaneum (Herbst), the red flour beetle. In the laboratory study, the high label rate of 20 mg active ingredient (AI)/m2 gave effective control for 8 wks, regardless of whether or not the concrete was sealed prior to application or the presence of flour contamination. However, with the low label rate of 10 mg AI/m2, the flour apparently absorbed the insecticide residues from the treated surface, and sealing the concrete did not have a beneficial effect on efficacy. Two field studies with the low label rate were conducted during autumn of 2012 and 2013 and summer of 2013, using only unsealed concrete. Accumulated milling debris caused a reduction in efficacy in the autumn studies, as shown by increased time to 100% knockdown, decreased mortality, and decreased residual efficacy. There was no such corresponding decrease in residual efficacy in the summer study. Overall, results of both studies show that accumulated food and milling debris can absorb residue of beta-cyfluthrin from a treated surface and have a negative impact on residual efficacy, particularly with the low label rate of 10 mg AI/m2.

  10. A Tribolium castaneum whole-embryo culture protocol for studying the molecular mechanisms and morphogenetic movements involved in insect development.

    PubMed

    Macaya, Constanza C; Saavedra, Patricio E; Cepeda, Rodrigo E; Nuñez, Viviana A; Sarrazin, Andres F

    2016-01-01

    The development of the red flour beetle Tribolium castaneum is more representative of arthropods than the evolutionarily derived fly, Drosophila melanogaster. Thus, Tribolium is becoming an emerging organism model for studying the evolution of the mechanisms that control embryonic development in arthropods. In this regard, diverse genetic and molecular tools are currently available for Tribolium, as well as imaging and embryonic techniques. Recently, we developed a method for culturing embryos in order to study specific stages during Tribolium development. In this report, we present a detailed and "easy-to-follow" protocol for embryo handling and dissection, extending the use of whole-embryo culture to functional analysis by performing in vivo pharmacological manipulations. This experimental accessibility allowed us to study the relevance of microtubules in axis elongation, using nocodazole and taxol drugs to interfere with microtubule networks, followed by length measurement analysis. Additionally, we demonstrated that embryo handling had no effect on the development of Tribolium embryos, and we checked viability after dissection and bisection and during incubation using propidium iodide. The embryo culture protocol we describe here can be applied to study diverse developmental processes in Tribolium. We expect that this protocol can be adapted and applied to other arthropods.

  11. Repellent and Fumigant Activities of Tanacetum nubigenum Wallich. ex DC Essential Oils against Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae).

    PubMed

    Haider, S Zafar; Mohan, Manindra; Pandey, Abhay K; Singh, Pooja

    2015-01-01

    The repellent and fumigant toxicity of essential oils of Tanacetum nubigenum Wallich. ex DC collected from three different habitats (Gothing, Burphu and Glacier) of Uttarakhand Himalayas, India named as TNG, TNB and TNM respectively, were investigated against the adults of red flour beetle Tribolium castaneum (Herbst). Among the three samples tested, TNG was found to more potent exhibiting more repellent effect towards the insects and with LC50 values by fumigant bioassay were 13.23 and 8.32 µl per 0.25 L air at 24 and 48 h exposure of insects to the essential oil respectively. In between other two oil samples, TNM was superior in potency showed LC50 value of 14.22 (24 h) & 8.82 µl per 0.25 L air (48 h). During in vivo study all the essential oil samples significantly protected 500 g of wheat grains for 6 months from insect infestation as compared to non fumigated grains and order of efficacy was TNG>TNM>TNB. There were no side effects of essential oils on germination rate of grains (<85%) exposed for 6 months after fumigation. The present study suggests that essential oil of T. nubigenum can be explored as novel natural fumigants for the control of stored product insects. PMID:26235002

  12. Cholinergic and non-cholinergic functions of two acetylcholinesterase genes revealed by gene-silencing in Tribolium castaneum

    PubMed Central

    Lu, Yanhui; Park, Yoonseong; Gao, Xiwu; Zhang, Xin; Yao, Jianxiu; Pang, Yuan-Ping; Jiang, Haobo; Zhu, Kun Yan

    2012-01-01

    We compared biological functions of two acetylcholinesterase genes (TcAce1 and TcAce2) in Tribolium castaneum, a globally distributed major pest of stored grain products and an emerging model organism, by using RNA interference. Although both genes expressed at all developmental stages and mainly in the brain, the transcript level of TcAce1 was 1.2- to 8.7-fold higher than that of TcAce2, depending on developmental stages. Silencing TcAce1 in 20-day larvae led to 100% mortality within two weeks after eclosion and increased larval susceptibilities to anticholinesterase insecticides. In contrast, silencing TcAce2 did not show insect mortality and significantly affect insecticide susceptibility, but delayed insect development and reduced female egg-laying and egg hatching. These results demonstrate for the first time that TcAce1 plays a major role in cholinergic functions and is the target of anticholinesterase insecticides, whereas TcAce2 plays an important, non-cholinergic role in female reproduction, embryo development, and growth of offspring. PMID:22371826

  13. Differences in Attack Avoidance and Mating Success between Strains Artificially Selected for Dispersal Distance in Tribolium castaneum

    PubMed Central

    Matsumura, Kentarou; Miyatake, Takahisa

    2015-01-01

    Individuals of both dispersal and non-dispersal types (disperser and non-disperser) are found in a population, suggesting that each type has both costs and benefits for fitness. However, few studies have examined the trade-off between the costs and benefits for the types. Here, we artificially selected for walking distance, i.e., an indicator of dispersal ability, in the red flour beetle Tribolium castaneum and established strains with longer (L-strains) or shorter (S-strains) walking distances. We then compared the frequency of predation by the assassin bug Amphibolus venator and the mating frequency of the selected strains. L-strain beetles suffered higher predation risk, than did S-strain beetles. L-strain males had significantly increased mating success compared to S-strain males, but females did not show a significant difference between the strains. The current results showed the existence of a trade-off between predation avoidance and mating success associated with dispersal types at a genetic level only in males. This finding can help to explain the maintenance of variation in dispersal ability within a population. PMID:25970585

  14. The role of epigenetics in host-parasite coevolution: lessons from the model host insects Galleria mellonella and Tribolium castaneum.

    PubMed

    Vilcinskas, Andreas

    2016-08-01

    Recent studies addressing experimental host-parasite coevolution and transgenerational immune priming in insects provide evidence for heritable shifts in host resistance or parasite virulence. These rapid reciprocal adaptations may thus be transferred to offspring generations by either genetic changes or mechanisms that do not involve changes in the germline DNA sequence. Epigenetic inheritance refers to changes in gene expression that are heritable across generations and mediated by epigenetic modifications passed from parents to offspring. Highlighting the role of epigenetics in host-parasite coevolution, this review discusses the involvement of DNA methylation, histone acetylation/deacetylation and microRNAs in the interactions between bacterial or fungal parasites and model host insects such as the greater wax moth Galleria mellonella and the red flour beetle Tribolium castaneum. These epigenetic mechanisms are thought to participate in generation-spanning transcriptional reprogramming in the host insect, often linking immunity with developmentally related gene expression and contributing to the heredity of acquired adaptations. It is proposed that the interactions during host-parasite coevolution can therefore be expanded beyond reciprocal genetic changes to include reciprocal epigenetic changes. Epigenetics is thus a promising and prospering field in the context of host-parasite coevolution.

  15. Sodium Solute Symporter and Cadherin Proteins Act as Bacillus thuringiensis Cry3Ba Toxin Functional Receptors in Tribolium castaneum*

    PubMed Central

    Contreras, Estefanía; Schoppmeier, Michael; Real, M. Dolores; Rausell, Carolina

    2013-01-01

    Understanding how Bacillus thuringiensis (Bt) toxins interact with proteins in the midgut of susceptible coleopteran insects is crucial to fully explain the molecular bases of Bt specificity and insecticidal action. In this work, aminopeptidase N (TcAPN-I), E-cadherin (TcCad1), and sodium solute symporter (TcSSS) have been identified by ligand blot as putative Cry3Ba toxin-binding proteins in Tribolium castaneum (Tc) larvae. RNA interference knockdown of TcCad1 or TcSSS proteins resulted in decreased susceptibility to Cry3Ba toxin, demonstrating the Cry toxin receptor functionality for these proteins. In contrast, TcAPN-I silencing had no effect on Cry3Ba larval toxicity, suggesting that this protein is not relevant in the Cry3Ba toxin mode of action in Tc. Remarkable features of TcSSS protein were the presence of cadherin repeats in its amino acid sequence and that a TcSSS peptide fragment containing a sequence homologous to a binding epitope found in Manduca sexta and Tenebrio molitor Bt cadherin functional receptors enhanced Cry3Ba toxicity. This is the first time that the involvement of a sodium solute symporter protein as a Bt functional receptor has been demonstrated. The role of this novel receptor in Bt toxicity against coleopteran insects together with the lack of receptor functionality of aminopeptidase N proteins might account for some of the differences in toxin specificity between Lepidoptera and Coleoptera insect orders. PMID:23645668

  16. The ABCs of eye color in Tribolium castaneum: orthologs of the Drosophila white, scarlet, and brown Genes.

    PubMed

    Grubbs, Nathaniel; Haas, Sue; Beeman, Richard W; Lorenzen, Marcé D

    2015-03-01

    In Drosophila melanogaster, each of the three paralogous ABC transporters, White, Scarlet and Brown, is required for normal pigmentation of the compound eye. We have cloned the three orthologous genes from the beetle Tribolium castaneum. Conceptual translations of Tribolium white (Tcw), scarlet (Tcst), and brown (Tcbw) are 51, 48, and 32% identical to their respective Drosophila counterparts. We have identified loss-of-eye-pigment strains that bear mutations in Tcw and Tcst: the Tcw gene in the ivory (i) strain carries a single-base transversion, which leads to an E → D amino-acid substitution in the highly conserved Walker B motif, while the Tcst gene in the pearl (p) strain has a deletion resulting in incorporation of a premature stop codon. In light of these findings, the mutant strains i and p are herein renamed white(ivory) (w(i)) and scarlet(pearl) (st(p)), respectively. In addition, RNA inhibition of Tcw and Tcst recapitulates the mutant phenotypes, confirming the roles of these genes in normal eye pigmentation, while RNA interference of Tcbw provides further evidence that it has no role in eye pigmentation in Tribolium. We also consider the evolutionary implications of our findings. PMID:25555987

  17. The roles of thermal transient receptor potential channels in thermotactic behavior and in thermal acclimation in the red flour beetle, Tribolium castaneum.

    PubMed

    Kim, Hong Geun; Margolies, David; Park, Yoonseong

    2015-05-01

    To survive in variable or fluctuating temperature, organisms should show appropriate behavioral and physiological responses which must be mediated through properly attuned thermal sensory mechanisms. Transient receptor potential channels (TRPs) are a family of cation channels a number of which, called thermo-TRPs, are known to function as thermosensors. We investigated the potential role of thermo-TPRs that have been previously identified in the fruit fly, Drosophila melanogaster, in thermotaxis and thermal acclimation in the red flour beetle, Tribolium castaneum. Phylogenetic analysis of the trp genes showed generally one-to-one orthology between those in D. melanogaster and in T. castaneum, although there are putative gene-losses in two TRP subfamilies of D. melanogaster. With RNA interference (RNAi) of T. castaneum thermo-TRP candidates painless, pyrexia and trpA1, we measured thermal avoidance behavior. RNAi of trpA1 resulted in reduced avoidance of high temperatures, 39 and 42 °C. We also measured the effects of RNAi on heat-induced knockout and death under a short exposure to high temperature (1min at 52 °C) either with or without a 10-min acclimation period at 42 °C. Relatively short exposure to high temperature was enough to induce high temperature thermal acclimation. RNAi of trpA1 led to faster knockout at 52 °C. RNAi of painless showed lower recovery rates from heat-induced knockout after thermal acclimation, and RNAi of pyrexia showed lower long-term survivorship without thermal acclimation. Therefore, we concluded that trpA1 is important in high temperature sensing and also in enhanced tolerance to high-temperature induced knockout; painless plays a role in rapid acclimation to high temperature; and pyrexia functions in protecting beetles from acute heat stress without acclimation.

  18. Dissecting systemic RNA interference in the red flour beetle Tribolium castaneum: parameters affecting the efficiency of RNAi.

    PubMed

    Miller, Sherry C; Miyata, Keita; Brown, Susan J; Tomoyasu, Yoshinori

    2012-01-01

    The phenomenon of RNAi, in which the introduction of dsRNA into a cell triggers the destruction of the corresponding mRNA resulting in a gene silencing effect, is conserved across a wide array of plant and animal phyla. However, the mechanism by which the dsRNA enters a cell, allowing the RNAi effect to occur throughout a multicellular organism (systemic RNAi), has only been studied extensively in certain plants and the nematode Caenorhabditis elegans. In recent years, RNAi has become a popular reverse genetic technique for gene silencing in many organisms. Although many RNAi techniques in non-traditional model organisms rely on the systemic nature of RNAi, little has been done to analyze the parameters required to obtain a robust systemic RNAi response. The data provided here show that the concentration and length of dsRNA have profound effects on the efficacy of the RNAi response both in regard to initial efficiency and duration of the effect in Tribolium castaneum. In addition, our analyses using a series of short dsRNAs and chimeric dsRNA provide evidence that dsRNA cellular uptake (and not the RNAi response itself) is the major step affected by dsRNA size in Tribolium. We also demonstrate that competitive inhibition of dsRNA can occur when multiple dsRNAs are injected together, influencing the effectiveness of RNAi. These data provide specific information essential to the design and implementation of RNAi based studies, and may provide insight into the molecular basis of the systemic RNAi response in insects. PMID:23133513

  19. Contact Toxicity of Deltamethrin Against Tribolium castaneum (Coleoptera: Tenebrionidae), Sitophilus oryzae (Coleoptera: Curculionidae), and Rhyzopertha dominica (Coleoptera: Bostrichidae) Adults.

    PubMed

    Paudyal, S; Opit, G P; Arthur, F H; Bingham, G V; Gautam, S G

    2016-08-01

    This study was conducted at Oklahoma State University, Stillwater, OK, to evaluate the response to deltamethrin concentrations for adults of three stored-product insects, Tribolium castaneum (Herbst), Sitophilus oryzae (L.), and Rhyzopertha dominica (F.). In insect toxicological studies, knockdown is the state of intoxication and partial paralysis as a result of exposure to an insecticide. Deltamethrin concentrations ranging from 0.48 to 140 mg/m(2) (1 to 3,000 ppm) were sprayed on glass Petri dishes. After the dishes dried, 20 adult insects of each species were placed on the treated dishes to determine the contact toxicity of deltamethrin. Assessments for knockdown were made at 15-min intervals for up to 8 h after initial exposure and then again after 24 or 48 h. Insects were then transferred to clean untreated Petri dishes with diet and observed from 0.5 to 72 h. Mortality was assessed 72 h after transfer to untreated dishes with food material. Deltamethrin was highly effective against all three species tested and achieved 99% knockdown of insects of all species within 4 h after exposure at concentrations ≥1.2 mg/m(2) Although some insects recovered from initial knockdown at concentrations ≤48 mg/m(2), nearly all the insects were killed at 140 mg/m(2) when exposed for 48 h. LC95 values for all species tested, for the 48-h exposure period, were <140 mg/m(2), the concentration of deltamethrin that could potentially be present in new ZeroFly Storage Bag fabric. ZeroFly bags are used for stored-product insect pest control.

  20. A temperature shock can lead to trans-generational immune priming in the Red Flour Beetle, Tribolium castaneum

    PubMed Central

    Eggert, Hendrik; Diddens-de Buhr, Maike F; Kurtz, Joachim

    2015-01-01

    Trans-generational immune priming (TGIP) describes the transfer of immune stimulation to the next generation. As stress and immunity are closely connected, we here address the question whether trans-generational effects on immunity and resistance can also be elicited by a nonpathogen stress treatment of parents. General stressors have been shown to induce immunity to pathogens within individuals. However, to our knowledge, it is as of yet unknown whether stress can also induce trans-generational effects on immunity and resistance. We exposed a parental generation (mothers, fathers, or both parents) of the red flour beetle Tribolium castaneum, a species where TGIP has been previously been demonstrated, to either a brief heat or cold shock and examined offspring survival after bacterial infection with the entomopathogen Bacillus thuringiensis. We also studied phenoloxidase activity, a key enzyme of the insect innate immune system that has previously been demonstrated to be up-regulated upon TGIP. We quantified parental fecundity and offspring developmental time to evaluate whether trans-generational priming might have costs. Offspring resistance was found to be significantly increased when both parents received a cold shock. Offspring phenoloxidase activity was also higher when mothers or both parents were cold-shocked. By contrast, parental heat shock reduced offspring phenoloxidase activity. Moreover, parental cold or heat shock delayed offspring development. In sum, we conclude that trans-generational priming for resistance could not only be elicited by pathogens or pathogen-derived components, but also by more general cues that are indicative of a stressful environment. The interaction between stress responses and the immune system might play an important role also for trans-generational effects. PMID:25859336

  1. Contact Toxicity of Deltamethrin Against Tribolium castaneum (Coleoptera: Tenebrionidae), Sitophilus oryzae (Coleoptera: Curculionidae), and Rhyzopertha dominica (Coleoptera: Bostrichidae) Adults.

    PubMed

    Paudyal, S; Opit, G P; Arthur, F H; Bingham, G V; Gautam, S G

    2016-08-01

    This study was conducted at Oklahoma State University, Stillwater, OK, to evaluate the response to deltamethrin concentrations for adults of three stored-product insects, Tribolium castaneum (Herbst), Sitophilus oryzae (L.), and Rhyzopertha dominica (F.). In insect toxicological studies, knockdown is the state of intoxication and partial paralysis as a result of exposure to an insecticide. Deltamethrin concentrations ranging from 0.48 to 140 mg/m(2) (1 to 3,000 ppm) were sprayed on glass Petri dishes. After the dishes dried, 20 adult insects of each species were placed on the treated dishes to determine the contact toxicity of deltamethrin. Assessments for knockdown were made at 15-min intervals for up to 8 h after initial exposure and then again after 24 or 48 h. Insects were then transferred to clean untreated Petri dishes with diet and observed from 0.5 to 72 h. Mortality was assessed 72 h after transfer to untreated dishes with food material. Deltamethrin was highly effective against all three species tested and achieved 99% knockdown of insects of all species within 4 h after exposure at concentrations ≥1.2 mg/m(2) Although some insects recovered from initial knockdown at concentrations ≤48 mg/m(2), nearly all the insects were killed at 140 mg/m(2) when exposed for 48 h. LC95 values for all species tested, for the 48-h exposure period, were <140 mg/m(2), the concentration of deltamethrin that could potentially be present in new ZeroFly Storage Bag fabric. ZeroFly bags are used for stored-product insect pest control. PMID:27270576

  2. Insecticide-Mediated Up-Regulation of Cytochrome P450 Genes in the Red Flour Beetle (Tribolium castaneum)

    PubMed Central

    Liang, Xiao; Xiao, Da; He, Yanping; Yao, Jianxiu; Zhu, Guonian; Zhu, Kun Yan

    2015-01-01

    Some cytochrome P450 (CYP) genes are known for their rapid up-regulation in response to insecticide exposures in insects. To date, however, limited information is available with respect to the relationships among the insecticide type, insecticide concentration, exposure duration and the up-regulated CYP genes. In this study, we examined the transcriptional response of eight selected CYP genes, including CYP4G7, CYP4Q4, CYP4BR3, CYP12H1, CYP6BK11, CYP9D4, CYP9Z5 and CYP345A1, to each of four insecticides in the red flour beetle, Tribolium castaneum. Reverse transcription quantitative PCR (RT-qPCR) revealed that CYP4G7 and CYP345A1 can be significantly up-regulated by cypermethrin (1.97- and 2.06-fold, respectively), permethrin (2.00- and 2.03-fold) and lambda-cyhalothrin (1.73- and 1.81-fold), whereas CYP4BR3 and CYP345A1 can be significantly up-regulated by imidacloprid (1.99- and 1.83-fold) when 20-day larvae were exposed to each of these insecticides at the concentration of LC20 for 24 h. Our studies also showed that similar levels of up-regulation can be achieved for CYP4G7, CYP4BR3 and CYP345A1 by cypermethrin, permethrin, lambda-cyhalothrin or imidacloprid with approximately one fourth of LC20 in 6 h. Our study demonstrated that up-regulation of these CYP genes was rapid and only required low concentrations of insecticides, and the up-regulation not only depended on the CYP genes but also the type of insecticides. Our results along with those from previous studies also indicated that there were no specific patterns for predicting the up-regulation of specific CYP gene families based on the insecticide classification. PMID:25607733

  3. Insecticide-mediated up-regulation of cytochrome P450 genes in the red flour beetle (Tribolium castaneum).

    PubMed

    Liang, Xiao; Xiao, Da; He, Yanping; Yao, Jianxiu; Zhu, Guonian; Zhu, Kun Yan

    2015-01-19

    Some cytochrome P450 (CYP) genes are known for their rapid up-regulation in response to insecticide exposures in insects. To date, however, limited information is available with respect to the relationships among the insecticide type, insecticide concentration, exposure duration and the up-regulated CYP genes. In this study, we examined the transcriptional response of eight selected CYP genes, including CYP4G7, CYP4Q4, CYP4BR3, CYP12H1, CYP6BK11, CYP9D4, CYP9Z5 and CYP345A1, to each of four insecticides in the red flour beetle, Tribolium castaneum. Reverse transcription quantitative PCR (RT-qPCR) revealed that CYP4G7 and CYP345A1 can be significantly up-regulated by cypermethrin (1.97- and 2.06-fold, respectively), permethrin (2.00- and 2.03-fold) and lambda-cyhalothrin (1.73- and 1.81-fold), whereas CYP4BR3 and CYP345A1 can be significantly up-regulated by imidacloprid (1.99- and 1.83-fold) when 20-day larvae were exposed to each of these insecticides at the concentration of LC20 for 24 h. Our studies also showed that similar levels of up-regulation can be achieved for CYP4G7, CYP4BR3 and CYP345A1 by cypermethrin, permethrin, lambda-cyhalothrin or imidacloprid with approximately one fourth of LC20 in 6 h. Our study demonstrated that up-regulation of these CYP genes was rapid and only required low concentrations of insecticides, and the up-regulation not only depended on the CYP genes but also the type of insecticides. Our results along with those from previous studies also indicated that there were no specific patterns for predicting the up-regulation of specific CYP gene families based on the insecticide classification.

  4. The role of Broad in the development of Tribolium castaneum: implications for the evolution of the holometabolous insect pupa.

    PubMed

    Suzuki, Yuichiro; Truman, James W; Riddiford, Lynn M

    2008-02-01

    The evolution of complete metamorphosis in insects is a key innovation that has led to the successful diversification of holometabolous insects, yet the origin of the pupa remains an enigma. Here, we analyzed the expression of the pupal specifier gene broad (br), and the effect on br of isoform-specific, double-stranded RNA-mediated silencing, in a basal holometabolous insect, the beetle Tribolium castaneum. All five isoforms are weakly expressed during the penultimate instar and highly expressed during the prepupal period of the final instar. Application of hydroprene, a juvenile hormone analog, during the penultimate instar caused a repeat of the penultimate br expression patterns, and the formation of supernumerary larvae. Use of dsRNA against the br core region, or against a pair of either the br-Z2 or br-Z3 isoform with the br-Z1 or br-Z4 isoform, produced mobile animals with well-differentiated adult-like appendages, but which retained larval-like urogomphi and epidermis. Disruption of either the br-Z2 or the br-Z3 isoform caused the formation of shorter wings. Disruption of both br-Z1 and br-Z4 caused the appearance of pupal traits in the adults, but disruption of br-Z5 had no morphological effect. Our findings show that the br isoform functions are broadly conserved within the Holometabola and suggest that evolution of br isoform expression may have played an important role in the evolution of the pupa in holometabolous insects.

  5. Expressional and functional analysis of CYP15A1, a juvenile hormone epoxidase, in the red flour beetle Tribolium castaneum.

    PubMed

    Minakuchi, Chieka; Ishii, Fumika; Washidu, Yumiko; Ichikawa, Akio; Tanaka, Toshiharu; Miura, Ken; Shinoda, Tetsuro

    2015-09-01

    Juvenile hormone (JH) is synthesized and secreted by the corpora allata. In the final two steps of JH biosynthesis, farnesoic acid (FA) is converted to JH through methylation by JH acid O-methyltransferase (JHAMT) and epoxidation by the cytochrome P450 enzyme CYP15. In the present study, we identified a homolog of CYP15 from the red flour beetle Tribolium castaneum (TcCYP15A1), and analyzed its expression as well as its role in JH biosynthesis. Quantitative RT-PCR analysis showed that the level of TcCYP15A1 mRNA was high in the embryonic stage as well as in the middle of the final larval instar. In the embryonic stage, the transcript level of TcCYP15A1 started to increase 30h after egg laying (AEL), peaked 54-60h AEL, and was followed by an increase of TcJHAMT mRNA, suggesting that JH biosynthesis started at this time point. TcCYP15A1 mRNA was present, but not exclusively so in the larval corpora allata. The recombinant TcCYP15A1 protein epoxidized both FA and methyl farnesoate (MF) in highly stereo-specific manners. These results confirmed that TcCYP15A1 is involved in JH biosynthesis. The RNAi-mediated knockdown of TcCYP15A1 in the pre-final larval instar did not result in precocious metamorphosis to pupa, indicating that MF may exhibit JH-like activity in order to maintain the larval status. The double knockdown of TcJHAMT and TcCYP15A1 resulted in pupae and adults with shorter wings, suggesting that the precursors of JH, JH acid and MF, may be essential for wing expansion.

  6. A Major Facilitator Superfamily protein encoded by TcMucK gene is not required for cuticle pigmentation, growth and development in Tribolium castaneum.

    PubMed

    Mun, Seulgi; Noh, Mi Young; Osanai-Futahashi, Mizuko; Muthukrishnan, Subbaratnam; Kramer, Karl J; Arakane, Yasuyuki

    2014-06-01

    Insect cuticle pigmentation and sclerotization (tanning) are vital physiological processes for insect growth, development and survival. We have previously identified several colorless precursor molecules as well as enzymes involved in their biosynthesis and processing to yield the mature intensely colored body cuticle pigments. A recent study indicated that the Bombyx mori (silkmoth) gene, BmMucK, which encodes a protein orthologous to a Culex pipiens quiquefasciatus (Southern house mosquito) cis,cis, muconate transporter, is a member of the "Major Facilitator Superfamily" (MFS) of transporter proteins and is associated with the appearance of pigmented body segments of naturally occurring body color mutants of B. mori. While RNA interference of the BmMucK gene failed to result in any observable phenotype, RNAi using a dsRNA for an orthologous gene from the red flour beetle, Tribolium castaneum, was reported to result in molting defects and darkening of the cuticle and some body parts, leading to the suggestion that orthologs of MucK genes may differ in their functions among insects. To verify the role and essentiality of the ortholog of this gene in development and body pigmentation function in T. castaneum we obtained cDNAs for the orthologous gene (TcMucK) from RNA isolated from the GA-1 wild-type strain of T. castaneum. The sequence of a 1524 nucleotides-long cDNA for TcMucK which encodes the putatively full-length protein, was assembled from two overlapping RT-PCR fragments and the expression profile of this gene during development was analyzed by real-time PCR. This cDNA encodes a 55.8 kDa protein consisting of 507 amino acid residues and includes 11 putative transmembrane segments. Transcripts of TcMucK were detected throughout all of the developmental stages analyzed. The function of this gene was explored by injection of two different double-stranded RNAs targeting different regions of the TcMucK gene (dsTcMucKs) into young larvae to down

  7. Morpho-functional characterization and esterase patterns of the midgut of Tribolium castaneum Herbst, 1797 (Coleoptera: Tenebrionidae) parasitized by Gregarina cuneata (Apicomplexa: Eugregarinidae).

    PubMed

    Gigliolli, Adriana A Sinópolis; Lapenta, Ana Silva; Ruvolo-Takasusuki, Maria Claudia Colla; Abrahão, Josielle; Conte, Hélio

    2015-09-01

    Tribolium castaneum (Coleoptera: Tenebrionidae) is a common pest of stored grains and byproducts and is normally infected by Gregarina cuneata (Apicomplexa: Eugregarinidae). The life cycle of this parasite includes the sporozoite, trophozoite, gamont, gametocyte, and oocyst stages, which occur between the epithelium and lumen of the host's midgut. This study aims to describe the morphofunctional alterations in the midgut and determine the esterase patterns in T. castaneum when parasitized by gregarines. To achieve this purpose, midguts of adult insects were isolated, processed, and analysed using light and electron microscopy. We determined total protein content, amylase activity, and the expression and related activities of the esterases by using polyacrylamide gel electrophoresis (PAGE). The midgut of T. castaneum is formed by digestive, regenerative, and endocrine cells. The effects of parasitism on the digestive cells are severe, because the gregarines remain attached to these cells to absorb all the nutrients they need throughout their development. In these cells, the most common alterations observed include expansion and fragmentation of the rough endoplasmic reticulum, development of the smooth endoplasmic reticulum, changes in mitochondrial cristae, cytoplasmic vacuolization, formation of myelin structures, spherites, large intercellular spaces, autophagic vesicles, expansion of the basal labyrinth, and cytoplasmic protrusions. Deposits of glycogen granules were also observed. Amylase activity was reduced in parasitized insects. Regenerative cells were found in disorganized crypts and did not differentiate into new cells, thus, compromising the restoration of the damaged epithelium. Though few morphological alterations were observed in the endocrine cells, results suggest that the synthesis and/or release of hormones might be impaired. Nine esterases (EST-1 to 9) were identified in the midgut of T. castaneum and were expressed in varying levels in response

  8. Tribolium castaneum immune defense genes are differentially expressed in response to Bacillus thuringiensis toxins sharing common receptor molecules and exhibiting disparate toxicity.

    PubMed

    Contreras, Estefanía; Benito-Jardón, María; López-Galiano, M José; Real, M Dolores; Rausell, Carolina

    2015-06-01

    In Tribolium castaneum larvae we have demonstrated by RNA interference knockdown that the Bacillus thuringiensis Cry3Ba toxin receptors Cadherin-like and Sodium solute symporter proteins are also functional receptors of the less active Cry3Aa toxin. Differences in susceptibility to B. thuringiensis infection might not only rely on toxin-receptor interaction but also on host defense mechanisms. We compared the expression of the immune related genes encoding Apolipophorin-III and two antimicrobial peptides, Defensin3 and Defensin2 after B. thuringiensis challenge. All three genes were up-regulated following Cry3Ba spore-crystal intoxication whereas only Defensins gene expression was induced upon Cry3Aa spore-crystal treatment, evidencing a possible association between host immune response and larval susceptibility to B. thuringiensis. We assessed the antimicrobial activity spectra of T. castaneum defensins peptide fragments and found that a peptide fragment of Defensin3 was effective against the human microbial pathogens, Escherichia coli, Staphylococcus aureus and Candida albicans, being S. aureus the most susceptible one.

  9. The life cycle of Gregarina cuneata in the midgut of Tribolium castaneum and the effects of parasitism on the development of insects.

    PubMed

    Gigliolli, A A S; Julio, A H F; Conte, H

    2016-04-01

    Tribolium castaneum Herbst 1797 (Coleoptera: Tenebrionidae), an important pest of stored grains and byproducts, is naturally infected by Gregarina cuneata Stein 1848 (Apicomplexa: Gregarinidae). Changes in the life cycle of insects caused by the parasite development in the midgut were studied. Trophozoites, gamonts (solitary and associated), and gametocysts were present in the midgut of the insects. In young trophozoites, the apical region differentiated into an epimerite that firmly attached the parasite to the host epithelial cells. With maturation, trophozoites developed in gamonts that were associated with the initiation of sexual reproduction in the cell cycle, culminating in the formation of the spherical gametocyst. Morpho-functional analyses indicated that gregarines absorb nutrients from infected cells and can occlude the midgut as they develop. Consequently, nutritional depletion may interfere with the host's physiology, causing decreased growth, delayed development, and high mortality rates of the parasitized insects. These results suggest G. cuneata could be an important biological agent for controlling T. castaneum in integrated pest management programs.

  10. Transcription factor broad suppresses precocious development of adult structures during larval-pupal metamorphosis in the red flour beetle, Tribolium castaneum.

    PubMed

    Parthasarathy, R; Tan, A; Bai, H; Palli, Subba R

    2008-01-01

    Broad (br), a transcription factor containing the Broad-Tramtrack-Bric-a-brac (BTB) and zinc finger domains was shown to mediate 20-hydroxyecdysone (20E) action and pupal development in Drosophila melanogaster and Manduca sexta. We determined the key roles of br during larval-pupal metamorphosis using RNA interference (RNAi) in a coleopteran insect, Tribolium castaneum. Two major peaks of T. castaneum broad (Tcbr) mRNA, one peak at the end of feeding stage prior to the larvae entering the quiescent stage and another peak during the quiescent stage were detected in the whole body and midgut tissue dissected from staged insects. Expression of br during the final instar larval stage is essential for successful larval-pupal metamorphosis, because, RNAi-mediated knock-down of Tcbr during this stage derailed larval-pupal metamorphosis and produced insects that showed larval, pupal and adult structures. Tcbr dsRNA injected into the final instar larvae caused reduction in the mRNA levels of genes known to be involved in 20E action (EcRA, E74 and E75B). Tcbr dsRNA injected into the final instar larvae also caused an increase in the mRNA levels of JH-response genes (JHE and Kr-h1b). Knock-down of Tcbr expression also affected 20E-mediated remodeling of midgut during larval-pupal metamorphosis. These data suggest that the expression of Tcbr during the final instar larval stage promotes pupal program while suppressing the larval and adult programs ensuring a transitory pupal stage in holometabolous insects. PMID:18083350

  11. Hox genes require homothorax and extradenticle for body wall identity specification but not for appendage identity specification during metamorphosis of Tribolium castaneum.

    PubMed

    Smith, Frank W; Jockusch, Elizabeth L

    2014-11-01

    The establishment of segment identity is a key developmental process that allows for divergence along the anteroposterior body axis in arthropods. In Drosophila, the identity of a segment is determined by the complement of Hox genes it expresses. In many contexts, Hox transcription factors require the protein products of extradenticle (exd) and homothorax (hth) as cofactors to perform their identity specification functions. In holometabolous insects, segment identity may be specified twice, during embryogenesis and metamorphosis. To glean insight into the relationship between embryonic and metamorphic segmental identity specification, we have compared these processes in the flour beetle Tribolium castaneum, which develops ventral appendages during embryogenesis that later metamorphose into adult appendages with distinct morphologies. At metamorphosis, comparisons of RNAi phenotypes indicate that Hox genes function jointly with Tc-hth and Tc-exd to specify several region-specific aspects of the adult body wall. On the other hand, Hox genes specify appendage identities along the anteroposterior axis independently of Tc-hth/Tc-exd and Tc-hth/Tc-exd specify proximal vs. distal identity within appendages independently of Hox genes during this stage. During embryogenesis, Tc-hth and Tc-exd play a broad role in the segmentation process and are required for specification of body wall identities in the thorax; however, contrasting with results from other species, we did not obtain homeotic transformations of embryonic appendages in response to Tc-hth or Tc-exd RNAi. In general, the homeotic effects of interference with the function of Hox genes and Tc-hth/Tc-exd during metamorphosis did not match predictions based on embryonic roles of these genes. Comparing metamorphic patterning in T. castaneum to embryonic and post-embryonic development in hemimetabolous insects suggests that holometabolous metamorphosis combines patterning processes of both late embryogenesis and

  12. Two major cuticular proteins are required for assembly of horizontal laminae and vertical pore canals in rigid cuticle of Tribolium castaneum.

    PubMed

    Noh, Mi Young; Kramer, Karl J; Muthukrishnan, Subbaratnam; Kanost, Michael R; Beeman, Richard W; Arakane, Yasuyuki

    2014-10-01

    The insect exoskeleton is composed of cuticle primarily formed from structural cuticular proteins (CPs) and the polysaccharide chitin. Two CPs, TcCPR27 and TcCPR18, are major proteins present in the elytron (highly sclerotized and pigmented modified forewing) as well as the pronotum (dorsal sclerite of the prothorax) and ventral abdominal cuticle of the red flour beetle, Tribolium castaneum. Both CPs belong to the CPR family, which includes proteins that have an amino acid sequence motif known as the Rebers & Riddiford (R&R) consensus sequence. Injection of double-stranded RNA (dsRNA) for TcCPR27 and TcCPR18 resulted in insects with shorter, wrinkled, warped and less rigid elytra than those from control insects. To gain a more comprehensive understanding of the roles of CPs in cuticle assembly, we analyzed for the precise localization of TcCPR27 and the ultrastructural architecture of cuticle in TcCPR27- and TcCPR18-deficient elytra. Transmission electron microscopic analysis combined with immunodetection using gold-labeled secondary antibody revealed that TcCPR27 is present in dorsal elytral procuticle both in the horizontal laminae and in vertical pore canals. dsRNA-mediated RNA interference (RNAi) of TcCPR27 resulted in abnormal electron-lucent laminae and pore canals in elytra except for the boundary between these two structures in which electron-dense molecule(s) apparently accumulated. Insects subjected to RNAi for TcCPR18 also had disorganized laminae and pore canals in the procuticle of elytra. Similar ultrastructural defects were also observed in other body wall regions with rigid cuticle such as the thorax and legs of adult T. castaneum. TcCPR27 and TcCPR18 are required for proper formation of the horizontal chitinous laminae and vertical pore canals that are critical for formation and stabilization of rigid adult cuticle.

  13. A brain-specific cytochrome P450 responsible for the majority of deltamethrin resistance in the QTC279 strain of Tribolium castaneum

    PubMed Central

    Zhu, Fang; Parthasarathy, R.; Bai, Hua; Woithe, Katharina; Kaussmann, Martin; Nauen, Ralf; Harrison, Douglas A.; Palli, Subba R.

    2010-01-01

    Cytochrome P450-mediated detoxification is one of the most important mechanisms involved in insecticide resistance. However, the molecular basis of this mechanism and the physiological functions of P450s associated with insecticide resistance remain largely unknown. Here, we exploited the functional genomics and reverse genetic approaches to identify and characterize a P450 gene responsible for the majority of deltamethrin resistance observed in the QTC279 strain of Tribolium castaneum. We used recently completed whole-genome sequence of T. castaneum to prepare custom microarrays and identified a P450 gene, CYP6BQ9, which showed more than a 200-fold higher expression in the deltamethrin-resistant QTC279 strain when compared with its expression in the deltamethrin-susceptible Lab-S strain. Functional studies using both double-strand RNA (dsRNA)-mediated knockdown in the expression of CYP6BQ9 and transgenic expression of CYP6BQ9 in Drosophila melanogaster showed that CYP6BQ9 confers deltamethrin resistance. Furthermore, CYP6BQ9 enzyme expressed in baculovirus metabolizes deltamethrin to 4-hydroxy deltamethrin. Strikingly, we also found that unlike many P450 genes involved in insecticide resistance that were reported previously, CYP6BQ9 is predominantly expressed in the brain, a part of the central nervous system (CNS) containing voltage-gated sodium channels targeted by deltamethrin. Taken together, the current studies on the brain-specific insect P450 involved in deltamethrin resistance shed new light on the understanding of the molecular basis and evolution of insecticide resistance. PMID:20410462

  14. A brain-specific cytochrome P450 responsible for the majority of deltamethrin resistance in the QTC279 strain of Tribolium castaneum.

    PubMed

    Zhu, Fang; Parthasarathy, R; Bai, Hua; Woithe, Katharina; Kaussmann, Martin; Nauen, Ralf; Harrison, Douglas A; Palli, Subba R

    2010-05-11

    Cytochrome P450-mediated detoxification is one of the most important mechanisms involved in insecticide resistance. However, the molecular basis of this mechanism and the physiological functions of P450s associated with insecticide resistance remain largely unknown. Here, we exploited the functional genomics and reverse genetic approaches to identify and characterize a P450 gene responsible for the majority of deltamethrin resistance observed in the QTC279 strain of Tribolium castaneum. We used recently completed whole-genome sequence of T. castaneum to prepare custom microarrays and identified a P450 gene, CYP6BQ9, which showed more than a 200-fold higher expression in the deltamethrin-resistant QTC279 strain when compared with its expression in the deltamethrin-susceptible Lab-S strain. Functional studies using both double-strand RNA (dsRNA)-mediated knockdown in the expression of CYP6BQ9 and transgenic expression of CYP6BQ9 in Drosophila melanogaster showed that CYP6BQ9 confers deltamethrin resistance. Furthermore, CYP6BQ9 enzyme expressed in baculovirus metabolizes deltamethrin to 4-hydroxy deltamethrin. Strikingly, we also found that unlike many P450 genes involved in insecticide resistance that were reported previously, CYP6BQ9 is predominantly expressed in the brain, a part of the central nervous system (CNS) containing voltage-gated sodium channels targeted by deltamethrin. Taken together, the current studies on the brain-specific insect P450 involved in deltamethrin resistance shed new light on the understanding of the molecular basis and evolution of insecticide resistance.

  15. Infection of Tribolium castaneum with Bacillus thuringiensis: quantification of bacterial replication within cadavers, transmission via cannibalism, and inhibition of spore germination.

    PubMed

    Milutinović, Barbara; Höfling, Christina; Futo, Momir; Scharsack, Jörn P; Kurtz, Joachim

    2015-12-01

    Reproduction within a host and transmission to the next host are crucial for the virulence and fitness of pathogens. Nevertheless, basic knowledge about such parameters is often missing from the literature, even for well-studied bacteria, such as Bacillus thuringiensis, an endospore-forming insect pathogen, which infects its hosts via the oral route. To characterize bacterial replication success, we made use of an experimental oral infection system for the red flour beetle Tribolium castaneum and developed a flow cytometric assay for the quantification of both spore ingestion by the individual beetle larvae and the resulting spore load after bacterial replication and resporulation within cadavers. On average, spore numbers increased 460-fold, showing that Bacillus thuringiensis grows and replicates successfully in insect cadavers. By inoculating cadaver-derived spores and spores from bacterial stock cultures into nutrient medium, we next investigated outgrowth characteristics of vegetative cells and found that cadaver-derived bacteria showed reduced growth compared to bacteria from the stock cultures. Interestingly, this reduced growth was a consequence of inhibited spore germination, probably originating from the host and resulting in reduced host mortality in subsequent infections by cadaver-derived spores. Nevertheless, we further showed that Bacillus thuringiensis transmission was possible via larval cannibalism when no other food was offered. These results contribute to our understanding of the ecology of Bacillus thuringiensis as an insect pathogen. PMID:26386058

  16. The Expression and Function of the Achaete-Scute Genes in Tribolium castaneum Reveals Conservation and Variation in Neural Pattern Formation and Cell Fate Specification

    NASA Technical Reports Server (NTRS)

    Wheeler, Scott R.; Carrico, Michelle L.; Wilson, Beth A.; Brown, Susan J.; Skeath, James B.

    2003-01-01

    SUMMARY The study of achaete-scute (ac/sc) genes has recently become a paradigm to understand the evolution and development of the arthropod nervous system. We describe the identification and characterization of the ache genes in the coleopteran insect species Tribolium castaneum. We have identified two Tribolium ache genes - achaete-scute homolog (Tc-ASH) a proneural gene and asense (Tc-ase) a neural precursor gene that reside in a gene complex. Focusing on the embryonic central nervous system we fmd that Tc-ASH is expressed in all neural precursors and the proneural clusters from which they segregate. Through RNAi and misexpression studies we show that Tc-ASH is necessary for neural precursor formation in Triboliurn and sufficient for neural precursor formation in Drosophila. Comparison of the function of the Drosophila and Triboliurn proneural ac/sc genes suggests that in the Drosophila lineage these genes have maintained their ancestral function in neural precursor formation and have acquired a new role in the fate specification of individual neural precursors. Furthermore, we find that Tc-use is expressed in all neural precursors suggesting an important and conserved role for asense genes in insect nervous system development. Our analysis of the Triboliurn ache genes indicates significant plasticity in gene number, expression and function, and implicates these modifications in the evolution of arthropod neural development.

  17. The expression and function of the achaete-scute genes in Tribolium castaneum reveals conservation and variation in neural pattern formation and cell fate specification

    NASA Technical Reports Server (NTRS)

    Wheeler, Scott R.; Carrico, Michelle L.; Wilson, Beth A.; Brown, Susan J.; Skeath, James B.

    2003-01-01

    The study of achaete-scute (ac/sc) genes has recently become a paradigm to understand the evolution and development of the arthropod nervous system. We describe the identification and characterization of the ac/sc genes in the coleopteran insect species Tribolium castaneum. We have identified two Tribolium ac/sc genes - achaete-scute homolog (Tc-ASH) a proneural gene and asense (Tc-ase) a neural precursor gene that reside in a gene complex. Focusing on the embryonic central nervous system we find that Tc-ASH is expressed in all neural precursors and the proneural clusters from which they segregate. Through RNAi and misexpression studies we show that Tc-ASH is necessary for neural precursor formation in Tribolium and sufficient for neural precursor formation in Drosophila. Comparison of the function of the Drosophila and Tribolium proneural ac/sc genes suggests that in the Drosophila lineage these genes have maintained their ancestral function in neural precursor formation and have acquired a new role in the fate specification of individual neural precursors. Furthermore, we find that Tc-ase is expressed in all neural precursors suggesting an important and conserved role for asense genes in insect nervous system development. Our analysis of the Tribolium ac/sc genes indicates significant plasticity in gene number, expression and function, and implicates these modifications in the evolution of arthropod neural development.

  18. Infection of Tribolium castaneum with Bacillus thuringiensis: quantification of bacterial replication within cadavers, transmission via cannibalism, and inhibition of spore germination.

    PubMed

    Milutinović, Barbara; Höfling, Christina; Futo, Momir; Scharsack, Jörn P; Kurtz, Joachim

    2015-12-01

    Reproduction within a host and transmission to the next host are crucial for the virulence and fitness of pathogens. Nevertheless, basic knowledge about such parameters is often missing from the literature, even for well-studied bacteria, such as Bacillus thuringiensis, an endospore-forming insect pathogen, which infects its hosts via the oral route. To characterize bacterial replication success, we made use of an experimental oral infection system for the red flour beetle Tribolium castaneum and developed a flow cytometric assay for the quantification of both spore ingestion by the individual beetle larvae and the resulting spore load after bacterial replication and resporulation within cadavers. On average, spore numbers increased 460-fold, showing that Bacillus thuringiensis grows and replicates successfully in insect cadavers. By inoculating cadaver-derived spores and spores from bacterial stock cultures into nutrient medium, we next investigated outgrowth characteristics of vegetative cells and found that cadaver-derived bacteria showed reduced growth compared to bacteria from the stock cultures. Interestingly, this reduced growth was a consequence of inhibited spore germination, probably originating from the host and resulting in reduced host mortality in subsequent infections by cadaver-derived spores. Nevertheless, we further showed that Bacillus thuringiensis transmission was possible via larval cannibalism when no other food was offered. These results contribute to our understanding of the ecology of Bacillus thuringiensis as an insect pathogen.

  19. Evaluation of Structural Treatment Efficacy against Tribolium castaneum and Tribolium confusum (Coleoptera: Tenebrionidae) Using Meta-Analysis of Multiple Studies Conducted in Food Facilities.

    PubMed

    Campbell, James F; Buckman, Karrie A; Fields, Paul G; Subramanyam, Bhadriraju

    2015-10-01

    The phase out of methyl bromide for the treatment of structures where grain is milled or processed has triggered a need to evaluate the effectiveness of alternative structural treatments such as sulfuryl fluoride and heat. Tribolium castaneum (Herbst) (red flour beetle) and Tribolium confusum Jacquelin du Val (confused flour beetle) (Coleoptera: Tenebrionidae) are primary targets for structural treatments, and impact of treatments on these species was evaluated in this study. Efficacy was measured by comparing beetle captures in pheromone- and kairomone-baited traps immediately before and immediately after treatments. Studies were conducted primarily in commercial wheat mills and rice mills, with the treatments conducted by commercial applicators. A meta-analysis approach was used to evaluate efficacy data collected from 111 treatments applied in 39 facilities. Findings confirm that structural fumigations and heat treatments greatly reduce pest populations within food facilities, but there was significant variation in the efficacy of individual treatments. Generally, the different treatment types (sulfuryl fluoride, methyl bromide, and heat) provided similar reductions in beetle capture using multiple metrics of beetle activity. The novel application of meta-analysis to structural treatment efficacy assessment generated a robust estimate of overall treatment efficacy, provided insights into factors potentially impacting efficacy, and identified data gaps that need further research. PMID:26453702

  20. Residual efficacy of methoprene for control of Tribolium castaneum (Coleoptera: Tenebrionidae) larvae at different temperatures on varnished wood, concrete, and wheat.

    PubMed

    Wijayaratne, L K Wolly; Fields, Paul G; Arthur, Frank H

    2012-04-01

    The residual efficacy of the juvenile hormone analog methoprene (Diacon II) was evaluated in bioassays using larvae of Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae) exposed on unsealed concrete or varnished wood treated with a liquid formulation and held at different temperatures. When these two types of surfaces were stored at 20, 30 or 35 degrees C for 0-24 wk, the percentage of adult emergence on concrete increased with time. In contrast, there was no adult emergence from larvae exposed to varnished wood at 24 wk after treatment at any of these temperatures. The presence of flour reduced residual efficacy of methoprene on concrete, but not on varnished wood, with no differences between cleaning frequencies. Methoprene was also stable for 48 h on concrete held at 65 degrees C and wheat, Triticum aestivum L., held at 46 degrees C. Results show that methoprene is stable at a range of temperatures commonly encountered in indoor food storage facilities and at high temperatures attained during insecticidal heat treatments of structures. The residual persistence of methoprene applied to different surface substrates may be affected more by the substrate than by temperature.

  1. Early asymmetries in maternal transcript distribution associated with a cortical microtubule network and a polar body in the beetle Tribolium castaneum.

    PubMed

    Peel, Andrew D; Averof, Michalis

    2010-11-01

    The localization of maternal mRNAs during oogenesis plays a central role in axial specification in some insects. Here we describe a polar body-associated asymmetry in maternal transcript distribution in pre-blastoderm eggs of the beetle Tribolium castaneum. Since the position of the polar body marks the future dorsal side of the embryo, we have investigated whether this asymmetry in mRNA distribution plays a role in dorsal-ventral axis specification. Whilst our results suggest polar body-associated transcripts do not play a significant role in specifying the DV axis, at least during early embryogenesis, we do find that the polar body is closely associated with a cortical microtubule network (CMN), which may play a role in the localization of transcripts during oogenesis. Transcripts of the gene T.c.pangolin co-localize with the CMN at the time of their anterior localization during oogenesis and their anterior localization is disrupted by the microtubule-depolymerizing agent colcemid. PMID:20857499

  2. Identification of G protein-coupled receptors required for vitellogenin uptake into the oocytes of the red flour beetle, Tribolium castaneum.

    PubMed

    Bai, Hua; Palli, Subba Reddy

    2016-01-01

    Previous studies suggested that a membrane receptor might be involved in mediating vitellogenin (Vg) uptake and juvenile hormone (JH)-regulated remodeling of follicular epithelium (also called 'patency'). G protein-coupled receptor (GPCR) family is one of the largest membrane receptor protein families and controls many key physiological processes. To investigate the role of GPCRs in insect reproduction and juvenile hormone-regulated Vg uptake, we performed a comprehensive RNA interference (RNAi) screen targeting GPCRs in the red flour beetle, Tribolium castaneum. Out of 112 GPCRs tested, knockdown of 41 GPCRs resulted in a reduction in fecundity. Interestingly, RNAi against two GPCRs (a Rhodopsin-like receptor and a Dopamine D2-like receptor) led to a significant reduction in Vg accumulation in developing oocytes. Functional assays of these two GPCRs showed that JH triggers a dose-dependent inhibition of intracellular cAMP levels in HEK293 cells expressing Tribolium Dopamine D2-like receptor. These data suggest that Dopamine D2-like receptor plays crucial roles in regulating Vg uptake and is a promising candidate membrane receptor mediating JH regulation of patency in the red flour beetle. PMID:27277501

  3. The negative effect of starvation and the positive effect of mild thermal stress on thermal tolerance of the red flour beetle, Tribolium castaneum

    NASA Astrophysics Data System (ADS)

    Scharf, Inon; Wexler, Yonatan; MacMillan, Heath Andrew; Presman, Shira; Simson, Eddie; Rosenstein, Shai

    2016-04-01

    The thermal tolerance of a terrestrial insect species can vary as a result of differences in population origin, developmental stage, age, and sex, as well as via phenotypic plasticity induced in response to changes in the abiotic environment. Here, we studied the effects of both starvation and mild cold and heat shocks on the thermal tolerance of the red flour beetle, Tribolium castaneum. Starvation led to impaired cold tolerance, measured as chill coma recovery time, and this effect, which was stronger in males than females, persisted for longer than 2 days but less than 7 days. Heat tolerance, measured as heat knockdown time, was not affected by starvation. Our results highlight the difficulty faced by insects when encountering multiple stressors simultaneously and indicate physiological trade-offs. Both mild cold and heat shocks led to improved heat tolerance in both sexes. It could be that both mild shocks lead to the expression of heat shock proteins, enhancing heat tolerance in the short run. Cold tolerance was not affected by previous mild cold shock, suggesting that such a cold shock, as a single event, causes little stress and hence elicits only weak physiological reaction. However, previous mild heat stress led to improved cold tolerance but only in males. Our results point to both hardening and cross-tolerance between cold and heat shocks.

  4. The negative effect of starvation and the positive effect of mild thermal stress on thermal tolerance of the red flour beetle, Tribolium castaneum.

    PubMed

    Scharf, Inon; Wexler, Yonatan; MacMillan, Heath Andrew; Presman, Shira; Simson, Eddie; Rosenstein, Shai

    2016-04-01

    The thermal tolerance of a terrestrial insect species can vary as a result of differences in population origin, developmental stage, age, and sex, as well as via phenotypic plasticity induced in response to changes in the abiotic environment. Here, we studied the effects of both starvation and mild cold and heat shocks on the thermal tolerance of the red flour beetle, Tribolium castaneum. Starvation led to impaired cold tolerance, measured as chill coma recovery time, and this effect, which was stronger in males than females, persisted for longer than 2 days but less than 7 days. Heat tolerance, measured as heat knockdown time, was not affected by starvation. Our results highlight the difficulty faced by insects when encountering multiple stressors simultaneously and indicate physiological trade-offs. Both mild cold and heat shocks led to improved heat tolerance in both sexes. It could be that both mild shocks lead to the expression of heat shock proteins, enhancing heat tolerance in the short run. Cold tolerance was not affected by previous mild cold shock, suggesting that such a cold shock, as a single event, causes little stress and hence elicits only weak physiological reaction. However, previous mild heat stress led to improved cold tolerance but only in males. Our results point to both hardening and cross-tolerance between cold and heat shocks.

  5. Genome-wide analysis of tandem repeats in Tribolium castaneum genome reveals abundant and highly dynamic tandem repeat families with satellite DNA features in euchromatic chromosomal arms.

    PubMed

    Pavlek, Martina; Gelfand, Yevgeniy; Plohl, Miroslav; Meštrović, Nevenka

    2015-12-01

    Although satellite DNAs are well-explored components of heterochromatin and centromeres, little is known about emergence, dispersal and possible impact of comparably structured tandem repeats (TRs) on the genome-wide scale. Our bioinformatics analysis of assembled Tribolium castaneum genome disclosed significant contribution of TRs in euchromatic chromosomal arms and clear predominance of satellite DNA-typical 170 bp monomers in arrays of ≥5 repeats. By applying different experimental approaches, we revealed that the nine most prominent TR families Cast1-Cast9 extracted from the assembly comprise ∼4.3% of the entire genome and reside almost exclusively in euchromatic regions. Among them, seven families that build ∼3.9% of the genome are based on ∼170 and ∼340 bp long monomers. Results of phylogenetic analyses of 2500 monomers originating from these families show high-sequence dynamics, evident by extensive exchanges between arrays on non-homologous chromosomes. In addition, our analysis shows that concerted evolution acts more efficiently on longer than on shorter arrays. Efficient genome-wide distribution of nine TR families implies the role of transposition only in expansion of the most dispersed family, and involvement of other mechanisms is anticipated. Despite similarities in sequence features, FISH experiments indicate high-level compartmentalization of centromeric and euchromatic tandem repeats.

  6. Response of Tribolium castaneum to elevated copper concentrations is influenced by history of metal exposure, sex-specific defences, and infection by the parasite Steinernema feltiae.

    PubMed

    Kramarz, Paulina E; Mordarska, Anna; Mroczka, Magdalena

    2014-07-01

    We studied how copper toxicity in the red flour beetle, Tribolium castaneum changed as a result of infection by the entomopathogenic nematode Steinernema feltiae. Measured traits were: respiration, growth and survival, as well as the concentrations of copper within beetle tissues and in its diet. By comparing F1 and F5 generation we were able to answer how long-term metal exposure changed the responses to both copper and the parasite. The beetles did accumulate copper; however, the results indicated that copper concentrations in beetle tissues were affected by nematode infection, the sex of the experimental animals, and the number of generations of exposure. Five generations of exposure to copper resulted in the highest dry body mass of infected beetles of both sexes; additionally, this group also had the lowest copper concentrations in their tissues. The only factor that had a significant effect on respiration was infection by nematodes: infected beetles of both sexes in both generational groups had significantly decreased respiration rates. Survival was lowest in nematode-infected animals of both sexes from both generations, regardless of exposure to copper. Our results confirm that an organism's response to metal pollution is dependent on its health status and sex. We also found that the history of exposure to metal was equally important-we found enhanced resistance to copper intoxication after only five generations of exposure.

  7. Infection of Tribolium castaneum with Bacillus thuringiensis: Quantification of Bacterial Replication within Cadavers, Transmission via Cannibalism, and Inhibition of Spore Germination

    PubMed Central

    Milutinović, Barbara; Höfling, Christina; Futo, Momir; Scharsack, Jörn P.

    2015-01-01

    Reproduction within a host and transmission to the next host are crucial for the virulence and fitness of pathogens. Nevertheless, basic knowledge about such parameters is often missing from the literature, even for well-studied bacteria, such as Bacillus thuringiensis, an endospore-forming insect pathogen, which infects its hosts via the oral route. To characterize bacterial replication success, we made use of an experimental oral infection system for the red flour beetle Tribolium castaneum and developed a flow cytometric assay for the quantification of both spore ingestion by the individual beetle larvae and the resulting spore load after bacterial replication and resporulation within cadavers. On average, spore numbers increased 460-fold, showing that Bacillus thuringiensis grows and replicates successfully in insect cadavers. By inoculating cadaver-derived spores and spores from bacterial stock cultures into nutrient medium, we next investigated outgrowth characteristics of vegetative cells and found that cadaver-derived bacteria showed reduced growth compared to bacteria from the stock cultures. Interestingly, this reduced growth was a consequence of inhibited spore germination, probably originating from the host and resulting in reduced host mortality in subsequent infections by cadaver-derived spores. Nevertheless, we further showed that Bacillus thuringiensis transmission was possible via larval cannibalism when no other food was offered. These results contribute to our understanding of the ecology of Bacillus thuringiensis as an insect pathogen. PMID:26386058

  8. Identification of G protein-coupled receptors required for vitellogenin uptake into the oocytes of the red flour beetle, Tribolium castaneum

    PubMed Central

    Bai, Hua; Palli, Subba Reddy

    2016-01-01

    Previous studies suggested that a membrane receptor might be involved in mediating vitellogenin (Vg) uptake and juvenile hormone (JH)-regulated remodeling of follicular epithelium (also called ‘patency’). G protein-coupled receptor (GPCR) family is one of the largest membrane receptor protein families and controls many key physiological processes. To investigate the role of GPCRs in insect reproduction and juvenile hormone-regulated Vg uptake, we performed a comprehensive RNA interference (RNAi) screen targeting GPCRs in the red flour beetle, Tribolium castaneum. Out of 112 GPCRs tested, knockdown of 41 GPCRs resulted in a reduction in fecundity. Interestingly, RNAi against two GPCRs (a Rhodopsin-like receptor and a Dopamine D2-like receptor) led to a significant reduction in Vg accumulation in developing oocytes. Functional assays of these two GPCRs showed that JH triggers a dose-dependent inhibition of intracellular cAMP levels in HEK293 cells expressing Tribolium Dopamine D2-like receptor. These data suggest that Dopamine D2-like receptor plays crucial roles in regulating Vg uptake and is a promising candidate membrane receptor mediating JH regulation of patency in the red flour beetle. PMID:27277501

  9. Early asymmetries in maternal transcript distribution associated with a cortical microtubule network and a polar body in the beetle Tribolium castaneum.

    PubMed

    Peel, Andrew D; Averof, Michalis

    2010-11-01

    The localization of maternal mRNAs during oogenesis plays a central role in axial specification in some insects. Here we describe a polar body-associated asymmetry in maternal transcript distribution in pre-blastoderm eggs of the beetle Tribolium castaneum. Since the position of the polar body marks the future dorsal side of the embryo, we have investigated whether this asymmetry in mRNA distribution plays a role in dorsal-ventral axis specification. Whilst our results suggest polar body-associated transcripts do not play a significant role in specifying the DV axis, at least during early embryogenesis, we do find that the polar body is closely associated with a cortical microtubule network (CMN), which may play a role in the localization of transcripts during oogenesis. Transcripts of the gene T.c.pangolin co-localize with the CMN at the time of their anterior localization during oogenesis and their anterior localization is disrupted by the microtubule-depolymerizing agent colcemid.

  10. The red flour beetle Tribolium castaneum allows for the convenient determination of fitness and survival as a measure of toxic effects of the food contaminant acrylamide.

    PubMed

    Grünwald, Stefanie; Gurmai, Ana-Maria; Schuierer, Kristina; Boll, Michael; Wenzel, Uwe

    2014-01-01

    Acrylamide  is a toxic ingredient generated as a by-product of the Maillard reaction in starchy foods processed at temperatures above 120°C. Here we used the red flour beetle Tribolium castaneum as a model organism to test the effects of acrylamide on fitness and survival. Beetles were fed on flour spiked with acrylamide between 0.5% and 10% at 32°C over 2 weeks. Fitness of the beetles was tested by measuring the running distance and survival was recorded after 2 weeks of feeding at increased temperatures at 42°C. Both parameters were dose-dependent reduced by acrylamide. Knockdown of gene homologues of ahr, the arylhydrocarbon receptor, and of nrf-2, the nuclear factor erythroid 2-related factor 2, both reduced fitness and survival. Application of 0.5% acrylamide under knockdown of each factor further reduced fitness and survival, suggesting that ahr and nrf-2 are important for an adequate response to the toxicant. RNA-interference for ahr blocked completely the increase in nrf-2 mRNA levels, suggesting that the actions of ahr on acrylamide detoxification are mediated via Nrf-2.

  11. Insecticide activity of essential oils of Mentha longifolia, Pulicaria gnaphalodes and Achillea wilhelmsii against two stored product pests, the flour beetle, Tribolium castaneum, and the cowpea weevil, Callosobruchus maculatus.

    PubMed

    Khani, Abbas; Asghari, Javad

    2012-01-01

    Essential oils extracted from the foliage of Mentha longifolia (L.) (Lamiales: Lamiaceae) and Pulicaria gnaphalodes Ventenat (Asterales: Asteraceae), and flowers of Achillea wilhelmsii C. Koch (Asterales: Asteraceae) were tested in the laboratory for volatile toxicity against two storedproduct insects, the flour beetle, Tribolium castaneum Herbst (Coleoptera: Tenebrionidae) and the cowpea weevil, Callosobruchus maculatus F. (Coleoptera: Bruchidae). The chemical composition of the isolated oils was examined by gas chromatography-mass spectrometry. InM longifolia, the major compounds were piperitenon (43.9%), tripal (14.3%), oxathiane (9.3%), piperiton oxide (5.9%), and d-limonene (4.3%). In P. gnaphalodes, the major compounds were chrysanthenyl acetate (22.38%), 2L -4L-dihydroxy eicosane (18.5%), verbenol (16.59%), dehydroaromadendrene (12.54%), β-pinen (6.43%), and 1,8 cineol (5.6%). In A. wilhelmsii, the major compounds were 1,8 cineole (13.03%), caranol (8.26%), alpha pinene (6%), farnesyl acetate (6%), and p-cymene (6%). C maculatus was more susceptible to the tested plant products than T castaneum. The oils of the three plants displayed the same insecticidal activity against C. maculatus based on LC(50) values (between 1.54µl/L air in P. gnaphalodes, and 2.65 µl/L air in A. wilhelmsii). While the oils of A. wilhelmsii and M. longifolia showed the same strong insecticidal activity against T. castaneum (LC(50) = 10.02 and 13.05 µl/L air, respectively), the oil of P. gnaphalodes revealed poor activity against the insect (LC(50) = 297.9 µl/L air). These results suggested that essential oils from the tested plants could be used as potential control agents for stored-product insects.

  12. Insecticide Activity of Essential Oils of Mentha longifolia, Pulicaria gnaphalodes and Achillea wilhelmsii Against Two Stored Product Pests, the Flour Beetle, Tribolium castaneum, and the Cowpea Weevil, Callosobruchus maculatus

    PubMed Central

    Khani, Abbas; Asghari, Javad

    2012-01-01

    Essential oils extracted from the foliage of Mentha longifolia (L.) (Lamiales: Lamiaceae) and Pulicaria gnaphalodes Ventenat (Asterales: Asteraceae), and flowers of Achillea wilhelmsii C. Koch (Asterales: Asteraceae) were tested in the laboratory for volatile toxicity against two storedproduct insects, the flour beetle, Tribolium castaneum Herbst (Coleoptera: Tenebrionidae) and the cowpea weevil, Callosobruchus maculatus F. (Coleoptera: Bruchidae). The chemical composition of the isolated oils was examined by gas chromatography-mass spectrometry. InM longifolia, the major compounds were piperitenon (43.9%), tripal (14.3%), oxathiane (9.3%), piperiton oxide (5.9%), and d-limonene (4.3%). In P. gnaphalodes, the major compounds were chrysanthenyl acetate (22.38%), 2L -4L-dihydroxy eicosane (18.5%), verbenol (16.59%), dehydroaromadendrene (12.54%), β-pinen (6.43%), and 1,8 cineol (5.6%). In A. wilhelmsii, the major compounds were 1,8 cineole (13.03%), caranol (8.26%), alpha pinene (6%), farnesyl acetate (6%), and p-cymene (6%). C maculatus was more susceptible to the tested plant products than T castaneum. The oils of the three plants displayed the same insecticidal activity against C. maculatus based on LC50 values (between 1.54µl/L air in P. gnaphalodes, and 2.65 µl/L air in A. wilhelmsii). While the oils of A. wilhelmsii and M. longifolia showed the same strong insecticidal activity against T. castaneum (LC50 = 10.02 and 13.05 µl/L air, respectively), the oil of P. gnaphalodes revealed poor activity against the insect (LC50 = 297.9 µl/L air). These results suggested that essential oils from the tested plants could be used as potential control agents for stored-product insects. PMID:23413994

  13. Loss of Tc-arrow and canonical Wnt signaling alters posterior morphology and pair-rule gene expression in the short-germ insect, Tribolium castaneum.

    PubMed

    Bolognesi, Renata; Fischer, Tamara D; Brown, Susan J

    2009-07-01

    Wnt signaling has been implicated in posterior patterning in short-germ insects, including the red flour beetle Tribolium castaneum (Bolognesi et al. Curr Biol 18:1624-1629, 2008b; Angelini and Kaufman Dev Biol 283:409-423, 2005; Miyawaki et al. Mech Dev 121:119-130, 2004). Specifically, depletion of Wnt ligands Tc-Wnt1 and Tc-WntD/8 produces Tribolium embryos lacking abdominal segments. Similar phenotypes are produced by depletion of Tc-porcupine (Tc-porc) or Tc-pangolin (Tc-pan), indicating that the signal is transmitted through the canonical Wnt pathway (Bolognesi et al. Curr Biol 18:1624-1629, 2008b). Here we show that RNAi for the receptor Tc-arrow produced similar truncated phenotypes, providing additional evidence supporting canonical signal transduction. Furthermore, since in Tribolium segments are defined sequentially by a pair-rule gene circuit that, when interrupted, produces truncated phenotypes (Choe et al. Proc Natl Acad Sci U S A 103:6560-6564, 2006), we investigated the relationship between loss of Wnt signaling and this pair-rule gene circuit. After depletion of the receptor Tc-arrow, expression of Tc-Wnt1 was noticeably absent from the growth zone, while Tc-WntD/8 was restricted to a single spot of expression in what remained of the posterior growth zone. The primary pair-rule genes Tc-runt (Tc-run) and Tc-even-skipped (Tc-eve) were expressed normally in the anterior segments, but were reduced to a single spot in the remnants of the posterior growth zone. Thus, expression of pair-rule genes and Tc-WntD/8 are similarly affected by depletion of Wnt signal and disruption of the posterior growth zone. PMID:19705150

  14. Comparative RNA-sequencing profiling reveals novel Delta-class glutathione S-transferases relative genes expression patterns in Tribolium castaneum.

    PubMed

    Chen, Xuhong; Xiong, Wenfeng; Li, Chengjun; Gao, Shanshan; Song, Xiaowen; Wu, Wei; Li, Bin

    2016-11-15

    Glutathione S-transferases (GSTs) are a large group of enzymes having both detoxification roles conferring insecticide resistance and specialist metabolic functions. Tribolium castaneum GST Delta 1 (TcGSTd1) has been found playing crucial role in insecticide resistance and biological processes in insect species. However, the regulatory system of TcGSTd1 has still rarely been known. Comparing the transcriptome profile of RNAi treated larvae (ds-TcGSTd1) and control larvae of T. canstaneum by using RNA-sequencing, we obtained 14,284,085 sequence reads aligned with 13,275 genes. And 512 differentially expressed genes (DEGs) were identified from ds-TcGSTd1 treated group. Est/CCE, CYP, MRPs were significantly down-regulated in ds-TcGSTd1 group when compared with control group, which illustrated that they cooperated with TcGSTd1 to reduce the activity of cellular metabolism system. While, SNO was up-regulated in ds-TcGSTd1 insects suggested it may also involve in detoxifying alkaloid of insect metabolism system. These results established that TcGSTd1 not only acts as a vital gene for phase II cellular detoxification but also participates in phase 0, I, and III cellular detoxification by cooperating with CSPs, OBPs, CYP9, ESTB1, CCE6, MRPs and other detoxification genes. Knockdown of TcGSTd1 also suppressed several genes encoding antioxidant enzymes, e.g. CuZnSOD, Duox, Prx, HPX, CPO, and MCORP. Suggested that they may modulate the function of TcGSTd1 on lifespan, immune, development and reproduction. All these results shed the new insights into the regulatory mechanism of TcGSTd1 involved in insect physiology and could further facilitate the research of suitable and sustainable managements for the pest control. PMID:27511373

  15. Unexpected functional diversity in the fatty acid desaturases of the flour beetle Tribolium castaneum and identification of key residues determining activity.

    PubMed

    Haritos, Victoria S; Horne, Irene; Damcevski, Katherine; Glover, Karen; Gibb, Nerida

    2014-08-01

    Desaturases catalyse modifications to fatty acids which are essential to homeostasis and for pheromone and defensive chemical production. All desaturases of the flour beetle Tribolium castaneum were investigated via query of the sequenced genome which yielded 15 putative acyl-Coenzyme A genes. Eleven desaturase mRNA were obtained in full length and functionally expressed in yeast. Phylogenetic analysis separated the desaturases into 4 distinct clades; one clade contained conserved beetle Δ9 desaturases, second clade was Tribolium-specific having diverse activities including Δ5, Δ9 and Δ12 desaturation and the other 2 clades had mixed insect representatives. Three members of this clade contained unusual inserted sequences of ∼20 residues in the C-terminal region and were related to desaturases that all contained similar inserts. Deletion of the entirety of the insert in the flour beetle Δ12 desaturase abolished its activity but this was partially restored by the reintroduction of two histidine residues, suggesting the histidine(s) are required for activity but the full length insert is not. Five new desaturase activities were discovered: Δ9 desaturation of C12:0-C16:0 substrates; two unprecedented Δ5 enzymes acting on C18:0 and C16:0; Δ9 activity exclusively on C16:0 and a further stearate Δ9 desaturase. qPCR analysis ruled out a role in sex pheromone synthesis for the Δ5 and Δ9/C16:0 desaturases. The flour beetle genome has underpinned an examination of all transcribed desaturases in the organism and revealed a diversity of novel and unusual activities, an improved understanding of the evolutionary relationships among insect desaturases and sequence determinants of activity.

  16. Metamorphic labral axis patterning in the beetle Tribolium castaneum requires multiple upstream, but few downstream, genes in the appendage patterning network.

    PubMed

    Smith, Frank W; Angelini, David R; Gaudio, Matthew S; Jockusch, Elizabeth L

    2014-03-01

    The arthropod labrum is an anterior appendage-like structure that forms the dorsal side of the preoral cavity. Conflicting interpretations of fossil, nervous system, and developmental data have led to a proliferation of scenarios for labral evolution. The best supported hypothesis is that the labrum is a novel structure that shares development with appendages as a result of co-option. Here, we use RNA interference in the red flour beetle Tribolium castaneum to compare metamorphic patterning of the labrum to previously published data on ventral appendage patterning. As expected under the co-option hypothesis, depletion of several genes resulted in similar defects in the labrum and ventral appendages. These include proximal deletions and proximal-to-distal transformations resulting from depletion of the leg gap genes homothorax and extradenticle, large-scale deletions resulting from depletion of the leg gap gene Distal-less, and smaller distal deletions resulting from knockdown of the EGF ligand Keren. However, depletion of dachshund and many of the genes that function downstream of the leg gap genes in the ventral appendages had either subtle or no effects on labral axis patterning. This pattern of partial similarity suggests that upstream genes act through different downstream targets in the labrum. We also discovered that many appendage axis patterning genes have roles in patterning the epipharyngeal sensillum array, suggesting that they have become integrated into a novel regulatory network. These genes include Notch, Delta, and decapentaplegic, and the transcription factors abrupt, bric à brac, homothorax, extradenticle and the paralogs apterous a and apterous b.

  17. Population growth rate and genetic variability of small and large populations of Red flour beetle (Tribolium castaneum) following multigenerational exposure to copper.

    PubMed

    Laskowski, Ryszard; Radwan, Jacek; Kuduk, Katarzyna; Mendrok, Magdalena; Kramarz, Paulina

    2015-07-01

    We reared large (1000 individuals) and small (20 individuals) populations of Tribolium castaneum on diet contaminated with copper in order to determine if the size of a population affects its ability to adapt to adverse environmental conditions. After 10 generations, we used microsatellite markers to estimate and subsequently compare the genetic variability of the copper-treated populations with that of the control populations, which were reared on uncontaminated medium. Additionally, we conducted a full cross-factorial experiment which evaluated the effects of 10 generations of "pre-exposure" to copper on a population's fitness in control and copper-contaminated environments. In order to distinguish results potentially arising from genetic adaptation from those due to non-genetic effects associated to parental exposure to copper, we subjected also F11 generation, originating from parents not exposed to copper, to the same cross-factorial experiment. The effects of long-term exposure to copper depended on population size: the growth rates of small populations that were pre-exposed to copper were inhibited compared to those of small populations reared in uncontaminated environments. Large Cu-exposed populations had a higher growth rate in the F10 generation compared to the control groups, while the growth rate of the F11 generation was unaffected by copper exposure history. The only factor that had a significant effect on genetic variability was population size, but this was to be expected given the large difference in the number of individuals between large and small populations. Neither copper contamination nor its interaction with population size affected the number of microsatellite alleles retained in the F10 generation.

  18. Germ-cell cluster formation in the telotrophic meroistic ovary of Tribolium castaneum (Coleoptera, Polyphaga, Tenebrionidae) and its implication on insect phylogeny.

    PubMed

    Trauner, Jochen; Büning, Jürgen

    2007-01-01

    Tribolium castaneum has telotrophic meroistic ovarioles of the Polyphaga type. During larval stages, germ cells multiply in a first mitotic cycle forming many small, irregularly branched germ-cell clusters which colonize between the anterior and posterior somatic tissues in each ovariole. Because germ-cell multiplication is accompanied by cluster splitting, we assume a very low number of germ cells per ovariole at the beginning of ovariole development. In the late larval and early pupal stages, we found programmed cell death of germ-cell clusters that are located in anterior and middle regions of the ovarioles. Only those clusters survive that rest on posterior somatic tissue. The germ cells that are in direct contact with posterior somatic cells transform into morphologically distinct pro-oocytes. Intercellular bridges interconnecting pro-oocytes are located posteriorly and are filled with fusomes that regularly fuse to form polyfusomes. Intercellular bridges connecting pro-oocytes to pro-nurse cells are always positioned anteriorly and contain small fusomal plugs. During pupal stages, a second wave of metasynchronous mitoses is initiated by the pro-oocytes, leading to linear subclusters with few bifurcations. We assume that the pro-oocytes together with posterior somatic cells build the center of determination and differentiation of germ cells throughout the larval, pupal, and adult stages. The early developmental pattern of germ-cell multiplication is highly similar to the events known from the telotrophic ovary of the Sialis type. We conclude that among the common ancestors of Neuropterida and Coleoptera, a telotrophic meroistic ovary of the Sialis type evolved, which still exists in Sialidae, Raphidioptera, and a myxophagan Coleoptera family, the Hydroscaphidae. Consequently, the telotrophic ovary of the Polyphaga type evolved from the Sialis type.

  19. Loss of Tc-arrow and canonical Wnt signaling alters posterior morphology and pair-rule gene expression in the short-germ insect, Tribolium castaneum.

    PubMed

    Bolognesi, Renata; Fischer, Tamara D; Brown, Susan J

    2009-07-01

    Wnt signaling has been implicated in posterior patterning in short-germ insects, including the red flour beetle Tribolium castaneum (Bolognesi et al. Curr Biol 18:1624-1629, 2008b; Angelini and Kaufman Dev Biol 283:409-423, 2005; Miyawaki et al. Mech Dev 121:119-130, 2004). Specifically, depletion of Wnt ligands Tc-Wnt1 and Tc-WntD/8 produces Tribolium embryos lacking abdominal segments. Similar phenotypes are produced by depletion of Tc-porcupine (Tc-porc) or Tc-pangolin (Tc-pan), indicating that the signal is transmitted through the canonical Wnt pathway (Bolognesi et al. Curr Biol 18:1624-1629, 2008b). Here we show that RNAi for the receptor Tc-arrow produced similar truncated phenotypes, providing additional evidence supporting canonical signal transduction. Furthermore, since in Tribolium segments are defined sequentially by a pair-rule gene circuit that, when interrupted, produces truncated phenotypes (Choe et al. Proc Natl Acad Sci U S A 103:6560-6564, 2006), we investigated the relationship between loss of Wnt signaling and this pair-rule gene circuit. After depletion of the receptor Tc-arrow, expression of Tc-Wnt1 was noticeably absent from the growth zone, while Tc-WntD/8 was restricted to a single spot of expression in what remained of the posterior growth zone. The primary pair-rule genes Tc-runt (Tc-run) and Tc-even-skipped (Tc-eve) were expressed normally in the anterior segments, but were reduced to a single spot in the remnants of the posterior growth zone. Thus, expression of pair-rule genes and Tc-WntD/8 are similarly affected by depletion of Wnt signal and disruption of the posterior growth zone.

  20. Gene Expression Levels Are Correlated with Synonymous Codon Usage, Amino Acid Composition, and Gene Architecture in the Red Flour Beetle, Tribolium castaneum

    PubMed Central

    Williford, Anna; Demuth, Jeffery P.

    2012-01-01

    Gene expression levels correlate with multiple aspects of gene sequence and gene structure in phylogenetically diverse taxa, suggesting an important role of gene expression levels in the evolution of protein-coding genes. Here we present results of a genome-wide study of the influence of gene expression on synonymous codon usage, amino acid composition, and gene structure in the red flour beetle, Tribolium castaneum. Consistent with the action of translational selection, we find that synonymous codon usage bias increases with gene expression. However, the correspondence between tRNA gene copy number and optimal codons is weak. At the amino acid level, translational selection is suggested by the positive correlation between tRNA gene numbers and amino acid usage, which is stronger for highly expressed genes. In addition, there is a clear trend for increased use of metabolically cheaper, less complex amino acids as gene expression increases. tRNA gene numbers also correlate negatively with amino acid size/complexity (S/C) score indicating the coupling between translational selection and selection to minimize the use of large/complex amino acids. Interestingly, the analysis of 10 additional genomes suggests that the correlation between tRNA gene numbers and amino acid S/C score is widespread and might be explained by selection against negative consequences of protein misfolding. At the level of gene structure, three major trends are detected: 1) complete coding region length increases across low and intermediate expression levels but decreases in highly expressed genes; 2) the average intron size shows the opposite trend, first decreasing with expression, followed by a slight increase in highly expressed genes; and 3) intron density remains nearly constant across all expression levels. These changes in gene architecture are only in partial agreement with selection favoring reduced cost of biosynthesis. PMID:22826459

  1. Metamorphic labral axis patterning in the beetle Tribolium castaneum requires multiple upstream, but few downstream, genes in the appendage patterning network

    PubMed Central

    Smith, Frank W.; Angelini, David R.; Gaudio, Matthew; Jockusch, Elizabeth L.

    2014-01-01

    The arthropod labrum is an anterior appendage-like structure that forms the dorsal side of the preoral cavity. Conflicting interpretations of fossil, nervous system and developmental data have led to a proliferation of scenarios for labral evolution. The best supported hypothesis is that the labrum is a novel structure that shares development with appendages as a result of co-option. Here, we use RNA interference in the red flour beetle Tribolium castaneum to compare metamorphic patterning of the labrum to previously published data on ventral appendage patterning. As expected under the co-option hypothesis, depletion of several genes resulted in similar defects in the labrum and ventral appendages. These include proximal deletions and proximal-to-distal transformations resulting from depletion of the leg gap genes homothorax and extradenticle, large-scale deletions resulting from depletion of the leg gap gene Distal-less, and smaller distal deletions resulting from knockdown of the EGF ligand Keren. However, depletion of dachshund and many of the genes that function downstream of the leg gap genes in the ventral appendages had either subtle or no effects on labral axis patterning. This pattern of partial similarity suggests that upstream genes act through different downstream targets in the labrum. We also discovered that many appendage axis patterning genes have roles in patterning the epipharyngeal sensillum array, suggesting that they have become integrated into a novel regulatory network. These genes include Notch, Delta, and decapentaplegic, and the transcription factors abrupt, bric à brac, homothorax, extradenticle and the paralogs apterous a and apterous b. PMID:24617987

  2. DIRS retroelements in arthropods: identification of the recently active TcDirs1 element in the red flour beetle Tribolium castaneum.

    PubMed

    Goodwin, T J D; Poulter, R T M; Lorenzen, M D; Beeman, R W

    2004-08-01

    Members of the DIRS family of retrotransposons differ from most other known retrotransposons in that they encode a tyrosine recombinase (YR), a type of enzyme frequently involved in site-specific recombination. This enzyme is believed to insert the extrachromosomal DNA intermediate of DIRS element retrotransposition into the host genome. DIRS elements have been found in plants, a slime mold, fungi, and a variety of animals including vertebrates, echinoderms and nematodes. They have a somewhat patchy distribution, however, apparently being absent from a number of model organisms such as Saccharomyces cerevisiae, Arabidopsis thaliana and Drosophila melanogaster. In this report we describe the first DIRS retroelement to be identified in an arthropod. This element, TcDirs1, was found in the red flour beetle Tribolium castaneum (Coleoptera). It is generally similar in sequence and structure to several previously described members of the DIRS group: it is bordered by inverted terminal repeats and it has a similar set of protein-coding domains (Gag, reverse transcriptase/ribonuclease H, and the YR), although these are arranged in a novel fashion. TcDirs1 elements exhibit several features indicative of recent activity, such as intact coding regions, a high level of sequence similarity between distinct elements and polymorphic insertion sites. Given their presence in an experimentally tractable host, these potentially active elements might serve as useful models for the study of DIRS element retrotransposition. An element closely related to TcDirs1 was also detected in sequences from a second arthropod, the honey bee Apis mellifera (Hymenoptera), suggesting that these retrotransposons are long-term residents of arthropod genomes. PMID:15221458

  3. Functional characterization of five different PRXamide receptors of the red flour beetle Tribolium castaneum with peptidomimetics and identification of agonists and antagonists

    PubMed Central

    Jiang, Hongbo; Wei, Zhaojun; Nachman, Ronald J.; Kaczmarek, Krzysztof; Zabrocki, Janusz; Park, Yoonseong

    2014-01-01

    The neuropeptidergic system in insects is an excellent target for pest control strategies. One promising biorational approach is the use of peptidomimetics modified from endogenous ligands to enhance biostability and bioavailability. In this study, we functionally characterized five different G protein-coupled receptors in a phylogenetic cluster, containing receptors for PRXamide in the red flour beetle Tribolium castaneum, by evaluating a series of 70 different peptides and peptidomimetics. Three pyrokinin receptors (TcPKr-A, -B, and –C), cardioacceleratory peptide receptor (TcCAPAr) and ecdysis triggering hormone receptor (TcETHr) were included in the study. Strong agonistic or antagonistic peptidomimetics were identified, and included beta-proline (β3P) modification of the core amino acid residue proline and also a cyclo-peptide. It is common for a ligand to act on multiple receptors. In a number of cases, a ligand acting as an agonist on one receptor was an efficient antagonist on another receptor, suggesting complex outcomes of a peptidomimetic in a biological system. Interestingly, TcPK-A was highly promiscuous with a high number of agonists, while TcPK-C and TcCAPAr had a lower number of agonists, but a higher number of compounds acting as an antagonist. This observation suggests that a target GPCR with more promiscuity will provide better success for peptidomimetic approaches. This study is the first description of peptidomimetics on a CAPA receptor and resulted in the identification of peptidomimetic analogs that demonstrate antagonism of CAPA ligands. The PRXamide receptor assays with peptidomimetics provide useful insights into the biochemical properties of receptors. PMID:25447413

  4. EMS Student Handbook.

    ERIC Educational Resources Information Center

    Ogle, Patrick

    This student guide is one of a series of self-contained materials for students enrolled in an emergency medical services (EMS) training program. Discussed in the individual sections of the guide are the following topics: the purpose and history of EMS professionals; EMS training, certification and examinations (national and state certification and…

  5. EM International. Volume 1

    SciTech Connect

    Not Available

    1993-07-01

    It is the intent of EM International to describe the Office of Environmental Restoration and Waste Management`s (EM`s) various roles and responsibilities within the international community. Cooperative agreements and programs, descriptions of projects and technologies, and synopses of visits to international sites are all highlighted in this semiannual journal. Focus on EM programs in this issue is on international collaboration in vitrification projects. Technology highlights covers: in situ sealing for contaminated sites; and remote sensors for toxic pollutants. Section on profiles of countries includes: Arctic contamination by the former Soviet Union, and EM activities with Germany--cooperative arrangements.

  6. Development and application of a method for analysis of lufenuron in wheat flour by gas chromatography-mass spectrometry and confirmation of bio-efficacy against Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae).

    PubMed

    Ahire, Kedar C; Arora, Manjit S; Mukherjee, Samindra N

    2008-01-01

    A new analytical method using gas chromatography with mass spectrometry (GC-MS) for the quantitative determination of lufenuron, a benzoylphenylurea (BPU) class of insecticide, from wheat flour has been developed and applied for time-dependent residue monitoring in treated wheat flour. The analyte was extracted from wheat flour by a single step solid-liquid extraction by using ethyl acetate and subsequently cleaned up using the Primary Secondary Amine as a sorbent prior to GC-MS analysis. The present method provides sufficient sensitivity as reflected by the values of limit of detection (LOD) and limit of quantification (LOQ), 5 ng/mL (S/N approximately 3) and 50 ng/mL (the lowest validation point on the calibration curve), respectively. The calibration curve showed an excellent linearity in the concentration range of 50-1000 ng/mL (r2=0.998). The average recovery for spiked samples at three concentrations (150, 300, and 450 ng/g) was 98.23+/-2.52% R.S.D. The method was applied for the determination of lufenuron residues in treated wheat flour samples. Simultaneous determination of bio-efficacy of lufenuron residues was also carried out against the red flour beetle, Tribolium castaneum to correlate the actual residual effect of lufenuron as detected by the analytical method, over a period of 3 months. The findings revealed that the residual concentration of lufenuron were neither uniform nor in descending order over a period of 3 months in wheat flour, possibly because of an uneven dispersal in the treated wheat which was subsequently milled into flour, as confirmed by GC-MS analysis. However, the residues of lufenuron were sufficient to produce 100% mortality of T. castaneum larvae up to 3 months. The results have been discussed in view of the potential of lufenuron as a candidate molecule for the control of stored product pests. PMID:18078793

  7. EMS in Mauritius.

    PubMed

    Ramalanjaona, Georges; Brogan, Gerald X

    2009-02-01

    Mauritius lies in the southwest Indian Ocean about 1250 miles from the African coast and 500 miles from Madagascar. Mauritius (estimated population 1,230,602) became independent from the United Kingdom in 1968 and has one of the highest GDP per capita in Africa. Within Mauritius there is a well established EMS system with a single 999 national dispatch system. Ambulances are either publicly or privately owned. Public ambulances are run by the Government (SAMU). Megacare is a private subscriber only ambulance service. The Government has recently invested in new technology such as telemedicine to further enhance the role of EMS on the island. This article describes the current state of EMS in Mauritius and depicts its development in the context of Government effort to decentralise and modernise the healthcare system.

  8. The EM Earthquake Precursor

    NASA Astrophysics Data System (ADS)

    Jones, K. B., II; Saxton, P. T.

    2013-12-01

    Many attempts have been made to determine a sound forecasting method regarding earthquakes and warn the public in turn. Presently, the animal kingdom leads the precursor list alluding to a transmission related source. By applying the animal-based model to an electromagnetic (EM) wave model, various hypotheses were formed, but the most interesting one required the use of a magnetometer with a differing design and geometry. To date, numerous, high-end magnetometers have been in use in close proximity to fault zones for potential earthquake forecasting; however, something is still amiss. The problem still resides with what exactly is forecastable and the investigating direction of EM. After the 1989 Loma Prieta Earthquake, American earthquake investigators predetermined magnetometer use and a minimum earthquake magnitude necessary for EM detection. This action was set in motion, due to the extensive damage incurred and public outrage concerning earthquake forecasting; however, the magnetometers employed, grounded or buried, are completely subject to static and electric fields and have yet to correlate to an identifiable precursor. Secondly, there is neither a networked array for finding any epicentral locations, nor have there been any attempts to find even one. This methodology needs dismissal, because it is overly complicated, subject to continuous change, and provides no response time. As for the minimum magnitude threshold, which was set at M5, this is simply higher than what modern technological advances have gained. Detection can now be achieved at approximately M1, which greatly improves forecasting chances. A propagating precursor has now been detected in both the field and laboratory. Field antenna testing conducted outside the NE Texas town of Timpson in February, 2013, detected three strong EM sources along with numerous weaker signals. The antenna had mobility, and observations were noted for recurrence, duration, and frequency response. Next, two

  9. Identified EM Earthquake Precursors

    NASA Astrophysics Data System (ADS)

    Jones, Kenneth, II; Saxton, Patrick

    2014-05-01

    Many attempts have been made to determine a sound forecasting method regarding earthquakes and warn the public in turn. Presently, the animal kingdom leads the precursor list alluding to a transmission related source. By applying the animal-based model to an electromagnetic (EM) wave model, various hypotheses were formed, but the most interesting one required the use of a magnetometer with a differing design and geometry. To date, numerous, high-end magnetometers have been in use in close proximity to fault zones for potential earthquake forecasting; however, something is still amiss. The problem still resides with what exactly is forecastable and the investigating direction of EM. After a number of custom rock experiments, two hypotheses were formed which could answer the EM wave model. The first hypothesis concerned a sufficient and continuous electron movement either by surface or penetrative flow, and the second regarded a novel approach to radio transmission. Electron flow along fracture surfaces was determined to be inadequate in creating strong EM fields, because rock has a very high electrical resistance making it a high quality insulator. Penetrative flow could not be corroborated as well, because it was discovered that rock was absorbing and confining electrons to a very thin skin depth. Radio wave transmission and detection worked with every single test administered. This hypothesis was reviewed for propagating, long-wave generation with sufficient amplitude, and the capability of penetrating solid rock. Additionally, fracture spaces, either air or ion-filled, can facilitate this concept from great depths and allow for surficial detection. A few propagating precursor signals have been detected in the field occurring with associated phases using custom-built loop antennae. Field testing was conducted in Southern California from 2006-2011, and outside the NE Texas town of Timpson in February, 2013. The antennae have mobility and observations were noted for

  10. EPA LABORATORIES IMPLEMENT EMS PROGRAM

    EPA Science Inventory

    This paper highlights the breadth and magnitude of carrying out an effective Environmental Management System (EMS) program at the U.S. EPA's research and development laboratories. Federal research laboratories have unique operating challenges compared to more centralized industr...

  11. Busca de estruturas em grandes escalas em altos redshifts

    NASA Astrophysics Data System (ADS)

    Boris, N. V.; Sodrã©, L., Jr.; Cypriano, E.

    2003-08-01

    A busca por estruturas em grandes escalas (aglomerados de galáxias, por exemplo) é um ativo tópico de pesquisas hoje em dia, pois a detecção de um único aglomerado em altos redshifts pode por vínculos fortes sobre os modelos cosmológicos. Neste projeto estamos fazendo uma busca de estruturas distantes em campos contendo pares de quasares próximos entre si em z Â3 0.9. Os pares de quasares foram extraídos do catálogo de Véron-Cetty & Véron (2001) e estão sendo observados com os telescópios: 2,2m da University of Hawaii (UH), 2,5m do Observatório de Las Campanas e com o GEMINI. Apresentamos aqui a análise preliminar de um par de quasares observado nos filtros i'(7800 Å) e z'(9500 Å) com o GEMINI. A cor (i'-z') mostrou-se útil para detectar objetos "early-type" em redshifts menores que 1.1. No estudo do par 131046+0006/J131055+0008, com redshift ~ 0.9, o uso deste método possibilitou a detecção de sete objetos candidatos a galáxias "early-type". Num mapa da distribuição projetada dos objetos para 22 < i' < 25 observou-se que estas galáxias estão localizadas próximas a um dos quasares e há indícios de que estejam aglomeradas dentro de um área de ~ 6 arcmin2. Se esse for o caso, estes objetos seriam membros de uma estrutura em grande escala. Um outro argumento em favor dessa hipótese é que eles obedecem uma relação do tipo Kormendy (raio equivalente X brilho superficial dentro desse raio), como a apresentada pelas galáxias elípticas em z = 0.

  12. The National EMS Research strategic plan.

    PubMed

    Sayre, Michael R; White, Lynn J; Brown, Lawrence H; McHenry, Susan D

    2005-01-01

    One of the eight major recommendations put forth by the National EMS Research Agenda Implementation Project in 2002 was the development of an emergency medical services (EMS) research strategic plan. Using a modified Delphi technique along with a consensus conference approach, a strategic plan for EMS research was created. The plan includes recommendations for concentrating efforts by EMS researchers, policy makers, and funding resources with the ultimate goal of improving clinical outcomes. Clinical issues targeted for additional research efforts include evaluation and treatment of patients with asthma, acute cardiac ischemia, circulatory shock, major injury, pain, acute stroke, and traumatic brain injury. The plan calls for developing, evaluating, and validating improved measurement tools and techniques. Additional research to improve the education of EMS personnel as well as system design and operation is also suggested. Implementation of the EMS research strategic plan will improve both the delivery of services and the care of individuals who access the emergency medical system.

  13. DOE/EM Criticality Safety Needs Assessment

    SciTech Connect

    Westfall, Robert Michael; Hopper, Calvin Mitchell

    2011-02-01

    The issue of nuclear criticality safety (NCS) in Department of Energy Environmental Management (DOE/EM) fissionable material operations presents challenges because of the large quantities of material present in the facilities and equipment that are committed to storage and/or material conditioning and dispositioning processes. Given the uncertainty associated with the material and conditions for many DOE/EM fissionable material operations, ensuring safety while maintaining operational efficiency requires the application of the most-effective criticality safety practices. In turn, more-efficient implementation of these practices can be achieved if the best NCS technologies are utilized. In 2002, DOE/EM-1 commissioned a survey of criticality safety technical needs at the major EM sites. These needs were documented in the report Analysis of Nuclear Criticality Safety Technology Supporting the Environmental Management Program, issued May 2002. Subsequent to this study, EM safety management personnel made a commitment to applying the best and latest criticality safety technology, as described by the DOE Nuclear Criticality Safety Program (NCSP). Over the past 7 years, this commitment has enabled the transfer of several new technologies to EM operations. In 2008, it was decided to broaden the basis of the EM NCS needs assessment to include not only current needs for technologies but also NCS operational areas with potential for improvements in controls, analysis, and regulations. A series of NCS workshops has been conducted over the past years, and needs have been identified and addressed by EM staff and contractor personnel. These workshops were organized and conducted by the EM Criticality Safety Program Manager with administrative and technical support by staff at Oak Ridge National Laboratory (ORNL). This report records the progress made in identifying the needs, determining the approaches for addressing these needs, and assimilating new NCS technologies into EM

  14. EM international activities. February 1997 highlights

    SciTech Connect

    1997-02-01

    EM International Highlights is a brief summary of on-going international projects within the Department of Energy`s Office of Environmental Management (EM). This document contains sections on: Global Issues, activities in Western Europe, activities in central and Eastern Europe, activities in Russia, activities in Asia and the Pacific Rim, activities in South America, activities in North America, and International Organizations.

  15. School Budget Hold'em Facilitator's Guide

    ERIC Educational Resources Information Center

    Education Resource Strategies, 2012

    2012-01-01

    "School Budget Hold'em" is a game designed to help school districts rethink their budgeting process. It evolved out of Education Resource Strategies' (ERS) experience working with large urban districts around the country. "School Budget Hold'em" offers a completely new approach--one that can turn the budgeting process into a long-term visioning…

  16. EMS: part three. Preventive medicine for EMS maladies

    SciTech Connect

    Hyfantis, J.F.

    1983-05-01

    Shakeout problems in the fast-growing field of energy-management systems (EMS) are gorwing pains and neither unexpected nor insoluble. A National Bureau of Standards (NBS) survey found user complaints ranging from problems with food spoilage to poorly trained service people. But blame can be placed on users, manufacturers, and distributors. Involving operators and managers in load-control strategies can alleviate some problems and save some money. Allowing for realistic testing time in the installation phase can also save in the long run, as will initiating a maintenance schedule and maintaining operating logs. Software maintenance can present a serious problem. Another NBS study result indicates there is perceptible improvement in system performance when good training accompanies installation, particularly when the training is provided at the user facility. User education is probably the best key to avoiding most of the potential pitfalls. 2 figures, 2 tables.

  17. EM International, July 1994, Volume 2

    SciTech Connect

    Not Available

    1994-10-01

    The Office of Environmental Management (EM) at the Department of Energy (DOE) is seeking out and leveraging foreign technology, data, and resources in keeping with EM`s mandate to protect public health and the environment through the safe and cost-effective remediation of the Department`s nuclear weapons sites. EM works closely with foreign governments, industry, and universities to obtain innovative environmental technologies, scientific and engineering expertise, and operations experience that will support EM`s objectives. Where appropriate, these international resources are used to manage the more urgent risks at our sites, secure a safe workplace, help build consensus on critical issues, and strengthen our technology development program. Through international agreements EM engages in cooperative exchange of information, technology, and individuals. Currently, we are managing agreements with a dozen countries in Europe, Latin America, and Asia. These agreements focus on environmental restoration, waste management, transportation of radioactive wastes, and decontamination and decommissioning. This publication contains the following articles: in situ remediation integrated program; in-situ characterization and inspection of tanks; multimedia environmental pollutant assessment system (MEPAS); LLNL wet oxidation -- AEA technology. Besides these articles, this publication covers: EU activities with Russia; technology transfer activities; and international organization activities.

  18. Project X RFQ EM Design

    SciTech Connect

    Romanov, Gennady; Hoff, Matthew; Li, Derun; Staples, John; Virostek, Steve; /LBNL

    2012-05-09

    Project X is a proposed multi-MW proton facility at Fermi National Accelerator Laboratory (FNAL). The Project X front-end would consist of an H- ion source, a low-energy beam transport (LEBT), a CW 162.5 MHz radio-frequency quadrupole (RFQ) accelerator, and a medium-energy beam transport (MEBT). Lawrence Berkeley National Laboratory (LBNL) and FNAL collaboration is currently developing the designs for various components in the Project X front end. This paper reports the detailed EM design of the CW 162.5 MHz RFQ that provides bunching of the 1-10 mA H- beam with acceleration from 30 keV to 2.1 MeV.

  19. Unified Data Resource for CryoEM

    PubMed Central

    Lawson, Catherine L.

    2010-01-01

    3D cryo-electron microscopy reconstruction methods are uniquely able to reveal structures of many important macromolecules and macromolecular complexes. EMDataBank.org, a joint effort of the Protein Data Bank in Europe (PDBe), the Research Collaboratory for Structural Bioinformatics (RCSB), and the National Center for Macromolecular Imaging (NCMI), is a “one-stop shop” resource for global deposition and retrieval of cryoEM map, model and associated metadata. The resource unifies public access to the two major EM Structural Data archives: EM Data Bank (EMDB) and Protein Data Bank (PDB), and facilitates use of EM structural data of macromolecules and macromolecular complexes by the wider scientific community. PMID:20888470

  20. EMS offshore. A new horizon for paramedics.

    PubMed

    Mallard, A S

    1991-10-01

    The difficulty in getting medical aid to offshore drilling platforms can be a source of life-threatening delays. Recently, some companies have charted new waters by actually stationing EMS crews on their rigs.

  1. EMS offshore. A new horizon for paramedics.

    PubMed

    Mallard, A S

    1991-10-01

    The difficulty in getting medical aid to offshore drilling platforms can be a source of life-threatening delays. Recently, some companies have charted new waters by actually stationing EMS crews on their rigs. PMID:10116023

  2. EMS adaptation for climate change

    NASA Astrophysics Data System (ADS)

    Pan, C.; Chang, Y.; Wen, J.; Tsai, M.

    2010-12-01

    The purpose of this study was to find an appropriate scenario of pre-hospital transportation of an emergency medical service (EMS) system for burdensome casualties resulting from extreme climate events. A case of natural catastrophic events in Taiwan, 88 wind-caused disasters, was reviewed and analyzed. A sequential-conveyance method was designed to shorten the casualty transportation time and to promote the efficiency of ambulance services. A proposed mobile emergency medical center was first constructed in a safe area, but nearby the disaster area. The Center consists of professional medical personnel who process the triage of incoming patients and take care of casualties with minor injuries. Ambulances in the Center were ready to sequentially convey the casualties with severer conditions to an assigned hospital that is distant from the disaster area for further treatment. The study suggests that if we could construct a spacious and well-equipped mobile emergency medical center, only a small portion of casualties would need to be transferred to distant hospitals. This would reduce the over-crowding problem in hospital ERs. First-line ambulances only reciprocated between the mobile emergency medical center and the disaster area, saving time and shortening the working distances. Second-line ambulances were highly regulated between the mobile emergency medical center and requested hospitals. The ambulance service of the sequential-conveyance method was found to be more efficient than the conventional method and was concluded to be more profitable and reasonable on paper in adapting to climate change. Therefore, additional practical work should be launched to collect more precise quantitative data.

  3. 10 CFR Appendixes E-M to Part 52 - [Reserved

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 10 Energy 2 2010-01-01 2010-01-01 false E Appendixes E-M to Part 52 Energy NUCLEAR REGULATORY COMMISSION (CONTINUED) LICENSES, CERTIFICATIONS, AND APPROVALS FOR NUCLEAR POWER PLANTS Appendixes E-M to Part 52...

  4. 10 CFR Appendixes E-M to Part 52 - [Reserved

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 10 Energy 2 2012-01-01 2012-01-01 false E Appendixes E-M to Part 52 Energy NUCLEAR REGULATORY COMMISSION (CONTINUED) LICENSES, CERTIFICATIONS, AND APPROVALS FOR NUCLEAR POWER PLANTS Appendixes E-M to Part 52...

  5. 10 CFR Appendixes E-M to Part 52 - [Reserved

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 10 Energy 2 2014-01-01 2014-01-01 false E Appendixes E-M to Part 52 Energy NUCLEAR REGULATORY COMMISSION (CONTINUED) LICENSES, CERTIFICATIONS, AND APPROVALS FOR NUCLEAR POWER PLANTS Appendixes E-M to Part 52...

  6. 10 CFR Appendixes E-M to Part 52 - [Reserved

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 10 Energy 2 2011-01-01 2011-01-01 false E Appendixes E-M to Part 52 Energy NUCLEAR REGULATORY COMMISSION (CONTINUED) LICENSES, CERTIFICATIONS, AND APPROVALS FOR NUCLEAR POWER PLANTS Appendixes E-M to Part 52...

  7. 10 CFR Appendixes E-M to Part 52 - [Reserved

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 10 Energy 2 2013-01-01 2013-01-01 false E Appendixes E-M to Part 52 Energy NUCLEAR REGULATORY COMMISSION (CONTINUED) LICENSES, CERTIFICATIONS, AND APPROVALS FOR NUCLEAR POWER PLANTS Appendixes E-M to Part 52...

  8. Processing of Cryo-EM Movie Data.

    PubMed

    Ripstein, Z A; Rubinstein, J L

    2016-01-01

    Direct detector device (DDD) cameras dramatically enhance the capabilities of electron cryomicroscopy (cryo-EM) due to their improved detective quantum efficiency (DQE) relative to other detectors. DDDs use semiconductor technology that allows micrographs to be recorded as movies rather than integrated individual exposures. Movies from DDDs improve cryo-EM in another, more surprising, way. DDD movies revealed beam-induced specimen movement as a major source of image degradation and provide a way to partially correct the problem by aligning frames or regions of frames to account for this specimen movement. In this chapter, we use a self-consistent mathematical notation to explain, compare, and contrast several of the most popular existing algorithms for computationally correcting specimen movement in DDD movies. We conclude by discussing future developments in algorithms for processing DDD movies that would extend the capabilities of cryo-EM even further.

  9. Processing of Cryo-EM Movie Data.

    PubMed

    Ripstein, Z A; Rubinstein, J L

    2016-01-01

    Direct detector device (DDD) cameras dramatically enhance the capabilities of electron cryomicroscopy (cryo-EM) due to their improved detective quantum efficiency (DQE) relative to other detectors. DDDs use semiconductor technology that allows micrographs to be recorded as movies rather than integrated individual exposures. Movies from DDDs improve cryo-EM in another, more surprising, way. DDD movies revealed beam-induced specimen movement as a major source of image degradation and provide a way to partially correct the problem by aligning frames or regions of frames to account for this specimen movement. In this chapter, we use a self-consistent mathematical notation to explain, compare, and contrast several of the most popular existing algorithms for computationally correcting specimen movement in DDD movies. We conclude by discussing future developments in algorithms for processing DDD movies that would extend the capabilities of cryo-EM even further. PMID:27572725

  10. Risk Communication Within the EM Program

    SciTech Connect

    Edelson, M.

    2003-02-26

    The U.S. Department of Energy Environmental Management program (EM) conducts the most extensive environmental remediation effort in the world. The annual EM budgets have exceeded $6,000,000,000 for approximately ten years and EM has assumed responsibility for the cleanup of the largest DOE reservations (i.e., at Hanford, Washington, Aiken, South Carolina, and Idaho Falls, Idaho) as well as the facilities at Rocky Flats, Colorado and in Ohio. Each of these sites has areas of extensive radioactive and chemical contamination, numerous surplus facilities that require decontamination and removal, while some have special nuclear material that requires secure storage. The EM program has been criticized for being ineffective (1) and has been repeatedly reorganized to address perceived shortcomings. The most recent reorganization was announced in 2001 to become effective at the beginning of the 2003 Federal Fiscal Year (i.e., October 2002). It was preceded by a ''top to bottom'' review (TTBR) of the program (2) that identified several deficiencies that were to be corrected as a result of the reorganization. One prominent outcome of the TTBR was the identification of ''risk reduction'' as an organizing principle to prioritize the activities of the new EM program. The new program also sought to accelerate progress by identifying a set of critical activities at each site that could be accelerated and result in more rapid site closure, with attendant risk, cost, and schedule benefits. This paper investigates how the new emphasis on risk reduction in the EM program has been communicated to EM stakeholders and regulators. It focuses on the Rocky Flats Environmental Technology Site (RFETS) as a case study and finds that there is little evidence for a new emphasis on risk reduction in EM communications with RFETS stakeholders. Discussions between DOE and RFETS stakeholders often refer to ''risk,'' but the word serves as a placeholder for other concepts. Thus ''risk'' communication

  11. Modelling and design for PM/EM magnetic bearings

    NASA Technical Reports Server (NTRS)

    Pang, D.; Kirk, J. A.; Anand, D. K.; Johnson, R. G.; Zmood, R. B.

    1992-01-01

    A mathematical model of a permanent magnet/electromagnet (PM/EM) radially active bearing is presented. The bearing is represented by both a reluctance model and a stiffness model. The reluctance model analyzes the magnetic circuit of the PM/EM bearings. By combining the two models, the performance of the bearing can be predicted given geometric dimensions, permanent magnet strength, and the parameters of the EM coils. The overall bearing design including the PM and EM design is subject to the performance requirement and physical constraints. A study of these requirements and constraints is discussed. The PM design is based on the required magnetic flux for proper geometric dimensions and magnet strength. The EM design is based on the stability and force slew rate consideration, and dictates the number of turns for the EM coils and the voltage and current of the power amplifier. An overall PM/EM bearing design methodology is proposed and a case study is also demonstrated.

  12. The E-MS Algorithm: Model Selection with Incomplete Data

    PubMed Central

    Jiang, Jiming; Nguyen, Thuan; Rao, J. Sunil

    2014-01-01

    We propose a procedure associated with the idea of the E-M algorithm for model selection in the presence of missing data. The idea extends the concept of parameters to include both the model and the parameters under the model, and thus allows the model to be part of the E-M iterations. We develop the procedure, known as the E-MS algorithm, under the assumption that the class of candidate models is finite. Some special cases of the procedure are considered, including E-MS with the generalized information criteria (GIC), and E-MS with the adaptive fence (AF; Jiang et al. 2008). We prove numerical convergence of the E-MS algorithm as well as consistency in model selection of the limiting model of the E-MS convergence, for E-MS with GIC and E-MS with AF. We study the impact on model selection of different missing data mechanisms. Furthermore, we carry out extensive simulation studies on the finite-sample performance of the E-MS with comparisons to other procedures. The methodology is also illustrated on a real data analysis involving QTL mapping for an agricultural study on barley grains. PMID:26783375

  13. Crosshole EM in steel-cased boreholes

    SciTech Connect

    Wilt, M.; Lee, K.H.; Becker, A.; Spies, B.; Wang, B.

    1996-07-01

    The application of crosshole EM methods through steel well-casing was investigated in theoretical, laboratory and field studies. A numerical code was developed that calculates the attenuation and phase delay of an EM dipole signal propagated through a steel well casing lodged in a homogeneous medium. The code was validated with a scale model and used for sensitivity studies of casing and formation properties. Finally, field measurements were made in an oil field undergoing waterflooding. Our most important findings are that (1) crosshole surveys are feasible using a well pair with one metallic and one non-metallic casing. (2) The casing effect seems be localized within the pipe section that includes the sensor. (3) The effects of the casing can be corrected using simple means and (4) crosshole field data that are sensitive to both formation and casing were acquired in a working environment.

  14. Generation and identification of Arabidopsis EMS mutants.

    PubMed

    Qu, Li-Jia; Qin, Genji

    2014-01-01

    EMS mutant analysis is a routine experiment to identify new players in a specific biological process or signaling pathway using forward genetics. It begins with the generation of mutants by treating Arabidopsis seeds with EMS. A mutant with a phenotype of interest (mpi) is obtained by screening plants of the M2 generation under a specific condition. Once the phenotype of the mpi is confirmed in the next generation, map-based cloning is performed to locate the mpi mutation. During the map-based cloning, mpi plants (Arabidopsis Columbia-0 (Col-0) ecotype background) are first crossed with Arabidopsis Landsberg erecta (Ler) ecotype, and the presence or absence of the phenotype in the F1 hybrids indicates whether the mpi is recessive or dominant. F2 plants with phenotypes similar to the mpi, if the mpi is recessive, or those without the phenotype, if the mpi is dominant, are used as the mapping population. As few as 24 such plants are selected for rough mapping. After finding one marker (MA) linked to the mpi locus or mutant phenotype, more markers near MA are tested to identify recombinants. The recombinants indicate the interval in which the mpi is located. Additional recombinants and molecular markers are then required to narrow down the interval. This is an iterative process of narrowing down the mapping interval until no further recombinants or molecular markers are available. The genes in the mapping interval are then sequenced to look for the mutation. In the last step, the wild-type or mutated gene is cloned to generate binary constructs. Complementation or recapitulation provides the most convincing evidence in determining the mutation that causes the phenotype of the mpi. Here, we describe the procedures for generating mutants with EMS and analyzing EMS mutations by map-based cloning.

  15. Leukocyte Recognition Using EM-Algorithm

    NASA Astrophysics Data System (ADS)

    Colunga, Mario Chirinos; Siordia, Oscar Sánchez; Maybank, Stephen J.

    This document describes a method for classifying images of blood cells. Three different classes of cells are used: Band Neutrophils, Eosinophils and Lymphocytes. The image pattern is projected down to a lower dimensional sub space using PCA; the probability density function for each class is modeled with a Gaussian mixture using the EM-Algorithm. A new cell image is classified using the maximum a posteriori decision rule.

  16. Evaluation of the Ems Estuary ecosystem model

    NASA Astrophysics Data System (ADS)

    Baretta, J. W.; Ruardij, P.

    1987-11-01

    An ecosystem model is used to calculate and summarize carbon budgets within the Ems Estuary, The Netherlands. The similarity between model calculations and field data is established using a validation procedure. Model results show that the seaward boundary concentration for suspended matter is important in determining whether an estuary is an importer or exporter of carbon. Lowered boundary concentrations of suspended matter enhance pelagic primary production, but reduce sedimentation and hence the carbon flux from pelagic to benthic systems.

  17. TandEM: Titan and Enceladus mission

    USGS Publications Warehouse

    Coustenis, A.; Atreya, S.K.; Balint, T.; Brown, R.H.; Dougherty, M.K.; Ferri, F.; Fulchignoni, M.; Gautier, D.; Gowen, R.A.; Griffith, C.A.; Gurvits, L.I.; Jaumann, R.; Langevin, Y.; Leese, M.R.; Lunine, J.I.; McKay, C.P.; Moussas, X.; Muller-Wodarg, I.; Neubauer, F.; Owen, T.C.; Raulin, F.; Sittler, E.C.; Sohl, F.; Sotin, C.; Tobie, G.; Tokano, T.; Turtle, E.P.; Wahlund, J.-E.; Waite, J.H.; Baines, K.H.; Blamont, J.; Coates, A.J.; Dandouras, I.; Krimigis, T.; Lellouch, E.; Lorenz, R.D.; Morse, A.; Porco, C.C.; Hirtzig, M.; Saur, J.; Spilker, T.; Zarnecki, J.C.; Choi, E.; Achilleos, N.; Amils, R.; Annan, P.; Atkinson, D.H.; Benilan, Y.; Bertucci, C.; Bezard, B.; Bjoraker, G.L.; Blanc, M.; Boireau, L.; Bouman, J.; Cabane, M.; Capria, M.T.; Chassefiere, E.; Coll, P.; Combes, M.; Cooper, J.F.; Coradini, A.; Crary, F.; Cravens, T.; Daglis, I.A.; de Angelis, E.; De Bergh, C.; de Pater, I.; Dunford, C.; Durry, G.; Dutuit, O.; Fairbrother, D.; Flasar, F.M.; Fortes, A.D.; Frampton, R.; Fujimoto, M.; Galand, M.; Grasset, O.; Grott, M.; Haltigin, T.; Herique, A.; Hersant, F.; Hussmann, H.; Ip, W.; Johnson, R.; Kallio, E.; Kempf, S.; Knapmeyer, M.; Kofman, W.; Koop, R.; Kostiuk, T.; Krupp, N.; Kuppers, M.; Lammer, H.; Lara, L.-M.; Lavvas, P.; Le, Mouelic S.; Lebonnois, S.; Ledvina, S.; Li, J.; Livengood, T.A.; Lopes, R.M.; Lopez-Moreno, J. -J.; Luz, D.; Mahaffy, P.R.; Mall, U.; Martinez-Frias, J.; Marty, B.; McCord, T.; Salvan, C.M.; Milillo, A.; Mitchell, D.G.; Modolo, R.; Mousis, O.; Nakamura, M.; Neish, C.D.; Nixon, C.A.; Mvondo, D.N.; Orton, G.; Paetzold, M.; Pitman, J.; Pogrebenko, S.; Pollard, W.; Prieto-Ballesteros, O.; Rannou, P.; Reh, K.; Richter, L.; Robb, F.T.; Rodrigo, R.; Rodriguez, S.; Romani, P.; Bermejo, M.R.; Sarris, E.T.; Schenk, P.; Schmitt, B.; Schmitz, N.; Schulze-Makuch, D.; Schwingenschuh, K.; Selig, A.; Sicardy, B.; Soderblom, L.; Spilker, L.J.; Stam, D.; Steele, A.; Stephan, K.; Strobel, D.F.; Szego, K.; Szopa,

    2009-01-01

    TandEM was proposed as an L-class (large) mission in response to ESA's Cosmic Vision 2015-2025 Call, and accepted for further studies, with the goal of exploring Titan and Enceladus. The mission concept is to perform in situ investigations of two worlds tied together by location and properties, whose remarkable natures have been partly revealed by the ongoing Cassini-Huygens mission. These bodies still hold mysteries requiring a complete exploration using a variety of vehicles and instruments. TandEM is an ambitious mission because its targets are two of the most exciting and challenging bodies in the Solar System. It is designed to build on but exceed the scientific and technological accomplishments of the Cassini-Huygens mission, exploring Titan and Enceladus in ways that are not currently possible (full close-up and in situ coverage over long periods of time). In the current mission architecture, TandEM proposes to deliver two medium-sized spacecraft to the Saturnian system. One spacecraft would be an orbiter with a large host of instruments which would perform several Enceladus flybys and deliver penetrators to its surface before going into a dedicated orbit around Titan alone, while the other spacecraft would carry the Titan in situ investigation components, i.e. a hot-air balloon (Montgolfi??re) and possibly several landing probes to be delivered through the atmosphere. ?? Springer Science + Business Media B.V. 2008.

  18. Test beam performance of CDF plug upgrade EM calorimeter

    SciTech Connect

    Fukui, Y.; CDF Upgrade Group

    1998-01-01

    CDF Plug Upgrade(tile-fiber) EM Calorimeter performed resolution of 15%/{radical}E{circle_plus}0.7% with non-linearity less than 1% in a energy range of 5-180 GeV at Fermilab Test Beam. Transverse uniformity of inside-tower-response of the EM Calorimeter was 2.2% with 56 GeV positron, which was reduced to 1.0% with response map correction. We observed 300 photo electron/GeV in the EM Calorimeter. Ratios of EM Calorimeter response to positron beam to that to {sup 137}Cs Source was stable within 1% in the period of 8 months.

  19. 2. HI PAR (ACQUISITION RADAR) TOWER AND ENLISTED MEN (EM) ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    2. HI PAR (ACQUISITION RADAR) TOWER AND ENLISTED MEN (EM) BARRACKS WITH RADAR ATTACHED. - Nike Hercules Missile Battery Summit Site, Battery Control Administration & Barracks Building, Anchorage, Anchorage, AK

  20. Communication - An Effective Tool for Implementing ISO 14001/EMS

    SciTech Connect

    Rachel Damewood; Bowen Huntsman

    2004-04-01

    The Idaho National Engineering and Environmental Laboratory (INEEL) received ISO 14001/EMS certification in June 2002. Communication played an effective role in implementing ISO 14001/EMS at the INEEL. This paper describes communication strategies used during the implementation and certification processes. The INEEL achieved Integrated Safety Management System (ISMS) and Voluntary Protection Program (VPP) Star status in 2001. ISMS implemented a formal process to plan and execute work. VPP facilitated worker involvement by establishing geographic units at various facilities with employee points of contact and management champions. The INEEL Environmental Management System (EMS) was developed to integrate the environmental functional area into its ISMS and VPP. Since the core functions of ISMS, VPP, and EMS are interchangeable, they were easy to integrate. Communication is essential to successfully implement an EMS. (According to ISO 14001 requirements, communication interacts with 12 other elements of the requirements.) We developed communication strategies that integrated ISMS, VPP, and EMS. For example, the ISMS, VPP, and EMS Web sites communicated messages to the work force, such as “VPP emphasizes the people side of doing business, ISMS emphasizes the system side of doing business, and EMS emphasizes the systems to protect the environment; but they all define work, identify and analyze hazards, and mitigate the hazards.” As a result of this integration, the work force supported and implemented the EMS. In addition, the INEEL established a cross-functional communication team to assist with implementing the EMS. The team included members from the Training and Communication organizations, VPP office, Pollution Prevention, Employee and Media Relations, a union representative, facility environmental support, and EMS staff. This crossfunctional team used various communication strategies to promote our EMS to all organization levels and successfully implemented EMS

  1. EM threat analysis for wireless systems.

    SciTech Connect

    Burkholder, R. J. (Ohio State University Electroscience Laboratory); Mariano, Robert J.; Schniter, P. (Ohio State University Electroscience Laboratory); Gupta, I. J. (Ohio State University Electroscience Laboratory)

    2006-06-01

    Modern digital radio systems are complex and must be carefully designed, especially when expected to operate in harsh propagation environments. The ability to accurately predict the effects of propagation on wireless radio performance could lead to more efficient radio designs as well as the ability to perform vulnerability analyses before and after system deployment. In this report, the authors--experts in electromagnetic (EM) modeling and wireless communication theory--describe the construction of a simulation environment that is capable of quantifying the effects of wireless propagation on the performance of digital communication.

  2. 7 CFR 1945.35 - Special EM loan training.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... Designation Staff to the State to assist the State Director in conducting a training meeting(s) with State... 7 Agriculture 13 2010-01-01 2009-01-01 true Special EM loan training. 1945.35 Section 1945.35...) PROGRAM REGULATIONS (CONTINUED) EMERGENCY Disaster Assistance-General § 1945.35 Special EM loan...

  3. 7 CFR 1945.20 - Making EM loans available.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 7 Agriculture 13 2012-01-01 2012-01-01 false Making EM loans available. 1945.20 Section 1945.20...) PROGRAM REGULATIONS (CONTINUED) EMERGENCY Disaster Assistance-General § 1945.20 Making EM loans available... consideration by the Secretary in making determinations under § 1945.6(c)(3) of this subpart. The State...

  4. 7 CFR 1945.20 - Making EM loans available.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 7 Agriculture 13 2011-01-01 2009-01-01 true Making EM loans available. 1945.20 Section 1945.20...) PROGRAM REGULATIONS (CONTINUED) EMERGENCY Disaster Assistance-General § 1945.20 Making EM loans available... consideration by the Secretary in making determinations under § 1945.6(c)(3) of this subpart. The State...

  5. 7 CFR 1945.20 - Making EM loans available.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 13 2010-01-01 2009-01-01 true Making EM loans available. 1945.20 Section 1945.20...) PROGRAM REGULATIONS (CONTINUED) EMERGENCY Disaster Assistance-General § 1945.20 Making EM loans available... consideration by the Secretary in making determinations under § 1945.6(c)(3) of this subpart. The State...

  6. CryoEM at IUCrJ: a new era

    PubMed Central

    Subramaniam, Sriram; Kühlbrandt, Werner; Henderson, Richard

    2016-01-01

    In this overview, we briefly outline recent advances in electron cryomicroscopy (cryoEM) and explain why the journal IUCrJ, published by the International Union of Crystallography, could provide a natural home for publications covering many present and future developments in the cryoEM field. PMID:26870375

  7. 7 CFR 759.6 - EM to be made available.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... designations. When production losses meet the requirements in § 759.5 and the county has been designated as a disaster area for that reason, or when the discretionary exception to production losses for EM under § 759... eligible producers can receive EM loans. (2) Physical loss notification. When only qualifying...

  8. Refinement of atomic models in high resolution EM reconstructions using Flex-EM and local assessment

    PubMed Central

    Joseph, Agnel Praveen; Malhotra, Sony; Burnley, Tom; Wood, Chris; Clare, Daniel K.; Winn, Martyn; Topf, Maya

    2016-01-01

    As the resolutions of Three Dimensional Electron Microscopic reconstructions of biological macromolecules are being improved, there is a need for better fitting and refinement methods at high resolutions and robust approaches for model assessment. Flex-EM/MODELLER has been used for flexible fitting of atomic models in intermediate-to-low resolution density maps of different biological systems. Here, we demonstrate the suitability of the method to successfully refine structures at higher resolutions (2.5–4.5 Å) using both simulated and experimental data, including a newly processed map of Apo-GroEL. A hierarchical refinement protocol was adopted where the rigid body definitions are relaxed and atom displacement steps are reduced progressively at successive stages of refinement. For the assessment of local fit, we used the SMOC (segment-based Manders’ overlap coefficient) score, while the model quality was checked using the Qmean score. Comparison of SMOC profiles at different stages of refinement helped in detecting regions that are poorly fitted. We also show how initial model errors can have significant impact on the goodness-of-fit. Finally, we discuss the implementation of Flex-EM in the CCP-EM software suite. PMID:26988127

  9. Refinement of atomic models in high resolution EM reconstructions using Flex-EM and local assessment.

    PubMed

    Joseph, Agnel Praveen; Malhotra, Sony; Burnley, Tom; Wood, Chris; Clare, Daniel K; Winn, Martyn; Topf, Maya

    2016-05-01

    As the resolutions of Three Dimensional Electron Microscopic reconstructions of biological macromolecules are being improved, there is a need for better fitting and refinement methods at high resolutions and robust approaches for model assessment. Flex-EM/MODELLER has been used for flexible fitting of atomic models in intermediate-to-low resolution density maps of different biological systems. Here, we demonstrate the suitability of the method to successfully refine structures at higher resolutions (2.5-4.5Å) using both simulated and experimental data, including a newly processed map of Apo-GroEL. A hierarchical refinement protocol was adopted where the rigid body definitions are relaxed and atom displacement steps are reduced progressively at successive stages of refinement. For the assessment of local fit, we used the SMOC (segment-based Manders' overlap coefficient) score, while the model quality was checked using the Qmean score. Comparison of SMOC profiles at different stages of refinement helped in detecting regions that are poorly fitted. We also show how initial model errors can have significant impact on the goodness-of-fit. Finally, we discuss the implementation of Flex-EM in the CCP-EM software suite.

  10. Refinement of atomic models in high resolution EM reconstructions using Flex-EM and local assessment.

    PubMed

    Joseph, Agnel Praveen; Malhotra, Sony; Burnley, Tom; Wood, Chris; Clare, Daniel K; Winn, Martyn; Topf, Maya

    2016-05-01

    As the resolutions of Three Dimensional Electron Microscopic reconstructions of biological macromolecules are being improved, there is a need for better fitting and refinement methods at high resolutions and robust approaches for model assessment. Flex-EM/MODELLER has been used for flexible fitting of atomic models in intermediate-to-low resolution density maps of different biological systems. Here, we demonstrate the suitability of the method to successfully refine structures at higher resolutions (2.5-4.5Å) using both simulated and experimental data, including a newly processed map of Apo-GroEL. A hierarchical refinement protocol was adopted where the rigid body definitions are relaxed and atom displacement steps are reduced progressively at successive stages of refinement. For the assessment of local fit, we used the SMOC (segment-based Manders' overlap coefficient) score, while the model quality was checked using the Qmean score. Comparison of SMOC profiles at different stages of refinement helped in detecting regions that are poorly fitted. We also show how initial model errors can have significant impact on the goodness-of-fit. Finally, we discuss the implementation of Flex-EM in the CCP-EM software suite. PMID:26988127

  11. Electromagnetic optimization of EMS-MAGLEV systems

    SciTech Connect

    Andriollo, M.; Martinelli, G.; Morini, A.; Tortella, A.

    1998-07-01

    In EMS-MAGLEV high-speed transport systems, devices for propulsion, levitation and contactless on-board electric power transfer are combined in a single electromagnetic structure. The strong coupling among the windings affects the performance of each device and requires the utilization of numerical codes. The paper describes an overall optimization procedure, based on a suitable mathematical model of the system, which takes into account several items of the system performance. The parameters of the model are calculated by an automated sequence of FEM analyses of the configuration. Both the linear generator output characteristics and the propulsion force ripple are improved applying the procedure to a reference configuration. The results are compared with the results obtained by a sequence of partial optimizations operating separately on two different subsets of the geometric parameters.

  12. Online EM with weight-based forgetting.

    PubMed

    Celaya, Enric; Agostini, Alejandro

    2015-05-01

    In the online version of the EM algorithm introduced by Sato and Ishii ( 2000 ), a time-dependent discount factor is introduced for forgetting the effect of the old estimated values obtained with an earlier, inaccurate estimator. In their approach, forgetting is uniformly applied to the estimators of each mixture component depending exclusively on time, irrespective of the weight attributed to each unit for the observed sample. This causes an excessive forgetting in the less frequently sampled regions. To address this problem, we propose a modification of the algorithm that involves a weight-dependent forgetting, different for each mixture component, in which old observations are forgotten according to the actual weight of the new samples used to replace older values. A comparison of the time-dependent versus the weight-dependent approach shows that the latter improves the accuracy of the approximation and exhibits much greater stability. PMID:25710091

  13. Online EM with weight-based forgetting.

    PubMed

    Celaya, Enric; Agostini, Alejandro

    2015-05-01

    In the online version of the EM algorithm introduced by Sato and Ishii ( 2000 ), a time-dependent discount factor is introduced for forgetting the effect of the old estimated values obtained with an earlier, inaccurate estimator. In their approach, forgetting is uniformly applied to the estimators of each mixture component depending exclusively on time, irrespective of the weight attributed to each unit for the observed sample. This causes an excessive forgetting in the less frequently sampled regions. To address this problem, we propose a modification of the algorithm that involves a weight-dependent forgetting, different for each mixture component, in which old observations are forgotten according to the actual weight of the new samples used to replace older values. A comparison of the time-dependent versus the weight-dependent approach shows that the latter improves the accuracy of the approximation and exhibits much greater stability.

  14. Processing of Structurally Heterogeneous Cryo-EM Data in RELION.

    PubMed

    Scheres, S H W

    2016-01-01

    This chapter describes algorithmic advances in the RELION software, and how these are used in high-resolution cryo-electron microscopy (cryo-EM) structure determination. Since the presence of projections of different three-dimensional structures in the dataset probably represents the biggest challenge in cryo-EM data processing, special emphasis is placed on how to deal with structurally heterogeneous datasets. As such, this chapter aims to be of practical help to those who wish to use RELION in their cryo-EM structure determination efforts. PMID:27572726

  15. Processing of Structurally Heterogeneous Cryo-EM Data in RELION.

    PubMed

    Scheres, S H W

    2016-01-01

    This chapter describes algorithmic advances in the RELION software, and how these are used in high-resolution cryo-electron microscopy (cryo-EM) structure determination. Since the presence of projections of different three-dimensional structures in the dataset probably represents the biggest challenge in cryo-EM data processing, special emphasis is placed on how to deal with structurally heterogeneous datasets. As such, this chapter aims to be of practical help to those who wish to use RELION in their cryo-EM structure determination efforts.

  16. Application of the Chameleon Model to EM Field Momentum

    NASA Astrophysics Data System (ADS)

    Robertson, Glen A.

    2008-01-01

    The Chameleon scalar field model proposed by Khoury and Weltman presents an alternative mechanism for circumventing the constraints from local tests of gravity by mediating a fifth force for cosmological expansion, which could result in experimental signatures detectable through modest improvements of current laboratory set-ups in the vicinity of oscillating matter. In this paper, the oscillation of a dielectric by a crossed EM field is investigated in light of the Chameleon model. An EM excited Chameleon field-force equation is developed and compared to several EM experiments using the Barium Titanate based dielectric material.

  17. Environmental Education and Development Division (EM-522). Annual report, Fiscal year 1993

    SciTech Connect

    Not Available

    1993-12-31

    The Environmental Education and Development Division (EM-522) is one of three divisions within the Office of Technology Integration and Environmental Education and Development (EM-52) in Environmental Restoration and Waste Management`s (EM`s) Office of Technology Development (EM-50). The primary design criterion for EM-522 education activities is directly related to meeting EM`s goal of environmental compliance on an accelerated basis and cleanup of the 1989 inventory of inactive sites and facilities by the year 2019. Therefore, EM-522`s efforts are directed specifically toward stimulating knowledge and capabilities to achieve the goals of EM while contributing to DOE`s overall goal of increasing scientific, mathematical, and technical literacy and competency. This report discusses fiscal year 1993 activities.

  18. 7 CFR 759.6 - EM to be made available.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... Agriculture under the Plant Protection Act or the animal quarantine laws, as defined in section 2509 of the Food, Agriculture, Conservation, and Trade Act of 1990, automatically authorizes EM for production...

  19. Antibody-based affinity cryo-EM grid.

    PubMed

    Yu, Guimei; Li, Kunpeng; Jiang, Wen

    2016-05-01

    The Affinity Grid technique combines sample purification and cryo-Electron Microscopy (cryo-EM) grid preparation into a single step. Several types of affinity surfaces, including functionalized lipids monolayers, streptavidin 2D crystals, and covalently functionalized carbon surfaces have been reported. More recently, we presented a new affinity cryo-EM approach, cryo-SPIEM, which applies the traditional Solid Phase Immune Electron Microscopy (SPIEM) technique to cryo-EM. This approach significantly simplifies the preparation of affinity grids and directly works with native macromolecular complexes without need of target modifications. With wide availability of high affinity and high specificity antibodies, the antibody-based affinity grid would enable cryo-EM studies of the native samples directly from cell cultures, targets of low abundance, and unstable or short-lived intermediate states.

  20. E.M. and Hadronic Shower Simulation with FLUKA

    SciTech Connect

    Battistoni, G.; Fasso, A.; Ferrari, A.; Ranft, J.; Rubbia, A.; Sala, P.R.; /INFN, Milan /SLAC /CERN /Siegen U. /Zurich, ETH

    2005-10-03

    A description of the main features of e.m. and hadronic shower simulation models used in the FLUKA code is summarized and some recent applications are discussed. The general status of the FLUKA project is also reported.

  1. Near-atomic-resolution cryo-EM for molecular virology.

    PubMed

    Hryc, Corey F; Chen, Dong-Hua; Chiu, Wah

    2011-08-01

    Electron cryo-microscopy (cryo-EM) is a technique in structural biology that is widely used to solve the three-dimensional structures of macromolecular assemblies, close to their biological and solution conditions. Recent improvements in cryo-EM and single-particle reconstruction methodologies have led to the determination of several virus structures at near-atomic resolution (3.3 - 4.6 Å). These cryo-EM structures not only resolve the Cα backbones and side-chain densities of viral capsid proteins, but also suggest functional roles that the protein domains and some key amino acid residues play. This paper reviews the recent advances in near-atomic-resolution cryo-EM for probing the mechanisms of virus assembly and morphogenesis.

  2. NASA EM Followup of LIGO-Virgo Candidate Events

    NASA Technical Reports Server (NTRS)

    Blackburn, Lindy L.

    2011-01-01

    We present a strategy for a follow-up of LIGO-Virgo candidate events using offline survey data from several NASA high-energy photon instruments aboard RXTE, Swift, and Fermi. Time and sky-location information provided by the GW trigger allows for a targeted search for prompt and afterglow EM signals. In doing so, we expect to be sensitive to signals which are too weak to be publicly reported as astrophysical EM events.

  3. Reducing stress factors in EMS: report of a national survey.

    PubMed

    Brownstone, J E; Shatoff, D K; Duckro, P N

    1983-01-01

    The existence of stress, coping with stress, and the effects of stress in Emergency Medical Services (EMS) are highly current and hotly debated subjects. This article describes in part the results of a national survey of 25 EMS systems in 24 large metropolitan areas. The portion of the survey reported here focused on sources of stress and programs available to promote more effective coping with stres. Results are discussed generally and in terms of the services' organizational affiliations.

  4. EM Telemetry Tool for Deep Well Drilling Applications

    SciTech Connect

    Jeffrey M. Gabelmann

    2005-11-15

    This final report discusses the successful development and testing of a deep operational electromagnetic (EM) telemetry system, produced under a cooperative agreement with the United States Department of Energy's National Energy Technology Laboratory. This new electromagnetic telemetry system provides a wireless communication link between sensors deployed deep within oil and gas wells and data acquisition equipment located on the earth's surface. EM based wireless telemetry is a highly appropriate technology for oil and gas exploration in that it avoids the need for thousands of feet of wired connections. In order to achieve the project performance objectives, significant improvements over existing EM telemetry systems were made. These improvements included the development of new technologies that have improved the reliability of the communications link while extending operational depth. A key element of the new design is the incorporation of a data-fusion methodology which enhances the communication receiver's ability to extract very weak signals from large amounts of ambient environmental noise. This innovative data-fusion receiver based system adapts advanced technologies, not normally associated with low-frequency communications, and makes them work within the harsh drilling environments associated with the energy exploration market. Every element of a traditional EM telemetry system design, from power efficiency to reliability, has been addressed. The data fusion based EM telemetry system developed during this project is anticipated to provide an EM tool capability that will impact both onshore and offshore oil and gas exploration operations, for conventional and underbalanced drilling applications.

  5. A HF EM installation allowing simultaneous whole body and deep local EM hyperthermia.

    PubMed

    Mazokhin, V N; Kolmakov, D N; Lucheyov, N A; Gelvich, E A; Troshin, I I

    1999-01-01

    The structure and main features of a HF EM installation based upon a new approach for creating electromagnetic fields destined for whole body (WBH) and deep local (DLH) hyperthermia are discussed. The HF EM field, at a frequency of 13.56 MHz, is created by a coplanar capacity type applicator positioned under a distilled water filled bolus that the patient is lying on. The EM energy being released directly in the deep tissues ensures effective whole body heating to required therapeutic temperatures of up to 43.5 degrees C, whereas the skin temperature can be maintained as low as 39-40.5 degrees C. For DLH, the installation is equipped with additional applicators and a generator operating at a frequency of 40.68 MHz. High efficiency of the WBH applicator makes it possible to carry out the WBH procedure without any air-conditioning cabin. Due to this, a free access to the patient's body during the WBH treatment is provided and a simultaneous WBH/DLH or WBH/LH procedure by means of additional applicators is possible. Controllable power output in the range of 100-800 W at a frequency of 13.56 MHz and 50-350 W at a frequency of 40.68 MHz allows accurate temperature control during WBH, DLH and WBH/DLH procedures. SAR patterns created by the WBH and DLH applicators in a liquid muscle phantom and measured by means of a non-perturbing E-dipole are investigated. The scattered EM field strength measured in the vicinity of the operating installation during the WBH, DLH and WBH/DLH procedures does not exceed security standards. Examples of temperature versus time graphs in the course of WBH, DLH and WBH/DLH procedures in clinics are presented. The installation is successfully used in leading oncological institutions of Russia and Belarus, though combined WBH/DLH procedures are evidently more complicated and demand thorough planning and temperature measurements to avoid overheating. PMID:10458570

  6. A History and Informal Assessment of the <em>Slacker Astronomyem> Podcast

    NASA Astrophysics Data System (ADS)

    Price, Aaron; Gay, Pamela; Searle, Travis; Brissenden, Gina

    Slacker Astronomyem> is a weekly podcast that covers a recent astronomical news event or discovery. The show has a unique style consisting of irreverent, over-the-top humor combined with a healthy dose of hard science. According to our demographic analysis, the combination of this style and the unique podcasting distribution mechanism allows the show to reach audiences younger and busier than those reached via traditional channels. We report on the successes and challenges of the first year of the show, and provide an informal assessment of its role as a source for astronomical news and concepts for its approximately 15,500 weekly listeners.

  7. Databases and Archiving for CryoEM.

    PubMed

    Patwardhan, A; Lawson, C L

    2016-01-01

    CryoEM in structural biology is currently served by three public archives-EMDB for 3DEM reconstructions, PDB for models built from 3DEM reconstructions, and EMPIAR for the raw 2D image data used to obtain the 3DEM reconstructions. These archives play a vital role for both the structural community and the wider biological community in making the data accessible so that results may be reused, reassessed, and integrated with other structural and bioinformatics resources. The important role of the archives is underpinned by the fact that many journals mandate the deposition of data to PDB and EMDB on publication. The field is currently undergoing transformative changes where on the one hand high-resolution structures are becoming a routine occurrence while on the other hand electron tomography is enabling the study of macromolecules in the cellular context. Concomitantly the archives are evolving to best serve their stakeholder communities. In this chapter, we describe the current state of the archives, resources available for depositing, accessing, searching, visualizing and validating data, on-going community-wide initiatives and opportunities, and challenges for the future. PMID:27572735

  8. DOE EM industry programs robotics development

    SciTech Connect

    Staubly, R.; Kothari, V.

    1998-12-31

    The Office of Science and Technology (OST) manages an aggressive program for RD and D, as well as testing and evaluation for the Department of Energy`s (DOE`s) Environmental Management (EM) organization. The goal is to develop new and improved environmental restoration and waste management technologies to clean up the inventory of the DOE weapons complex faster, safer, and cheaper than is possible with currently available technologies. Robotic systems reduce worker exposure to the absolute minimum, while providing proven, cost-effective, and, for some applications, the only acceptable technique for addressing challenging problems. Development of robotic systems for remote operations occurs in three main categories: tank waste characterization and retrieval; decontamination and dismantlement; and characterization, mapping, and inspection systems. In addition, the Federal Energy Technology Center (FETC) has some other projects which fall under the heading of supporting R and D. The central objective of all FETC robotic projects is to make robotic systems more attractive by reducing costs and health risks associated with the deployment of robotic technologies in the cleanup of the nuclear weapons complex. This will be accomplished through development of robots that are cheaper, faster, safer, and more reliable, as well as more straightforward to modify/adapt and more intuitive to operate with autonomous capabilities and intelligent controls that prevent accidents and optimize task execution.

  9. Elected medical staff leaders: who needs 'em?

    PubMed

    Thompson, R E

    1994-03-01

    Authority, influence, and power are not synonyms. In working with elected medical staff leaders, a physician executive who chooses to exert authority may soon find him- or herself relatively powerless. But one who chooses to downplay authority, to influence through persuasion, and to coach leaders to lead effectively soon generates support for his or her ideas. The need to coax, cajole, explain, persuade, and "seek input" frustrates many leaders in all kinds of organizations. It would be much easier just to order people about. It's so tempting to think: "Who needs 'em? I'm the 'chief physician.' I know what needs to be done. Let's weigh anchor, take her out, and do what it takes to sail those rough, uncharted seas." If you really enjoy sailing a large ship in rough seas without a crew, go right ahead. Or if you think it makes sense to run an organization with only an executive staff and no knowledgeable middle managers, by all means let clinician leaders know that, now that you're aboard, they're just window-dressing. If you can make this approach work, well and good. Your life will be much less complicated, each day will have far fewer frustrations, and progress toward established goals will be much faster. However, given the reality of traditionally thinking physicians, it would be best to keep an up-dated resume in the locked lower left-hand drawer of your desk.

  10. Persistent topology for cryo-EM data analysis.

    PubMed

    Xia, Kelin; Wei, Guo-Wei

    2015-08-01

    In this work, we introduce persistent homology for the analysis of cryo-electron microscopy (cryo-EM) density maps. We identify the topological fingerprint or topological signature of noise, which is widespread in cryo-EM data. For low signal-to-noise ratio (SNR) volumetric data, intrinsic topological features of biomolecular structures are indistinguishable from noise. To remove noise, we employ geometric flows that are found to preserve the intrinsic topological fingerprints of cryo-EM structures and diminish the topological signature of noise. In particular, persistent homology enables us to visualize the gradual separation of the topological fingerprints of cryo-EM structures from those of noise during the denoising process, which gives rise to a practical procedure for prescribing a noise threshold to extract cryo-EM structure information from noise contaminated data after certain iterations of the geometric flow equation. To further demonstrate the utility of persistent homology for cryo-EM data analysis, we consider a microtubule intermediate structure Electron Microscopy Data (EMD 1129). Three helix models, an alpha-tubulin monomer model, an alpha-tubulin and beta-tubulin model, and an alpha-tubulin and beta-tubulin dimer model, are constructed to fit the cryo-EM data. The least square fitting leads to similarly high correlation coefficients, which indicates that structure determination via optimization is an ill-posed inverse problem. However, these models have dramatically different topological fingerprints. Especially, linkages or connectivities that discriminate one model from another, play little role in the traditional density fitting or optimization but are very sensitive and crucial to topological fingerprints. The intrinsic topological features of the microtubule data are identified after topological denoising. By a comparison of the topological fingerprints of the original data and those of three models, we found that the third model is

  11. Emergency medical service (EMS): A unique flight environment

    NASA Technical Reports Server (NTRS)

    Shively, R. Jay

    1993-01-01

    The EMS flight environment is unique in today's aviation. The pilots must respond quickly to emergency events and often fly to landing zones where they have never been before . The time from initially receiving a call to being airborne can be as little as two to three minutes. Often the EMS pilot is the only aviation professional on site, they have no operations people or other pilots to aid them in making decisons. Further, since they are often flying to accident scenes, not airports, there is often complete weather and condition information. Therefore, the initial decision that the pilot must make, accepting or declining a flight, can become very difficult. The accident rate of EMS helicopters has been relatively high over the past years. NASA-Ames research center has taken several steps in an attempt to aid EMS pilots in their decision making and situational awareness. A preflight risk assessment system (SAFE) was developed to aid pilots in their decision making, and was tested at an EMS service. The resutls of the study were promising and a second version incorporating the lessons learned is under development. A second line of research was the development of a low cost electronic chart display (ECD). This is a digital map display to help pilots maintain geographical orientation. Another thrust was undertaken in conjunction with the Aviation Safety Reporting System (ASRS). This involved publicizing the ASRS to EMS pilots and personnel, and calling each of the reporters back to gather additional information. This paper will discuss these efforts and how they may positively impact the safety of EMS operations.

  12. Persistent topology for cryo-EM data analysis.

    PubMed

    Xia, Kelin; Wei, Guo-Wei

    2015-08-01

    In this work, we introduce persistent homology for the analysis of cryo-electron microscopy (cryo-EM) density maps. We identify the topological fingerprint or topological signature of noise, which is widespread in cryo-EM data. For low signal-to-noise ratio (SNR) volumetric data, intrinsic topological features of biomolecular structures are indistinguishable from noise. To remove noise, we employ geometric flows that are found to preserve the intrinsic topological fingerprints of cryo-EM structures and diminish the topological signature of noise. In particular, persistent homology enables us to visualize the gradual separation of the topological fingerprints of cryo-EM structures from those of noise during the denoising process, which gives rise to a practical procedure for prescribing a noise threshold to extract cryo-EM structure information from noise contaminated data after certain iterations of the geometric flow equation. To further demonstrate the utility of persistent homology for cryo-EM data analysis, we consider a microtubule intermediate structure Electron Microscopy Data (EMD 1129). Three helix models, an alpha-tubulin monomer model, an alpha-tubulin and beta-tubulin model, and an alpha-tubulin and beta-tubulin dimer model, are constructed to fit the cryo-EM data. The least square fitting leads to similarly high correlation coefficients, which indicates that structure determination via optimization is an ill-posed inverse problem. However, these models have dramatically different topological fingerprints. Especially, linkages or connectivities that discriminate one model from another, play little role in the traditional density fitting or optimization but are very sensitive and crucial to topological fingerprints. The intrinsic topological features of the microtubule data are identified after topological denoising. By a comparison of the topological fingerprints of the original data and those of three models, we found that the third model is

  13. Degradation of Benzodiazepines after 120 Days of EMS Deployment

    PubMed Central

    McMullan, Jason T.; Jones, Elizabeth; Barnhart, Bruce; Denninghoff, Kurt; Spaite, Daniel; Zaleski, Erin; Silbergleit, Robert

    2014-01-01

    Introduction EMS treatment of status epilepticus improves outcomes, but the benzodiazepine best suited for EMS use is unclear, given potential high environmental temperature exposures. Objective To describe the degradation of diazepam, lorazepam, and midazolam as a function of temperature exposure and time over 120 days of storage on active EMS units. Methods Study boxes containing vials of diazepam, lorazepam, and midazolam were distributed to 4 active EMS units in each of 2 EMS systems in the southwestern United States during May–August 2011. The boxes logged temperature every minute and were stored in EMS units per local agency policy. Two vials of each drug were removed from each box at 30-day intervals and underwent high-performance liquid chromatography to determine drug concentration. Concentration was analyzed as mean (and 95%CI) percent of initial labeled concentration as a function of time and mean kinetic temperature (MKT). Results 192 samples were collected (2 samples of each drug from each of 4 units per city at 4 time-points). After 120 days, the mean relative concentration (95%CI) of diazepam was 97.0% (95.7–98.2%) and of midazolam was 99.0% (97.7–100.2%). Lorazepam experienced modest degradation by 60 days (95.6% [91.6–99.5%]) and substantial degradation at 90 days (90.3% [85.2-95.4%]) and 120 days (86.5% [80.7–92.3%]). Mean MKT was 31.6°C (95%CI 27.1–36.1). Increasing MKT was associated with greater degradation of lorazepam, but not midazolam or diazepam. Conclusions Midazolam and diazepam experienced minimal degradation throughout 120 days of EMS deployment in high-heat environments. Lorazepam experienced significant degradation over 120 days and appeared especially sensitive to higher MKT exposure. PMID:24548058

  14. Colloidal Oatmeal <em>(Avena Sativa)em> Improves Skin Barrier Through Multi-Therapy Activity.

    PubMed

    Ilnytska, Olha; Kaur, Simarna; Chon, Suhyoun; Reynertson, Kurt A; Nebus, Judith; Garay, Michelle; Mahmood, Khalid; Southall, Michael D

    2016-06-01

    Oats (Avena sativa) are a centuries-old topical treatment for a variety of skin barrier conditions, including dry skin, skin rashes, and eczema; however, few studies have investigated the actual mechanism of action for the skin barrier strengthening activity of colloidal oatmeal. Four extracts of colloidal oatmeal were prepared with various solvents and tested in vitro for skin barrier related gene expression and activity. Extracts of colloidal oatmeal were found to induce the expression of genes related to epidermal differentiation, tight junctions and lipid regulation in skin, and provide pH-buffering capacity. Colloidal oatmeal boosted the expression of multiple target genes related to skin barrier, and resulted in recovery of barrier damage in an in vitro model of atopic dermatitis. In addition, an investigator-blinded study was performed with 50 healthy female subjects who exhibited bilateral moderate to severe dry skin on their lower legs. Subjects were treated with a colloidal oatmeal skin protectant lotion. Clinically, the colloidal oatmeal lotion showed significant clinical improvements in skin dryness, moisturization, and barrier. Taken together, these results demonstrate that colloidal oatmeal can provide clinically effective benefits for dry and compromised skin by strengthening skin barrier.

    <em>J Drugs Dermatolem>. 2016;15(6):684-690. PMID:27272074

  15. Colloidal Oatmeal <em>(Avena Sativa)em> Improves Skin Barrier Through Multi-Therapy Activity.

    PubMed

    Ilnytska, Olha; Kaur, Simarna; Chon, Suhyoun; Reynertson, Kurt A; Nebus, Judith; Garay, Michelle; Mahmood, Khalid; Southall, Michael D

    2016-06-01

    Oats (Avena sativa) are a centuries-old topical treatment for a variety of skin barrier conditions, including dry skin, skin rashes, and eczema; however, few studies have investigated the actual mechanism of action for the skin barrier strengthening activity of colloidal oatmeal. Four extracts of colloidal oatmeal were prepared with various solvents and tested in vitro for skin barrier related gene expression and activity. Extracts of colloidal oatmeal were found to induce the expression of genes related to epidermal differentiation, tight junctions and lipid regulation in skin, and provide pH-buffering capacity. Colloidal oatmeal boosted the expression of multiple target genes related to skin barrier, and resulted in recovery of barrier damage in an in vitro model of atopic dermatitis. In addition, an investigator-blinded study was performed with 50 healthy female subjects who exhibited bilateral moderate to severe dry skin on their lower legs. Subjects were treated with a colloidal oatmeal skin protectant lotion. Clinically, the colloidal oatmeal lotion showed significant clinical improvements in skin dryness, moisturization, and barrier. Taken together, these results demonstrate that colloidal oatmeal can provide clinically effective benefits for dry and compromised skin by strengthening skin barrier.

    <em>J Drugs Dermatolem>. 2016;15(6):684-690.

  16. FitEM2EM--tools for low resolution study of macromolecular assembly and dynamics.

    PubMed

    Frankenstein, Ziv; Sperling, Joseph; Sperling, Ruth; Eisenstein, Miriam

    2008-01-01

    Studies of the structure and dynamics of macromolecular assemblies often involve comparison of low resolution models obtained using different techniques such as electron microscopy or atomic force microscopy. We present new computational tools for comparing (matching) and docking of low resolution structures, based on shape complementarity. The matched or docked objects are represented by three dimensional grids where the value of each grid point depends on its position with regard to the interior, surface or exterior of the object. The grids are correlated using fast Fourier transformations producing either matches of related objects or docking models depending on the details of the grid representations. The procedures incorporate thickening and smoothing of the surfaces of the objects which effectively compensates for differences in the resolution of the matched/docked objects, circumventing the need for resolution modification. The presented matching tool FitEM2EMin successfully fitted electron microscopy structures obtained at different resolutions, different conformers of the same structure and partial structures, ranking correct matches at the top in every case. The differences between the grid representations of the matched objects can be used to study conformation differences or to characterize the size and shape of substructures. The presented low-to-low docking tool FitEM2EMout ranked the expected models at the top. PMID:18974836

  17. FitEM2EM--tools for low resolution study of macromolecular assembly and dynamics.

    PubMed

    Frankenstein, Ziv; Sperling, Joseph; Sperling, Ruth; Eisenstein, Miriam

    2008-01-01

    Studies of the structure and dynamics of macromolecular assemblies often involve comparison of low resolution models obtained using different techniques such as electron microscopy or atomic force microscopy. We present new computational tools for comparing (matching) and docking of low resolution structures, based on shape complementarity. The matched or docked objects are represented by three dimensional grids where the value of each grid point depends on its position with regard to the interior, surface or exterior of the object. The grids are correlated using fast Fourier transformations producing either matches of related objects or docking models depending on the details of the grid representations. The procedures incorporate thickening and smoothing of the surfaces of the objects which effectively compensates for differences in the resolution of the matched/docked objects, circumventing the need for resolution modification. The presented matching tool FitEM2EMin successfully fitted electron microscopy structures obtained at different resolutions, different conformers of the same structure and partial structures, ranking correct matches at the top in every case. The differences between the grid representations of the matched objects can be used to study conformation differences or to characterize the size and shape of substructures. The presented low-to-low docking tool FitEM2EMout ranked the expected models at the top.

  18. Learning when to Hold'em and When to Fold'em: ERS's Budget Hold'em Game Facilitates the Budget Development Process in Memphis

    ERIC Educational Resources Information Center

    Education Resource Strategies, 2010

    2010-01-01

    If your school district is facing a budget issue, it might surprise you to learn that the solution might very well lie in a game of cards. That certainly was the case earlier this year for the city schools of Memphis, Tennessee. The game is called Budget Hold'em, and it was developed by Education Resource Strategies (ERS) of Watertown,…

  19. Refinement of Atomic Structures Against cryo-EM Maps.

    PubMed

    Murshudov, G N

    2016-01-01

    This review describes some of the methods for atomic structure refinement (fitting) against medium/high-resolution single-particle cryo-EM reconstructed maps. Some of the tools developed for macromolecular X-ray crystal structure analysis, especially those encapsulating prior chemical and structural information can be transferred directly for fitting into cryo-EM maps. However, despite the similarities, there are significant differences between data produced by these two techniques; therefore, different likelihood functions linking the data and model must be used in cryo-EM and crystallographic refinement. Although tools described in this review are mostly designed for medium/high-resolution maps, if maps have sufficiently good quality, then these tools can also be used at moderately low resolution, as shown in one example. In addition, the use of several popular crystallographic methods is strongly discouraged in cryo-EM refinement, such as 2Fo-Fc maps, solvent flattening, and feature-enhanced maps (FEMs) for visualization and model (re)building. Two problems in the cryo-EM field are overclaiming resolution and severe map oversharpening. Both of these should be avoided; if data of higher resolution than the signal are used, then overfitting of model parameters into the noise is unavoidable, and if maps are oversharpened, then at least parts of the maps might become very noisy and ultimately uninterpretable. Both of these may result in suboptimal and even misleading atomic models.

  20. International Space Station (ISS) Emergency Mask (EM) Development

    NASA Technical Reports Server (NTRS)

    Toon, Katherine P.; Hahn, Jeffrey; Fowler, Michael; Young, Kevin

    2011-01-01

    The Emergency Mask (EM) is considered a secondary response emergency Personal Protective Equipment (PPE) designed to provide respiratory protection to the International Space Station (ISS) crewmembers in response to a post-fire event or ammonia leak. The EM is planned to be delivered to ISS in 2012 to replace the current air purifying respirator (APR) onboard ISS called the Ammonia Respirator (AR). The EM is a one ]size ]fits ]all model designed to fit any size crewmember, unlike the APR on ISS, and uses either two Fire Cartridges (FCs) or two Commercial Off-the-Shelf (COTS) 3M(Trademark). Ammonia Cartridges (ACs) to provide the crew with a minimum of 8 hours of respiratory protection with appropriate cartridge swap ]out. The EM is designed for a single exposure event, for either post ]fire or ammonia, and is a passive device that cannot help crewmembers who cannot breathe on their own. The EM fs primary and only seal is around the wearer fs neck to prevent a crewmember from inhaling contaminants. During the development of the ISS Emergency Mask, several design challenges were faced that focused around manufacturing a leak free mask. The description of those challenges are broadly discussed but focuses on one key design challenge area: bonding EPDM gasket material to Gore(Registered Trademark) fabric hood.

  1. Breaking Cryo-EM Resolution Barriers to Facilitate Drug Discovery.

    PubMed

    Merk, Alan; Bartesaghi, Alberto; Banerjee, Soojay; Falconieri, Veronica; Rao, Prashant; Davis, Mindy I; Pragani, Rajan; Boxer, Matthew B; Earl, Lesley A; Milne, Jacqueline L S; Subramaniam, Sriram

    2016-06-16

    Recent advances in single-particle cryoelecton microscopy (cryo-EM) are enabling generation of numerous near-atomic resolution structures for well-ordered protein complexes with sizes ≥ ∼200 kDa. Whether cryo-EM methods are equally useful for high-resolution structural analysis of smaller, dynamic protein complexes such as those involved in cellular metabolism remains an important question. Here, we present 3.8 Å resolution cryo-EM structures of the cancer target isocitrate dehydrogenase (93 kDa) and identify the nature of conformational changes induced by binding of the allosteric small-molecule inhibitor ML309. We also report 2.8-Å- and 1.8-Å-resolution structures of lactate dehydrogenase (145 kDa) and glutamate dehydrogenase (334 kDa), respectively. With these results, two perceived barriers in single-particle cryo-EM are overcome: (1) crossing 2 Å resolution and (2) obtaining structures of proteins with sizes < 100 kDa, demonstrating that cryo-EM can be used to investigate a broad spectrum of drug-target interactions and dynamic conformational states.

  2. Refinement of Atomic Structures Against cryo-EM Maps.

    PubMed

    Murshudov, G N

    2016-01-01

    This review describes some of the methods for atomic structure refinement (fitting) against medium/high-resolution single-particle cryo-EM reconstructed maps. Some of the tools developed for macromolecular X-ray crystal structure analysis, especially those encapsulating prior chemical and structural information can be transferred directly for fitting into cryo-EM maps. However, despite the similarities, there are significant differences between data produced by these two techniques; therefore, different likelihood functions linking the data and model must be used in cryo-EM and crystallographic refinement. Although tools described in this review are mostly designed for medium/high-resolution maps, if maps have sufficiently good quality, then these tools can also be used at moderately low resolution, as shown in one example. In addition, the use of several popular crystallographic methods is strongly discouraged in cryo-EM refinement, such as 2Fo-Fc maps, solvent flattening, and feature-enhanced maps (FEMs) for visualization and model (re)building. Two problems in the cryo-EM field are overclaiming resolution and severe map oversharpening. Both of these should be avoided; if data of higher resolution than the signal are used, then overfitting of model parameters into the noise is unavoidable, and if maps are oversharpened, then at least parts of the maps might become very noisy and ultimately uninterpretable. Both of these may result in suboptimal and even misleading atomic models. PMID:27572731

  3. Analyses of Subnanometer Resolution Cryo-EM Density Maps

    PubMed Central

    Baker, Matthew L.; Baker, Mariah R.; Hryc, Corey F.; DiMaio, Frank

    2011-01-01

    Today, electron cryomicroscopy (cryo-EM) can routinely achieve subnanometer resolutions of complex macromolecular assemblies. From a density map, one can extract key structural and functional information using a variety of computational analysis tools. At subnanometer resolution, these tools make it possible to isolate individual subunits, identify secondary structures, and accurately fit atomic models. With several cryo-EM studies achieving resolutions beyond 5 Å, computational modeling and feature recognition tools have been employed to construct backbone and atomic models of the protein components directly from a density map. In this chapter, we describe several common classes of computational tools that can be used to analyze and model subnanometer resolution reconstructions from cryo-EM. A general protocol for analyzing subnanometer resolution density maps is presented along with a full description of steps used in analyzing the 4.3 Å resolution structure of Mm-cpn. PMID:20888467

  4. EMS-induced cytomictic variability in safflower (Carthamus tinctorius L.).

    PubMed

    Srivastava, P; Kumar, G

    2011-01-01

    Seeds of safflower (Carthamus tinctorius L.) were subjected to three treatment durations (3h, 5h and 7h) of 0.5 % Ethyl Methane Sulphonate (EMS). Microsporogenesis was carried out in the control as well as in the treated materials. EMS treated plants showed interesting feature of partial inter-meiocyte chromatin migration through channel formation, beak formation or direct cell fusion. Another interesting feature noticed during the study was the fusion among tetrads due to wall dissolution. The phenomenon of cytomixis was recorded at nearly all the stages of microsporogenesis connecting from a few to several meiocytes. Other abnormalities such as laggards, precocious movement, bridge and non-disjunction of chromosomes were also recorded but in very low frequencies. The phenomenon of cytomixis increased along with the increase in treatment duration of EMS. Cells with these types of cytomictic disturbances may probably result in uneven formation of gametes or zygote, heterogenous sized pollen grains or even loss of fertility in future.

  5. Virus particle dynamics derived from CryoEM studies.

    PubMed

    Doerschuk, Peter C; Gong, Yunye; Xu, Nan; Domitrovic, Tatiana; Johnson, John E

    2016-06-01

    The direct electron detector has revolutionized electron cryo-microscopy (CryoEM). Icosahedral virus structures are routinely produced at 4Å resolution or better and the approach has largely displaced virus crystallography, as it requires less material, less purity and often produces a structure more rapidly. Largely ignored in this new era of CryoEM is the dynamic information in the data sets that was not available in X-ray structures. Here we review an approach that captures the dynamic character of viruses displayed in the CryoEM ensemble of particles at the moment of freezing. We illustrate the approach with a simple model, briefly describe the details and provide a practical application to virus particle maturation. PMID:27085980

  6. Recent technical advancements enabled atomic resolution CryoEM

    NASA Astrophysics Data System (ADS)

    Xueming, Li

    2016-01-01

    With recent breakthroughs in camera and image processing technologies single-particle electron cryo-microscopy (CryoEM) has suddenly gained the attention of structural biologists as a powerful tool able to solve the atomic structures of biological complexes and assemblies. Compared with x-ray crystallography, CryoEM can be applied to partially flexible structures in solution and without the necessity of crystallization, which is especially important for large complexes and assemblies. This review briefly explains several key bottlenecks for atomic resolution CryoEM, and describes the corresponding solutions for these bottlenecks based on the recent technical advancements. The review also aims to provide an overview about the technical differences between its applications in biology and those in material science. Project supported by Tsinghua-Peking Joint Center for Life Sciences, China.

  7. The effective molarity (EM)--a computational approach.

    PubMed

    Karaman, Rafik

    2010-08-01

    The effective molarity (EM) for 12 intramolecular S(N)2 processes involving the formation of substituted aziridines and substituted epoxides were computed using ab initio and DFT calculation methods. Strong correlation was found between the calculated effective molarity and the experimentally determined values. This result could open a door for obtaining EM values for intramolecular processes that are difficult to be experimentally provided. Furthermore, the calculation results reveal that the driving forces for ring-closing reactions in the two different systems are proximity orientation of the nucleophile to the electrophile and the ground strain energies of the products and the reactants.

  8. Geospatial Analysis of Pediatric EMS Run Density and Endotracheal Intubation

    PubMed Central

    Hansen, Matthew; Loker, William; Warden, Craig

    2016-01-01

    Introduction The association between geographic factors, including transport distance, and pediatric emergency medical services (EMS) run clustering on out-of-hospital pediatric endotracheal intubation is unclear. The objective of this study was to determine if endotracheal intubation procedures are more likely to occur at greater distances from the hospital and near clusters of pediatric calls. Methods This was a retrospective observational study including all EMS runs for patients less than 18 years of age from 2008 to 2014 in a geographically large and diverse Oregon county that includes densely populated urban areas near Portland and remote rural areas. We geocoded scene addresses using the automated address locator created in the cloud-based mapping platform ArcGIS, supplemented with manual address geocoding for remaining cases. We then use the Getis-Ord Gi spatial statistic feature in ArcGIS to map statistically significant spatial clusters (hot spots) of pediatric EMS runs throughout the county. We then superimposed all intubation procedures performed during the study period on maps of pediatric EMS-run hot spots, pediatric population density, fire stations, and hospitals. We also performed multivariable logistic regression to determine if distance traveled to the hospital was associated with intubation after controlling for several confounding variables. Results We identified a total of 7,797 pediatric EMS runs during the study period and 38 endotracheal intubations. In univariate analysis we found that patients who were intubated were similar to those who were not in gender and whether or not they were transported to a children’s hospital. Intubated patients tended to be transported shorter distances and were older than non-intubated patients. Increased distance from the hospital was associated with reduced odds of intubation after controlling for age, sex, scene location, and trauma system entry status in a multivariate logistic regression. The

  9. Speaker verification using combined acoustic and EM sensor signal processing

    SciTech Connect

    Ng, L C; Gable, T J; Holzrichter, J F

    2000-11-10

    Low Power EM radar-like sensors have made it possible to measure properties of the human speech production system in real-time, without acoustic interference. This greatly enhances the quality and quantity of information for many speech related applications. See Holzrichter, Burnett, Ng, and Lea, J. Acoustic. SOC. Am . 103 ( 1) 622 (1998). By combining the Glottal-EM-Sensor (GEMS) with the Acoustic-signals, we've demonstrated an almost 10 fold reduction in error rates from a speaker verification system experiment under a moderate noisy environment (-10dB).

  10. Low Bandwidth Vocoding using EM Sensor and Acoustic Signal Processing

    SciTech Connect

    Ng, L C; Holzrichter, J F; Larson, P E

    2001-10-25

    Low-power EM radar-like sensors have made it possible to measure properties of the human speech production system in real-time, without acoustic interference [1]. By combining these data with the corresponding acoustic signal, we've demonstrated an almost 10-fold bandwidth reduction in speech compression, compared to a standard 2.4 kbps LPC10 protocol used in the STU-III (Secure Terminal Unit, third generation) telephone. This paper describes a potential EM sensor/acoustic based vocoder implementation.

  11. Waste fuel, EMS may save plant $1M yearly

    SciTech Connect

    Barber, J.

    1982-05-24

    A mixture of paper trash and coal ash fueling an Erie, Pa. General Electric plant and a Network 90 microprocessor-based energy-management system (EMS) to optimize boiler efficiency will cost about $3 million and have a three-to-four-year payback. Over half the savings will come from the avoided costs of burning plant-generated trash. The EMS system will monitor fuel requirements in the boiler and compensate for changes in steam demand. It will also monitor plant electrical needs and control the steam diverted for cogeneration. (DCK)

  12. A emissão em 8mm e as bandas de Merrill-Sanford em estrelas carbonadas

    NASA Astrophysics Data System (ADS)

    de Mello, A. B.; Lorenz-Martins, S.

    2003-08-01

    Estrelas carbonadas possuem bandas moleculares em absorção no visível e, no infravermelho (IR) as principais características espectrais se devem a emissão de grãos. Recentemente foi detectada a presença de bandas de SiC2 (Merrill-Sanford, MS) em emissão sendo atribuída à presença de um disco rico em poeira. Neste trabalho analisamos uma amostra de 14 estrelas carbonadas, observadas no telescópio de 1.52 m do ESO em 4 regiões espectrais diferentes, a fim de detectar as bandas de MS em emissão. Nossa amostra é composta de estrelas que apresentam além da emissão em 11.3 mm, outra em 8 mm. Esta última emissão, não usual nestes objetos, tem sido atribuída ou a moléculas de C2H2, ou a um composto sólido ainda indefinido. A detecção de emissões de MS e aquelas no IR, simultaneamente, revelaria um cenário mais complexo que o habitualmente esperado para os ventos destes objetos. No entanto como primeiro resultado, verificamos que as bandas de Merrill-Sanford encontram-se em absorção, não revelando nenhuma conexão com a emissão a 8 mm. Assim, temos duas hipóteses: (a) a emissão a 8 mm se deve à molécula C2H2 ou (b) essa emissão é resultado da emissão térmica de grãos. Testamos a segunda hipótese modelando a amostra com grãos não-homogêneos de SiC e quartzo, o qual emite em aproximadamente 8mm. Este grão seria produzido em uma fase evolutiva anterior a das carbonadas (estrelas S) e por terem uma estrutura cristalina são destruídos apenas na presença de campos de radiação ultravioleta muito intensos. Os modelos para os envoltórios utilizam o método de Monte Carlo para descrever o problema do transporte da radiação. As conclusões deste trabalho são: (1) as bandas de Merrill-Sanford se encontram em absorção, sugerindo um cenário usual para os ventos das estrelas da amostra; (2) neste cenário, a emissão em 8 mm seria resultado de grãos de quartzo com mantos de SiC, indicando que o quartzo poderia sobreviver a fase

  13. A modified EM algorithm for estimation in generalized mixed models.

    PubMed

    Steele, B M

    1996-12-01

    Application of the EM algorithm for estimation in the generalized mixed model has been largely unsuccessful because the E-step cannot be determined in most instances. The E-step computes the conditional expectation of the complete data log-likelihood and when the random effect distribution is normal, this expectation remains an intractable integral. The problem can be approached by numerical or analytic approximations; however, the computational burden imposed by numerical integration methods and the absence of an accurate analytic approximation have limited the use of the EM algorithm. In this paper, Laplace's method is adapted for analytic approximation within the E-step. The proposed algorithm is computationally straightforward and retains much of the conceptual simplicity of the conventional EM algorithm, although the usual convergence properties are not guaranteed. The proposed algorithm accommodates multiple random factors and random effect distributions besides the normal, e.g., the log-gamma distribution. Parameter estimates obtained for several data sets and through simulation show that this modified EM algorithm compares favorably with other generalized mixed model methods.

  14. Functionalized anatomical models for EM-neuron Interaction modeling

    NASA Astrophysics Data System (ADS)

    Neufeld, Esra; Cassará, Antonino Mario; Montanaro, Hazael; Kuster, Niels; Kainz, Wolfgang

    2016-06-01

    The understanding of interactions between electromagnetic (EM) fields and nerves are crucial in contexts ranging from therapeutic neurostimulation to low frequency EM exposure safety. To properly consider the impact of in vivo induced field inhomogeneity on non-linear neuronal dynamics, coupled EM-neuronal dynamics modeling is required. For that purpose, novel functionalized computable human phantoms have been developed. Their implementation and the systematic verification of the integrated anisotropic quasi-static EM solver and neuronal dynamics modeling functionality, based on the method of manufactured solutions and numerical reference data, is described. Electric and magnetic stimulation of the ulnar and sciatic nerve were modeled to help understanding a range of controversial issues related to the magnitude and optimal determination of strength-duration (SD) time constants. The results indicate the importance of considering the stimulation-specific inhomogeneous field distributions (especially at tissue interfaces), realistic models of non-linear neuronal dynamics, very short pulses, and suitable SD extrapolation models. These results and the functionalized computable phantom will influence and support the development of safe and effective neuroprosthetic devices and novel electroceuticals. Furthermore they will assist the evaluation of existing low frequency exposure standards for the entire population under all exposure conditions.

  15. Texas Hold 'em Online Poker: A Further Examination

    ERIC Educational Resources Information Center

    Hopley, Anthony A. B.; Dempsey, Kevin; Nicki, Richard

    2012-01-01

    Playing Texas Hold 'em Online Poker (THOP) is on the rise. However, there is relatively little research examining factors that contribute to problem gambling in poker players. The aim of this study was to extend the research findings of Hopley and Nicki (2010). The negative mood states of depression, anxiety and stress were found to be linked to…

  16. Signs and Guides: Wayfinding Alternatives for the EMS Library.

    ERIC Educational Resources Information Center

    Johnson, Johanna H.

    Concerned with increasing the accessibility of the collection of the Engineering/Math Sciences (EMS) Library at the University of California at Los Angeles through the use of self guidance systems, this practical study focused on the problem context, general library guides, and library signage in reviewing the literature, and conducted a survey of…

  17. A shape constrained MAP-EM algorithm for colorectal segmentation

    NASA Astrophysics Data System (ADS)

    Wang, Huafeng; Li, Lihong; Song, Bowen; Han, Fangfang; Liang, Zhengrong

    2013-02-01

    The task of effectively segmenting colon areas in CT images is an important area of interest in medical imaging field. The ability to distinguish the colon wall in an image from the background is a critical step in several approaches for achieving larger goals in automated computer-aided diagnosis (CAD). The related task of polyp detection, the ability to determine which objects or classes of polyps are present in a scene, also relies on colon wall segmentation. When modeling each tissue type as a conditionally independent Gaussian distribution, the tissue mixture fractions in each voxel via the modeled unobservable random processes of the underlying tissue types can be estimated by maximum a posteriori expectation-maximization (MAP-EM) algorithm in an iterative manner. This paper presents, based on the assumption that the partial volume effect (PVE) could be fully described by a tissue mixture model, a theoretical solution to the MAP-EM segmentation algorithm. However, the MAP-EM algorithm may miss some small regions which also belong to the colon wall. Combining with the shape constrained model, we present an improved algorithm which is able to merge similar regions and reserve fine structures. Experiment results show that the new approach can refine the jagged-like boundaries and achieve better results than merely exploited our previously presented MAP-EM algorithm.

  18. Airborne EM for geothermal and hydrogeological mapping

    NASA Astrophysics Data System (ADS)

    Menghini, A.; Manzella, A.; Viezzoli, A.; Montanari, D.; Maggi, S.

    2012-12-01

    Within the "VIGOR" project, aimed at assessing the geothermal potential of four regions in southern Italy, Airborne EM data have been acquired, modeled and interpreted. The system deployed was SkyTEM, a time-domain helicopter electromagnetic system designed for hydrogeophysical, environmental and mineral investigations. The AEM data provide, after data acquisition, analysis, processing, and modeling, a distribution volume of electrical resistivity, spanning an investigation depth from ground surface of few hundred meters, depending on resistivity condition. Resistivity is an important physical parameter for geothermal investigation, since it proved to be very effective in mapping anomalies due to hydrothermal fluid circulation, which usually has high salt content and produces clayey alteration minerals. Since the project required, among other issues, to define geothermal resources at shallow level, it was decided to perform a test with an airborne electromagnetic geophysical survey, to verify the advantages offered by the system in covering large areas in a short time. The geophysical survey was carried out in Sicily, Italy, in late 2011, over two test sites named "Termini" and "Western Sicily". The two areas were chosen on different basis. "Termini" area is covered by extensive geological surveys, and was going to be investigated also by means of electrical tomography in its northern part. Since geological condition of Sicily, even at shallow depth, is very complex, this area provided a good place for defining the resistivity values of the main geological units outcropping in the region. "Termini" survey has been also an occasion to define relations between resistivity distribution, lithological units and thermal conductivity. The "Western Sicily" area cover the main thermal manifestations of western Sicily, and the research target was to establish whether they are characterized by common hydrogeological or tectonic features that could be mapped by resistivity

  19. <em>An Amphibious Magnetotelluric Investigation of the Cascadian Seismogenic and ETS zones.em>

    NASA Astrophysics Data System (ADS)

    Parris, B. A.; Livelybrooks, D.; Bedrosian, P.; Egbert, G. D.; Key, K.; Schultz, A.; Cook, A.; Kant, M.; Wogan, N.; Zeryck, A.

    2015-12-01

    The amphibious Magnetotelluric Observations of Cascadia using a Huge Array (MOCHA) experiment seeks to address unresolved questions about the seismogenic locked zone and down-dip transition zone where episodic tremor and slip (ETS) originates. The presence of free fluids is thought to be one of the primary controls on ETS behavior within the Cascadia margin. Since the bulk electrical conductivity in the crust and mantle can be greatly increased by fluids, magnetotelluric(MT) observations can offer unique insights on the fluid distribution and its relation to observed ETS behavior. Here we present preliminary results from the 146 MT stations collected for the MOCHA project. MOCHA is unique in that it is the first amphibious array of MT stations occupied to provide for 3-D interpretation of conductivity structure of a subduction zone. The MOCHA data set comprises 75 onshore stations and 71 offshore stations, accumulated over a two-year period, and located on an approximate 25km grid, spanning from the trench to the Eastern Willamette Valley, and from central Oregon into middle Washington. We present the results of a series of east-west (cross-strike) oriented, two-dimensional inversions created using the MARE2DEM software that provide an initial picture of the conductivity structure of the locked and ETS zones and its along strike variations. Our models can be used to identify correlations between ETS occurrence rates and inferred fluid concentrations. Our modeling explores the impact of various parameterizations on 2-D inversion results, including inclusion of a smoothness penalty reduction along the inferred slab interface. This series of 2-D inversions can then be used collectively to help make and guide an a priori 3-D inversion. In addition we will present a preliminary 3-D inversion of the onshore stations created using the ModEM software. We are currently working on modifying ModEM to support inversion of offshore data. The more computationally intensive 3-D

  20. The B and Be States of the Star EM Cepheus

    NASA Astrophysics Data System (ADS)

    Kjurkchieva, Diana; Marchev, Dragomir; Sigut, T. A. A.; Dimitrov, Dinko

    2016-09-01

    We present 11 yr of high-resolution, spectroscopic observations for the star EM Cep. EM Cep switches between B and Be star states, as revealed by the level of Hα emission, but spends most of its time in the B star state. EM Cep has been considered to be an eclipsing, near-contact binary of nearly equal-mass B stars in order to reproduce regular photometric variations; however, this model is problematic due to the lack of any observed Doppler shift in the spectrum. Our observations confirm that there are no apparent Doppler shifts in the wide spectral lines Hα and He i λ6678 in either the B or Be star states. The profiles of He i λ6678 typically exhibited a filled-in absorption core, but we detected weak emission in this line during the highest Be state. Given the lack of observed Doppler shifts, we model EM Cep as an isolated Be star with a variable circumstellar disk. We can reproduce the observed Hα emission profiles over the 11 yr period reasonably well with disk masses on the order of 3{--}10× {10}-11 {M}* in the Be state with the circumstellar disk seen at an inclination of 78° to the line of sight. From a disk ejection episode in 2014, we estimate a mass-loss rate of ≈ 3× {10}-9 {M}⊙ {{yr}}-1. The derived disk density parameters are typical of those found for the classical Be stars. We therefore suggest that the EM Cep is a classical Be star and that its photometric variations are the result of β Cep or nonradial pulsations.

  1. Signals Attenuation and Application of EM-MWD in China

    NASA Astrophysics Data System (ADS)

    Wang, R.; Liu, H.; Yang, Q.; Li, J.; Wu, D.

    2012-12-01

    In the course of drilling utilizes directional well and horizontal well, it is indispensable to master timely and exact geological guide information, and offering services for such complicated drilling is electromagnetic Measurement While Drilling (EM-MWD) which can measure guide data of drilling tool in the hole and formation data while drilling. This knowledge allows the directional driller to make appropriate mechanical corrections in drill string orientation that will allow the advancing drill bit to hit an intended subsurface target area. Based on electromagnetic field theory, the paper has studied the propagation particularity and attenuation regularity that the signals of electromagnetic wave for EM-MWD transmit in stratum. The paper also gives a brief introduction of the containing, the work principle and the main technology parameter of EM-MWD. To check up the performance of EM-MWD, the field test of 5 wells were carried out in Shengli Oilfield and Liaohe Oilfield. Numerical simulation results indicate the signal attenuation will be added with the decrease of the stratum resistibility. In the frequency range from 1 to 10 Hz the stratum absorptivity is tiny and does not add noticeably with the increase of the electromagnetic wave frequency and the decrease of the stratum resistibility. In the frequency range from 1 to 10000 Hz the stratum absorptivity does not increase noticeably with the decrease of the dielectric constant of the stratum. Field test demonstrate that EM-MWD receives successfully signal emitted from depth underground 1600 meters in Shengli oilfield and 2400 meters in Liaohe oilfield. Our results indicate that numerical simulation methods are comparable to field test.

  2. The B and Be States of the Star EM Cepheus

    NASA Astrophysics Data System (ADS)

    Kjurkchieva, Diana; Marchev, Dragomir; Sigut, T. A. A.; Dimitrov, Dinko

    2016-09-01

    We present 11 yr of high-resolution, spectroscopic observations for the star EM Cep. EM Cep switches between B and Be star states, as revealed by the level of Hα emission, but spends most of its time in the B star state. EM Cep has been considered to be an eclipsing, near-contact binary of nearly equal-mass B stars in order to reproduce regular photometric variations; however, this model is problematic due to the lack of any observed Doppler shift in the spectrum. Our observations confirm that there are no apparent Doppler shifts in the wide spectral lines Hα and He i λ6678 in either the B or Be star states. The profiles of He i λ6678 typically exhibited a filled-in absorption core, but we detected weak emission in this line during the highest Be state. Given the lack of observed Doppler shifts, we model EM Cep as an isolated Be star with a variable circumstellar disk. We can reproduce the observed Hα emission profiles over the 11 yr period reasonably well with disk masses on the order of 3{--}10× {10}-11 {M}* in the Be state with the circumstellar disk seen at an inclination of 78° to the line of sight. From a disk ejection episode in 2014, we estimate a mass-loss rate of ≈ 3× {10}-9 {M}ȯ {{yr}}-1. The derived disk density parameters are typical of those found for the classical Be stars. We therefore suggest that the EM Cep is a classical Be star and that its photometric variations are the result of β Cep or nonradial pulsations.

  3. Composting of rice straw with effective microorganisms (EM) and its influence on compost quality

    PubMed Central

    2013-01-01

    This study aims to assess the effect of EM application on the composting process of rice straw with goat manure and green waste and to evaluate the quality of both compost treatments. There are two treatment piles in this study, in which one pile was applied with EM and another pile without EM. Each treatment was replicated three times with 90 days of composting duration. The parameters for the temperature, pH, TOC and C/N ratio, show that decomposition of organic matter occurs during the 90-day period. The t-test conducted shows that there is a significant difference between compost with EM and compost without EM. The application of EM in compost increases the macro and micronutrient content. The following parameters support this conclusion: compost applied with EM has more N, P and K content (P < 0.05) compared to compost without EM. Although the Fe in compost with EM is much higher (P < 0.05) than in the compost without EM, for Zn and Cu, there is no significant difference between treatments. This study suggests that the application of EM is suitable to increase the mineralization in the composting process. The final resultant compost indicated that it was in the range of the matured level and can be used without any restriction. PMID:23390930

  4. Application of Electromagnetic (EM) Separation Technology to Metal Refining Processes: A Review

    NASA Astrophysics Data System (ADS)

    Zhang, Lifeng; Wang, Shengqian; Dong, Anping; Gao, Jianwei; Damoah, Lucas Nana Wiredu

    2014-12-01

    Application of electromagnetic (EM) force to metal processing has been considered as an emerging technology for the production of clean metals and other advanced materials. In the current paper, the principle of EM separation was introduced and several schemes of imposing EM field, such as DC electric field with a crossed steady magnetic field, AC electric field, AC magnetic field, and traveling magnetic field were reviewed. The force around a single particle or multi-particles and their trajectories in the conductive liquid under EM field were discussed. Applications of EM technique to the purification of different liquid metals such as aluminum, zinc, magnesium, silicon, copper, and steel were summarized. Effects of EM processing parameters, such as the frequency of imposed field, imposed magnetic flux density, processing time, particle size, and the EM unit size on the EM purification efficiency were discussed. Experimental and theoretical investigations have showed that the separation efficiency of inclusions from the molten aluminum using EM purification could as high as over 90 pct. Meanwhile, the EM purification was also applied to separate intermetallic compounds from metal melt, such as α-AlFeMnSi-phase from the molten aluminum. And then the potential industrial application of EM technique was proposed.

  5. Effects of caffeine or EDTA post-treatment on EMS mutagenesis in soybean.

    PubMed

    Zhu, B; Gu, A; Deng, X; Geng, Y; Lu, Z

    1995-04-01

    Seeds of soybean cultivar LD4 were mutagenically treated with EMS (0.3, 0.5, 0.6, 0.9, 1.5 and 1.8%) for 3 h only or plus caffeine (50 mM) or EDTA (1 mM) post-treatment for 5 h. The experimental results indicated that: (1) of the different concentrations of EMS treatment, the M2 mutation frequency induced with 0.6% EMS was the highest (9.7%). When the EMS concentration was over 0.9%, the mutation frequency decreased rapidly. (2) Of the EMS treatments plus caffeine or EDTA post-treatment, the mutagenic effect of 0.6% EMS was the best for inducing morphological variations. Caffeine post-treatment decreased notably the mutation frequency of EMS treatment; when concentrations of EMS were very high (1.5% and 1.8%), mutation frequencies of EDTA post-treatment were still 5.0% and 4.88%, but no mutants were found in EMS treatment or plus caffeine post-treatment. (3) In the M2 mutation spectrum, 11 kinds of mutant types were observed in EMS treatment or plus caffeine or EDTA post-treatment. Relative frequencies of some mutant types (growth period, plant height, grain size, leaf shape and sterility, etc.) were similar among the three treatments, but EDTA post-treatment could change the relative frequencies of yield characteristics (number of pods and grains, grain weight/plant) induced by EMS treatment only.

  6. Development of the Emergency Medical Services Role Identity Scale (EMS-RIS).

    PubMed

    Donnelly, Elizabeth A; Siebert, Darcy; Siebert, Carl

    2015-01-01

    This article describes the development and validation of the theoretically grounded Emergency Medical Services Role Identity Scale (EMS-RIS), which measures four domains of EMS role identity. The EMS-RIS was developed using a mixed methods approach. Key informants informed item development and the scale was validated using a representative probability sample of EMS personnel. Factor analyses revealed a conceptually consistent, four-factor solution with sound psychometric properties as well as evidence of convergent and discriminant validities. Social workers work with EMS professionals in crisis settings and as their counselors when they are distressed. The EMS-RIS provides useful information for the assessment of and intervention with distressed EMS professionals, as well as how role identity may influence occupational stress. PMID:25760489

  7. Similarity-regulation of OS-EM for accelerated SPECT reconstruction

    NASA Astrophysics Data System (ADS)

    Vaissier, P. E. B.; Beekman, F. J.; Goorden, M. C.

    2016-06-01

    Ordered subsets expectation maximization (OS-EM) is widely used to accelerate image reconstruction in single photon emission computed tomography (SPECT). Speedup of OS-EM over maximum likelihood expectation maximization (ML-EM) is close to the number of subsets used. Although a high number of subsets can shorten reconstruction times significantly, it can also cause severe image artifacts such as improper erasure of reconstructed activity if projections contain few counts. We recently showed that such artifacts can be prevented by using a count-regulated OS-EM (CR-OS-EM) algorithm which automatically adapts the number of subsets for each voxel based on the estimated number of counts that the voxel contributed to the projections. While CR-OS-EM reached high speed-up over ML-EM in high-activity regions of images, speed in low-activity regions could still be very slow. In this work we propose similarity-regulated OS-EM (SR-OS-EM) as a much faster alternative to CR-OS-EM. SR-OS-EM also automatically and locally adapts the number of subsets, but it uses a different criterion for subset regulation: the number of subsets that is used for updating an individual voxel depends on how similar the reconstruction algorithm would update the estimated activity in that voxel with different subsets. Reconstructions of an image quality phantom and in vivo scans show that SR-OS-EM retains all of the favorable properties of CR-OS-EM, while reconstruction speed can be up to an order of magnitude higher in low-activity regions. Moreover our results suggest that SR-OS-EM can be operated with identical reconstruction parameters (including the number of iterations) for a wide range of count levels, which can be an additional advantage from a user perspective since users would only have to post-filter an image to present it at an appropriate noise level.

  8. Em polypeptide and its messenger RNA levels are modulated by abscisic acid during embryogenesis in wheat.

    PubMed

    Williamson, J D; Quatrano, R S; Cuming, A C

    1985-10-15

    The effect of abscisic acid (ABA) on the expression of the 'early-methionine-labeled' (Em) polypeptide was examined in cultured, immature wheat (Triticum aestivum, L.) embryos and in developing embryos in planta. A complementary DNA (cDNA) library was constructed from poly(A)-rich RNA from immature embryos cultured in the presence of ABA. ABA-enhanced sequences were first identified by differential colony-blot hybridization, and then verified using RNA slot-blot analysis. Dot-blot hybridization showed that one clone, p1015, was homologous to the previously isolated Em cDNA, pWG432. Electrophoretic analysis of the hybrid-select translation product of p1015 confirmed its identity as an Em sequence. Comparison of the p1015 cDNA insert size and the Em message size, from northern blot analysis, showed that p1015 contained about 87% of the Em sequence. RNA slot-blot analysis and protein electrophoresis showed that Em message, but not Em protein, accumulated at a low, basal level in immature embryos in the absence of ABA. Neither Em message nor Em protein was seen in three-day germinated seedlings. Steady-state levels of Em message and protein increased in immature embryos in the presence of ABA, both in culture and in planta. Regulation appeared to be primarily at the level of transcription or specific message stability. Regulation may also involve specific protein stability, since synthesis of Em protein continued in immature embryos in the absence of ABA, but Em protein did not accumulate in detectable amounts. We conclude that ABA specifically modulates Em message and protein levels in immature embryos, but is probably not responsible for the embryogenic specificity of Em expression.

  9. Magen David Adom--the EMS in Israel.

    PubMed

    Ellis, Daniel Y; Sorene, Eliot

    2008-01-01

    Israel is a small country with a population of around 7 million. The sole EMS provider for Israel is Magen David Adom (MDA) (translated as 'Red Shield of David'). MDA also carries out the functions of a National Society (similar to the Red Cross) and provides all the blood and blood product services for the country. Nationwide, the organisation responds to over 1000 emergency calls a day and uses doctors, paramedics, emergency medical technicians and volunteers. Local geopolitics has meant that MDA has to be prepared for anything from everyday emergency calls to suicide bombings and regional wars. MDA also prides itself in being able to rapidly assemble and dispatch mobile aid teams to scenes of international disasters. Such a broad range of activities is unusual for a single EMS organisation. PMID:17767990

  10. Magen David Adom--the EMS in Israel.

    PubMed

    Ellis, Daniel Y; Sorene, Eliot

    2008-01-01

    Israel is a small country with a population of around 7 million. The sole EMS provider for Israel is Magen David Adom (MDA) (translated as 'Red Shield of David'). MDA also carries out the functions of a National Society (similar to the Red Cross) and provides all the blood and blood product services for the country. Nationwide, the organisation responds to over 1000 emergency calls a day and uses doctors, paramedics, emergency medical technicians and volunteers. Local geopolitics has meant that MDA has to be prepared for anything from everyday emergency calls to suicide bombings and regional wars. MDA also prides itself in being able to rapidly assemble and dispatch mobile aid teams to scenes of international disasters. Such a broad range of activities is unusual for a single EMS organisation.

  11. Advanced communication infrastructure for pre-hospital EMS care.

    PubMed

    Orthner, Helmuth; Mazza, Giovanni; Mazza, Giovanni Giorgio; Shenvi, Rohit; Battles, Marcie

    2008-11-06

    The traditional communication infrastructure of the pre-hospital Emergency Medical System (EMS) is limited to voice communication using radio or cell phone technologies. With the emergence of 3rd Generation wireless networks (3G) and enhanced mobile devices capable of data communication (e.g., mobile tablets, PDAs with cell phones, or cell phones with PDA capabilities), the voice communication can be enhanced with interactive data messaging and perhaps even with interactive video communication. However, video requires substantially more bandwidth which 4th Generation (4G) systems are promising. However, their availability is limited. We present an infrastructure that allows dynamic selection of the best data transport mode in the pre-hospital EMS environment.

  12. Superconducting Electromagnetic Suspension (EMS) system for Grumman Maglev concept

    NASA Technical Reports Server (NTRS)

    Kalsi, Swarn S.

    1994-01-01

    The Grumman developed Electromagnetic Suspension (EMS) Maglev system has the following key characteristics: a large operating airgap--40 mm; levitation at all speeds; both high speed and low speed applications; no deleterious effects on SC coils at low vehicle speeds; low magnetic field at the SC coil--less than 0.35 T; no need to use non-magnetic/non-metallic rebar in the guideway structure; low magnetic field in passenger cabin--approximately 1 G; low forces on the SC coil; employs state-of-the-art NbTi wire; no need for an active magnet quench protection system; and lower weight than a magnet system with copper coils. The EMS Maglev described in this paper does not require development of any new technologies. The system could be built with the existing SC magnet technology.

  13. Range Condition and ML-EM Checkerboard Artifacts

    PubMed Central

    You, Jiangsheng; Wang, Jing; Liang, Zhengrong

    2007-01-01

    The expectation maximization (EM) algorithm for the maximum likelihood (ML) image reconstruction criterion generates severe checkerboard artifacts in the presence of noise. A classical remedy is to impose an a priori constraint for a penalized ML or maximum a posteriori probability solution. The penalty reduces the checkerboard artifacts and also introduces uncertainty because a priori information is usually unknown in clinic. Recent theoretical investigation reveals that the noise can be divided into two components: one is called null-space noise and the other is range-space noise. The null-space noise can be numerically estimated using filtered backprojection (FBP) algorithm. By the FBP algorithm, the null-space noise annihilates in the reconstruction while the range-space noise propagates into the reconstructed image. The aim of this work is to investigate the relation between the null-space noise and the checkerboard artifacts in the ML-EM reconstruction from noisy projection data. Our study suggests that removing the null-space noise from the projection data could improve the signal-to-noise ratio of the projection data and, therefore, reduce the checkerboard artifacts in the ML-EM reconstructed images. This study reveals an in-depth understanding of the different noise propagations in analytical and iterative image reconstructions, which may be useful to single photon emission computed tomography, where the noise has been a major factor for image degradation. The reduction of the ML-EM checkerboard artifacts by removing the null-space noise avoids the uncertainty of using a priori penalty. PMID:18449363

  14. Generalized single-particle cryo-EM--a historical perspective.

    PubMed

    Frank, Joachim

    2016-02-01

    This is a brief account of the earlier history of single-particle cryo-EM of biological molecules lacking internal symmetry, which goes back to the mid-seventies. The emphasis of this review is on the mathematical concepts and computational approaches. It is written as the field experiences a turning point in the wake of the introduction of digital cameras capable of single electron counting, and near-atomic resolution can be reached even for smaller molecules. PMID:26566976

  15. A Bayesian View on Cryo-EM Structure Determination

    PubMed Central

    Scheres, Sjors H.W.

    2012-01-01

    Three-dimensional (3D) structure determination by single-particle analysis of cryo-electron microscopy (cryo-EM) images requires many parameters to be determined from extremely noisy data. This makes the method prone to overfitting, that is, when structures describe noise rather than signal, in particular near their resolution limit where noise levels are highest. Cryo-EM structures are typically filtered using ad hoc procedures to prevent overfitting, but the tuning of arbitrary parameters may lead to subjectivity in the results. I describe a Bayesian interpretation of cryo-EM structure determination, where smoothness in the reconstructed density is imposed through a Gaussian prior in the Fourier domain. The statistical framework dictates how data and prior knowledge should be combined, so that the optimal 3D linear filter is obtained without the need for arbitrariness and objective resolution estimates may be obtained. Application to experimental data indicates that the statistical approach yields more reliable structures than existing methods and is capable of detecting smaller classes in data sets that contain multiple different structures. PMID:22100448

  16. EMS-induced cytomictic variability in safflower (Carthamus tinctorius L.).

    PubMed

    Srivastava, P; Kumar, G

    2011-01-01

    Seeds of safflower (Carthamus tinctorius L.) were subjected to three treatment durations (3h, 5h and 7h) of 0.5 % Ethyl Methane Sulphonate (EMS). Microsporogenesis was carried out in the control as well as in the treated materials. EMS treated plants showed interesting feature of partial inter-meiocyte chromatin migration through channel formation, beak formation or direct cell fusion. Another interesting feature noticed during the study was the fusion among tetrads due to wall dissolution. The phenomenon of cytomixis was recorded at nearly all the stages of microsporogenesis connecting from a few to several meiocytes. Other abnormalities such as laggards, precocious movement, bridge and non-disjunction of chromosomes were also recorded but in very low frequencies. The phenomenon of cytomixis increased along with the increase in treatment duration of EMS. Cells with these types of cytomictic disturbances may probably result in uneven formation of gametes or zygote, heterogenous sized pollen grains or even loss of fertility in future. PMID:21950142

  17. The US DOE-EM International Program - 13004

    SciTech Connect

    Elmetti, Rosa R.; Han, Ana M.; Williams, Alice C.

    2013-07-01

    The U.S. Department of Energy (DOE) Office of Environmental Management (EM) conducts international collaboration activities in support of U.S. policies and objectives regarding the accelerated risk reduction and remediation of environmental legacy of the nations' nuclear weapons program and government sponsored nuclear energy research. The EM International Program supported out of the EM Office of the Associate Principal Deputy Assistant Secretary pursues collaborations with foreign government organizations, educational institutions and private industry to assist in identifying technologies and promote international collaborations that leverage resources and link international experience and expertise. In fiscal year (FY) 2012, the International Program awarded eight international collaborative projects for work scope spanning waste processing, groundwater and soil remediation, deactivation and decommissioning (D and D) and nuclear materials disposition initiatives to seven foreign organizations. Additionally, the International Program's scope and collaboration opportunities were expanded to include technical as well as non-technical areas. This paper will present an overview of the on-going tasks awarded in FY 2012 and an update of upcoming international activities and opportunities for expansion into FY 2013 and beyond. (authors)

  18. Improving EM&V for Energy Efficiency Programs (Fact Sheet)

    SciTech Connect

    Not Available

    2012-07-01

    This fact sheet describes the objectives of the U.S. Department of Energy Uniform Methods Project to bring consistency to energy savings calculations in U.S. energy efficiency programs. The U.S. Department of Energy (DOE) is developing a framework and a set of protocols for determining gross energy savings from energy efficiency measures and programs. The protocols represent a refinement of the body of knowledge supporting energy efficiency evaluation, measurement, and verification (EM&V) activities. They have been written by technical experts within the field and reviewed by industry experts. Current EM&V practice allows for multiple methods for calculating energy savings. These methods were developed to meet the needs of energy efficiency program administrators and regulators. Although they served their original objectives well, they have resulted in inconsistent and incomparable savings results - even for identical measures. The goal of the Uniform Methods Project is to strengthen the credibility of energy savings determinations by improving EM&V, increasing the consistency and transparency of how energy savings are determined.

  19. Conjoined Use of EM and NMR in RNA Structure Refinement

    PubMed Central

    Gong, Zhou; Schwieters, Charles D.; Tang, Chun

    2015-01-01

    More than 40% of the RNA structures have been determined using nuclear magnetic resonance (NMR) technique. NMR mainly provides local structural information of protons and works most effectively on relatively small biomacromolecules. Hence structural characterization of large RNAs can be difficult for NMR alone. Electron microscopy (EM) provides global shape information of macromolecules at nanometer resolution, which should be complementary to NMR for RNA structure determination. Here we developed a new energy term in Xplor-NIH against the density map obtained by EM. We conjointly used NMR and map restraints for the structure refinement of three RNA systems — U2/U6 small-nuclear RNA, genome-packing motif (ΨCD)2 from Moloney murine leukemia virus, and ribosome-binding element from turnip crinkle virus. In all three systems, we showed that the incorporation of a map restraint, either experimental or generated from known PDB structure, greatly improves structural precision and accuracy. Importantly, our method does not rely on an initial model assembled from RNA duplexes, and allows full torsional freedom for each nucleotide in the torsion angle simulated annealing refinement. As increasing number of macromolecules can be characterized by both NMR and EM, the marriage between the two techniques would enable better characterization of RNA three-dimensional structures. PMID:25798848

  20. Minimum-distortion isometric shape correspondence using EM algorithm.

    PubMed

    Sahillioğlu, Yusuf; Yemez, Yücel

    2012-11-01

    We present a purely isometric method that establishes 3D correspondence between two (nearly) isometric shapes. Our method evenly samples high-curvature vertices from the given mesh representations, and then seeks an injective mapping from one vertex set to the other that minimizes the isometric distortion. We formulate the problem of shape correspondence as combinatorial optimization over the domain of all possible mappings, which then reduces in a probabilistic setting to a log-likelihood maximization problem that we solve via the Expectation-Maximization (EM) algorithm. The EM algorithm is initialized in the spectral domain by transforming the sampled vertices via classical Multidimensional Scaling (MDS). Minimization of the isometric distortion, and hence maximization of the log-likelihood function, is then achieved in the original 3D euclidean space, for each iteration of the EM algorithm, in two steps: by first using bipartite perfect matching, and then a greedy optimization algorithm. The optimal mapping obtained at convergence can be one-to-one or many-to-one upon choice. We demonstrate the performance of our method on various isometric (or nearly isometric) pairs of shapes for some of which the ground-truth correspondence is available.

  1. SubspaceEM: A Fast Maximum-a-posteriori Algorithm for Cryo-EM Single Particle Reconstruction

    PubMed Central

    Dvornek, Nicha C.; Sigworth, Fred J.; Tagare, Hemant D.

    2015-01-01

    Single particle reconstruction methods based on the maximum-likelihood principle and the expectation-maximization (E–M) algorithm are popular because of their ability to produce high resolution structures. However, these algorithms are computationally very expensive, requiring a network of computational servers. To overcome this computational bottleneck, we propose a new mathematical framework for accelerating maximum-likelihood reconstructions. The speedup is by orders of magnitude and the proposed algorithm produces similar quality reconstructions compared to the standard maximum-likelihood formulation. Our approach uses subspace approximations of the cryo-electron microscopy (cryo-EM) data and projection images, greatly reducing the number of image transformations and comparisons that are computed. Experiments using simulated and actual cryo-EM data show that speedup in overall execution time compared to traditional maximum-likelihood reconstruction reaches factors of over 300. PMID:25839831

  2. Inhomogeneous Media 3D EM Modeling with Integral Equation Method

    NASA Astrophysics Data System (ADS)

    di, Q.; Wang, R.; An, Z.; Fu, C.; Xu, C.

    2010-12-01

    In general, only the half space of earth is considered in electromagnetic exploration. However, for the long bipole source, because the length is close to the height of ionosphere and also most offsets between source and receivers are equal or larger than the height of ionosphere, the effect of ionosphere on the electromagnetic (EM) field should be considered when observation is carried at a very far (about several thousands kilometers) location away from the source. At this point the problem becomes one which should contain ionosphere, atmosphere and earth that is “earth-ionosphere” case. There are a few of literatures to report the electromagnetic field results which is including ionosphere, atmosphere and earth media at the same time. We firstly calculate the electromagnetic fields with the traditional controlled source (CSEM) configuration using integral equation (IE) method for a three layers earth-ionosphere model. The modeling results agree well with the half space analytical results because the effect of ionosphere for this small scale bipole source can be ignorable. The comparison of small scale three layers earth-ionosphere modeling and half space analytical resolution shows that the IE method can be used to modeling the EM fields for long bipole large offset configuration. In order to discuss EM fields’ characteristics for complicate earth-ionosphere media excited by long bipole source in the far-field and wave-guide zones, we first modeled the decay characters of electromagnetic fields for three layers earth-ionosphere model. Because of the effect of ionosphere, the earth-ionosphere electromagnetic fields’ decay curves with given frequency show that there should be an extra wave guide zone for long bipole artificial source, and there are many different characters between this extra zone and far field zone. They are: 1) the amplitudes of EM fields decay much slower; 2) the polarization patterns change; 3) the positions better to measure Zxy and

  3. Modeling of MHD edge containment in strip casting with ELEKTRA and CaPS-EM codes

    SciTech Connect

    Chang, F. C.

    2000-01-12

    This paper presents modeling studies of magnetohydrodynamics analysis in twin-roll casting. Argonne National Laboratory (ANL) and ISPAT Inland Inc. (Inland), formerly Inland Steel Co., have worked together to develop a three-dimensional (3-D) computer model that can predict eddy currents, fluid flows, and liquid metal containment of an electromagnetic (EM) edge containment device. The model was verified by comparing predictions with experimental results of liquid metal containment and fluid flow in EM edge dams (EMDs) that were designed at Inland for twin-roll casting. This mathematical model can significantly shorten casting research on the use of EM fields for liquid metal containment and control. The model can optimize the EMD design so it is suitable for application, and minimize expensive time-consuming full-scale testing. Numerical simulation was performed by coupling a 3-D finite-element EM code (ELEKTRA) and a 3-D finite-difference fluids code (CaPS-EM) to solve heat transfer, fluid flow, and turbulence transport in a casting process that involves EM fields. ELEKTRA can predict the eddy-current distribution and the EM forces in complex geometries. CaPS-EM can model fluid flows with free surfaces. The computed 3-D magnetic fields and induced eddy currents in ELEKTRA are used as input to temperature- and flow-field computations in CaPS-EM. Results of the numerical simulation compared well with measurements obtained from both static and dynamic tests.

  4. Emergency medical services and "psych calls": Examining the work of urban EMS providers.

    PubMed

    Prener, Christopher; Lincoln, Alisa K

    2015-11-01

    Emergency medical technicians and paramedics form the backbone of the United States' Emergency Medical Service (EMS) system. Despite the frequent involvement of EMS with people with mental health and substance abuse problems, the nature and content of this work, as well as how EMS providers think about this work, have not been fully explored. Using data obtained through observations and interviews with providers at an urban American EMS agency, this paper provides an analysis of the ways in which EMS providers interact with people with mental illness and substance abuse problems, as well as providers' experiences with the mental health care system. Results demonstrate that EMS providers share common beliefs and frustrations about "psych calls" and the types of calls that involve people with behavioral health problems. In addition, providers described their understandings of the ways in which people with mental health and substance use problems "abuse the system" and the consequences of this abuse. Finally, EMS providers discuss the system-level factors that impact their work and specific barriers and challenges to care. These results suggest that additional work is needed to expand our understanding of the role of EMS providers in the care of people with behavioral health problems and that mental health practitioners and policy makers should include consideration of the important role of EMS and prehospital care in providing community-based supports for people with behavioral health needs. (PsycINFO Database Record

  5. Volta phase plate cryo-EM of the small protein complex Prx3

    NASA Astrophysics Data System (ADS)

    Khoshouei, Maryam; Radjainia, Mazdak; Phillips, Amy J.; Gerrard, Juliet A.; Mitra, Alok K.; Plitzko, Jürgen M.; Baumeister, Wolfgang; Danev, Radostin

    2016-01-01

    Cryo-EM of large, macromolecular assemblies has seen a significant increase in the numbers of high-resolution structures since the arrival of direct electron detectors. However, sub-nanometre resolution cryo-EM structures are rare compared with crystal structure depositions, particularly for relatively small particles (<400 kDa). Here we demonstrate the benefits of Volta phase plates for single-particle analysis by time-efficient cryo-EM structure determination of 257 kDa human peroxiredoxin-3 dodecamers at 4.4 Å resolution. The Volta phase plate improves the applicability of cryo-EM for small molecules and accelerates structure determination.

  6. Perda de massa em ventos empoeirados de estrelas supergigantes

    NASA Astrophysics Data System (ADS)

    Vidotto, A. A.; Jatenco-Pereira, V.

    2003-08-01

    Em praticamente todas as regiões do diagrama HR, as estrelas apresentam evidências observacionais de perda de massa. Na literatura, pode-se encontrar trabalhos que tratam tanto do diagnóstico da perda de massa como da construção de modelos que visam explicá-la. O amortecimento de ondas Alfvén tem sido utilizado como mecanismo de aceleração de ventos homogêneos. Entretanto, sabe-se que os envelopes de estrelas frias contêm grãos sólidos e moléculas. Com o intuito de estudar a interação entre as ondas Alfvén e a poeira e a sua conseqüência na aceleração do vento estelar, Falceta-Gonçalves & Jatenco-Pereira (2002) desenvolveram um modelo de perda de massa para estrelas supergigantes. Neste trabalho, apresentamos um estudo do modelo acima proposto para avaliar a dependência da taxa de perda de massa com alguns parâmetros iniciais como, por exemplo, a densidade r0, o campo magnético B0, o comprimento de amortecimento da onda L0, seu fluxo f0, entre outros. Sendo assim, aumentando f0 de 10% a partir de valores de referência, vimos que aumenta consideravelmente, enquanto que um aumento de mesmo valor em r0, B0 e L0 acarreta uma diminuição em .

  7. EM-Based Multiuser Detection in Fast Fading Multipath Environments

    NASA Astrophysics Data System (ADS)

    Borran, Mohammad Jaber; Aazhang, Behnaam

    2002-12-01

    We address the problem of multiuser detection in fast fading multipath environments for DS-CDMA systems. In fast fading scenarios, temporal variations of the channel cause significant performance degradation even with the Rake receiver. We use a previously introduced time-frequency (TF) Rake receiver based on a canonical formulation of the channel and signals to simultaneously combat fading and multipath effects. This receiver uses the Doppler spread caused by rapid time-varying channel as another means of diversity. In dealing with multiaccess interference and as an attempt to avoid the prohibitive computational complexity of the optimum maximum-likelihood (ML) detector, we use the expectation maximization (EM) algorithm to derive an approximate ML detector. The new detector turns out to have an iterative structure very similar to the well-known multistage detector with some extra parameters. At the two extreme values of these parameters, the EM detector reduces to either one-shot TF Rake or generalized multistage detector. For the intermediate values of the parameters, it combines the two estimates to obtain a better decision for the bits of the users. Because of using the EM algorithm, this detector has better convergence properties than the multistage detector; the bit estimates always converge, and if an appropriate initial vector is used, they converge to the global maximizer of the likelihood function. As a result, the new detector provides significantly improved performance while maintaining the low complexity of the multistage detector. Our simulation results confirm the expected performance improvements compared to the base case of the TF Rake as well as the multistage detector used with the TF Rake.

  8. EM-21 Retrieval Knowledge Center: Waste Retrieval Challenges

    SciTech Connect

    Fellinger, Andrew P.; Rinker, Michael W.; Berglin, Eric J.; Minichan, Richard L.; Poirier, Micheal R.; Gauglitz, Phillip A.; Martin, Bruce A.; Hatchell, Brian K.; Saldivar, Eloy; Mullen, O Dennis; Chapman, Noel F.; Wells, Beric E.; Gibbons, Peter W.

    2009-04-10

    EM-21 is the Waste Processing Division of the Office of Engineering and Technology, within the U.S. Department of Energy’s (DOE) Office of Environmental Management (EM). In August of 2008, EM-21 began an initiative to develop a Retrieval Knowledge Center (RKC) to provide the DOE, high level waste retrieval operators, and technology developers with centralized and focused location to share knowledge and expertise that will be used to address retrieval challenges across the DOE complex. The RKC is also designed to facilitate information sharing across the DOE Waste Site Complex through workshops, and a searchable database of waste retrieval technology information. The database may be used to research effective technology approaches for specific retrieval tasks and to take advantage of the lessons learned from previous operations. It is also expected to be effective for remaining current with state-of-the-art of retrieval technologies and ongoing development within the DOE Complex. To encourage collaboration of DOE sites with waste retrieval issues, the RKC team is co-led by the Savannah River National Laboratory (SRNL) and the Pacific Northwest National Laboratory (PNNL). Two RKC workshops were held in the Fall of 2008. The purpose of these workshops was to define top level waste retrieval functional areas, exchange lessons learned, and develop a path forward to support a strategic business plan focused on technology needs for retrieval. The primary participants involved in these workshops included retrieval personnel and laboratory staff that are associated with Hanford and Savannah River Sites since the majority of remaining DOE waste tanks are located at these sites. This report summarizes and documents the results of the initial RKC workshops. Technology challenges identified from these workshops and presented here are expected to be a key component to defining future RKC-directed tasks designed to facilitate tank waste retrieval solutions.

  9. Covariance Matrix Estimation for the Cryo-EM Heterogeneity Problem*

    PubMed Central

    Katsevich, E.; Katsevich, A.; Singer, A.

    2015-01-01

    In cryo-electron microscopy (cryo-EM), a microscope generates a top view of a sample of randomly oriented copies of a molecule. The problem of single particle reconstruction (SPR) from cryo-EM is to use the resulting set of noisy two-dimensional projection images taken at unknown directions to reconstruct the three-dimensional (3D) structure of the molecule. In some situations, the molecule under examination exhibits structural variability, which poses a fundamental challenge in SPR. The heterogeneity problem is the task of mapping the space of conformational states of a molecule. It has been previously suggested that the leading eigenvectors of the covariance matrix of the 3D molecules can be used to solve the heterogeneity problem. Estimating the covariance matrix is challenging, since only projections of the molecules are observed, but not the molecules themselves. In this paper, we formulate a general problem of covariance estimation from noisy projections of samples. This problem has intimate connections with matrix completion problems and high-dimensional principal component analysis. We propose an estimator and prove its consistency. When there are finitely many heterogeneity classes, the spectrum of the estimated covariance matrix reveals the number of classes. The estimator can be found as the solution to a certain linear system. In the cryo-EM case, the linear operator to be inverted, which we term the projection covariance transform, is an important object in covariance estimation for tomographic problems involving structural variation. Inverting it involves applying a filter akin to the ramp filter in tomography. We design a basis in which this linear operator is sparse and thus can be tractably inverted despite its large size. We demonstrate via numerical experiments on synthetic datasets the robustness of our algorithm to high levels of noise. PMID:25699132

  10. Epidemiology of major incidents: an EMS study from Pakistan

    PubMed Central

    2011-01-01

    Background A major incident is defined as an event that owing to the number of casualties has the potential to overwhelm the available resources. This paper attempts to describe the incidence and epidemiology of major incidents dealt with by a government-run emergency medical service (EMS) in the Punjab province of Pakistan, a developing country in South Asia. A major incident in this EMS is defined as any incident that produces three or more patients, or any incident in which extraordinary resources are needed. Methods All the calls received by an EMS Rescue 1122 were studied over a 6-month period. Calls that were defined as major incidents were identified, and further details were sought from the districts regarding these incidents. Questions specifically asked were the type of incident, time of the incident, response time for the incident, the resources needed, and the number of dead and injured casualties. Retrospective data were collected from the submitted written reports. Results Road traffic crashes (RTCs) emerged as the leading cause of a major incident in the province of Punjab and also led to the greatest number of casualties, followed by fire incidents. The total number of casualties was 3,380, out of which 73.7% were RTC victims. There was a high rate of death on the scene (10.4%). Certain other causes of major incidents also emerged, including violence, gas explosions and drowning. Conclusion Road traffic crashes are the most common cause of a major incident in developing countries such as Pakistan. Injury prevention initiatives need to focus on RTCs. PMID:21798011

  11. The effective molarity (EM) puzzle in proton transfer reactions.

    PubMed

    Karaman, Rafik

    2009-08-01

    The DFT and HF calculation results for the proton transfer reactions of three different systems reveal that the reaction mechanism (transfer of a proton to a nucleophile) is largely determined by the distance between the two reactive centers (r). Systems with relatively large r values tend to abstract a proton from a molecule of water, whereas, these with a relatively small r values prefer to be engaged intramolecularly and their interaction with water is only via hydrogen bonding. Further, the results indicate that the effective molarity (logEM) for an intramolecular process is strongly correlated with the distance between the two reacting centers (r) in accordance with Menger's "spatiotemporal hypothesis".

  12. EMS Bill of Rights: what every patient deserves.

    PubMed

    Meador, Steven R; Slovis, Corey M; Wrenn, Keith D

    2003-03-01

    Every EMT, paramedic, supervisor and EMS medical director must stress that our job is to treat every patient the way we'd like to be treated or the way we'd want one of our family members treated. Those charged with supervision must ensure that EMTs and paramedics receive the support and training necessary to allow adherence to the 10 objectives outlined in this article. Quality-assurance reviews, quality-improvement programs, provider comments and complaints from the public must all be used to improve the care we deliver. Prehospital care should be constantly improving the delivery of excellent, life-saving care.

  13. On the EM algorithm for overdispersed count data.

    PubMed

    McLachlan, G J

    1997-03-01

    In this paper, we consider the use of the EM algorithm for the fitting of distributions by maximum likelihood to overdispersed count data. In the course of this, we also provide a review of various approaches that have been proposed for the analysis of such data. As the Poisson and binomial regression models, which are often adopted in the first instance for these analyses, are particular examples of a generalized linear model (GLM), the focus of the account is on the modifications and extensions to GLMs for the handling of overdispersed count data. PMID:9185291

  14. Theme park EMS. It's no Mickey Mouse operation.

    PubMed

    Philips, J H

    1995-07-01

    In a single shift, they can rescue someone trapped in a time warp, perform CPR while menacing monsters look over their shoulders or calmly watch as a motorboat chase ends in a huge ball of fire. It's just another routine day for EMS providers in America's theme parks, where getting ready for unusual calls is the norm. JEMS went behind the scenes with theme park responders to explore the unique challenges of their service environment. What we found was a group of super-dedicated professionals who place genuine hospitality right alongside quality patient care. PMID:10143711

  15. Single-particle cryo-EM at crystallographic resolution

    PubMed Central

    Cheng, Yifan

    2015-01-01

    Until only a few years ago, single-particle electron cryo-microscopy (cryo-EM) was usually not the first choice for many structural biologists due to its limited resolution in the range of nanometer to subnanometer. Now, this method rivals X-ray crystallography in terms of resolution and can be used to determine atomic structures of macromolecules that are either refractory to crystallization or difficult to crystallize in specific functional states. In this review, I discuss the recent breakthroughs in both hardware and software that transformed cryo-microscopy, enabling understanding of complex biomolecules and their functions at atomic level. PMID:25910205

  16. State of the Art in EM Field Computation

    SciTech Connect

    Ng, C.; Akcelik, V.; Candel, A.; Chen, S.; Folwell, N.; Ge, L.; Guetz, A.; Jiang, H.; Kabel, A.; Lee, L.-Q.; Li, Z.; Prudencio, E.; Schussman, G.; Uplenchwar, R.; Xiao, L.; Ko, K.; /SLAC

    2006-09-25

    This paper presents the advances in electromagnetic (EM) field computation that have been enabled by the US DOE SciDAC Accelerator Science and Technology project which supports the development and application of a suite of electromagnetic codes based on the higher-order finite element method. Implemented on distributed memory supercomputers, this state of the art simulation capability has produced results which are of great interest to accelerator designers and with realism previously not possible with standard codes. Examples from work on the International Linear Collider (ILC) project are described.

  17. Covariance Structure Model Fit Testing under Missing Data: An Application of the Supplemented EM Algorithm

    ERIC Educational Resources Information Center

    Cai, Li; Lee, Taehun

    2009-01-01

    We apply the Supplemented EM algorithm (Meng & Rubin, 1991) to address a chronic problem with the "two-stage" fitting of covariance structure models in the presence of ignorable missing data: the lack of an asymptotically chi-square distributed goodness-of-fit statistic. We show that the Supplemented EM algorithm provides a convenient…

  18. Method for evaluating compatibility of commercial electromagnetic (EM) microsensor tracking systems with surgical and imaging tables

    NASA Astrophysics Data System (ADS)

    Nafis, Christopher; Jensen, Vern; von Jako, Ron

    2008-03-01

    Electromagnetic (EM) tracking systems have been successfully used for Surgical Navigation in ENT, cranial, and spine applications for several years. Catheter sized micro EM sensors have also been used in tightly controlled cardiac mapping and pulmonary applications. EM systems have the benefit over optical navigation systems of not requiring a line-of-sight between devices. Ferrous metals or conductive materials that are transient within the EM working volume may impact tracking performance. Effective methods for detecting and reporting EM field distortions are generally well known. Distortion compensation can be achieved for objects that have a static spatial relationship to a tracking sensor. New commercially available micro EM tracking systems offer opportunities for expanded image-guided navigation procedures. It is important to know and understand how well these systems perform with different surgical tables and ancillary equipment. By their design and intended use, micro EM sensors will be located at the distal tip of tracked devices and therefore be in closer proximity to the tables. Our goal was to define a simple and portable process that could be used to estimate the EM tracker accuracy, and to vet a large number of popular general surgery and imaging tables that are used in the United States and abroad.

  19. Global Convergence of the EM Algorithm for Unconstrained Latent Variable Models with Categorical Indicators

    ERIC Educational Resources Information Center

    Weissman, Alexander

    2013-01-01

    Convergence of the expectation-maximization (EM) algorithm to a global optimum of the marginal log likelihood function for unconstrained latent variable models with categorical indicators is presented. The sufficient conditions under which global convergence of the EM algorithm is attainable are provided in an information-theoretic context by…

  20. Developing State and National Evaluation Infrastructures- Guidance for the Challenges and Opportunities of EM&V

    SciTech Connect

    Schiller, Steven R.; Goldman, Charles A.

    2011-06-24

    Evaluating the impacts and effectiveness of energy efficiency programs is likely to become increasingly important for state policymakers and program administrators given legislative mandates and regulatory goals and increasing reliance on energy efficiency as a resource. In this paper, we summarize three activities that the authors have conducted that highlight the expanded role of evaluation, measurement and verification (EM&V): a study that identified and analyzed challenges in improving and scaling up EM&V activities; a scoping study that identified issues involved in developing a national efficiency EM&V standard; and lessons learned from providing technical assistance on EM&V issues to states that are ramping up energy efficiency programs. The lessons learned are summarized in 13 EM&V issues that policy makers should address in each jurisdiction and which are listed and briefly described. The paper also discusses how improving the effectiveness and reliability of EM&V will require additional capacity building, better access to existing EM&V resources, new methods to address emerging issues and technologies, and perhaps foundational documents and approaches to improving the credibility and cross jurisdictional comparability of efficiency investments. Two of the potential foundational documents discussed are a national EM&V standard or resource guide and regional deemed savings and algorithm databases.

  1. Fabrication and EM shielding properties of electrospining PANi/MWCNT/PEO fibrous membrane and its composite

    NASA Astrophysics Data System (ADS)

    Zhang, Zhichun; Jiang, Xueyong; Liu, Yanju; Leng, Jinsong

    2012-04-01

    In this paper, Polyaniline-based fibrous membranes were fabricated with multi-walled carbon nanotubes and polyethylene oxide (PEO) by the electrospinning method. And then PANi/PEO/MWCNT fibrous membranes reinforced epoxy based nanocomposite was then fabricated. The morphology and electrical properties of PANi /MWCNT /PEO fibrous membrane was characterized by scanning electron microscope (SEM). The morphologies of the membranes indicate that the electrospining method can fabricate well nano structures fibrous membrane. The EM properties of the composite reinforced with the electrospining fibrous membrane were measured by vector network analyzer. The results show that the permittivity real, image parts and permeability real part of the composite increase by filling with PANI/PEO and PANI/CNT/PEO membrane. The EM shielding and absorb performance is base on the dielectric dissipation. And different membranes made of different materials show different EM parameter, and different EM shielding performance, which can be used to the EM shielding and stealth material design and fabrication.

  2. Data-adaptive Shrinkage via the Hyperpenalized EM Algorithm

    PubMed Central

    Boonstra, Philip S.; Taylor, Jeremy M. G.; Mukherjee, Bhramar

    2015-01-01

    We propose an extension of the expectation-maximization (EM) algorithm, called the hyperpenalized EM (HEM) algorithm, that maximizes a penalized log-likelihood, for which some data are missing or unavailable, using a data-adaptive estimate of the penalty parameter. This is potentially useful in applications for which the analyst is unable or unwilling to choose a single value of a penalty parameter but instead can posit a plausible range of values. The HEM algorithm is conceptually straightforward and also very effective, and we demonstrate its utility in the analysis of a genomic data set. Gene expression measurements and clinical covariates were used to predict survival time. However, many survival times are censored, and some observations only contain expression measurements derived from a different assay, which together constitute a difficult missing data problem. It is desired to shrink the genomic contribution in a data-adaptive way. The HEM algorithm successfully handles both the missing data and shrinkage aspects of the problem. PMID:26834856

  3. The Search for Gravitational Wave EM Counterparts with Swift

    NASA Astrophysics Data System (ADS)

    Kennea, Jamie; Evans, Phil; Swift GW follow-up Team

    2016-04-01

    We present the plan to search for electromagnetic counterparts of Gravitational Waves (GWs) discovered during the current and upcoming runs of the LIGO and Virgo detectors. As we enter a period where the sensitivity of the current generation of GW detectors approaches a high probability of the first detection of a real GW signal, confirmation of the reality of these triggers will be greatly improved if an EM counterpart can be found. Swift’s ability to rapidly respond to high priority target-of-opportunity observations, it’s multi-wavelength capabilities and low overhead observing make it a seemingly ideal follow-up facility. However comparing the size of the expected GW error regions with the fields of view of the Swift XRT and UVOT telescopes, we find that covering the large GW error regions would require a unreasonably large number of pointings. We present our method of meeting this challenge, by both reducing the problem using Galaxy targeting, and by operating Swift in an entirely new way in order to cover the still large number of fields needed to chase down the EM counterpart before it disappears.

  4. Chapter 19: HVAC Controls (DDC/EMS/BAS) Evaluation Protocol

    SciTech Connect

    Romberger, J.

    2014-11-01

    The HVAC Controls Evaluation Protocol is designed to address evaluation issues for direct digital controls/energy management systems/building automation systems (DDC/EMS/BAS) that are installed to control heating, ventilation, and air-conditioning (HVAC) equipment in commercial and institutional buildings. (This chapter refers to the DDC/EMS/BAS measure as HVAC controls.) This protocol may also be applicable to industrial facilities such as clean rooms and labs, which have either significant HVAC equipment or spaces requiring special environmental conditions. This protocol addresses only HVAC-related equipment and the energy savings estimation methods associated with installing such control systems as an energy efficiency measure. The affected equipment includes: Air-side equipment (air handlers, direct expansion systems, furnaces, other heating- and cooling-related devices, terminal air distribution equipment, and fans); Central plant equipment (chillers, cooling towers, boilers, and pumps). These controls may also operate or affect other end uses, such as lighting, domestic hot water, irrigation systems, and life safety systems such as fire alarms and other security systems. Considerable nonenergy benefits, such as maintenance scheduling, system component troubleshooting, equipment failure alarms, and increased equipment lifetime, may also be associated with these systems. When connected to building utility meters, these systems can also be valuable demand-limiting control tools. However, this protocol does not evaluate any of these additional capabilities and benefits.

  5. Contactless ultrasonic treatment of melts using EM induction

    NASA Astrophysics Data System (ADS)

    Bojarevics, V.; Djambazov, G.; Lebon, G. S. B.; Pericleous, K. A.

    2015-06-01

    Ultrasound Treatment (UT) is commonly used in light alloys during solidification to refine microstructure, or disperse immersed particles. A sonotrode probe introduced into the melt generates sound waves that are strong enough to produce cavitation of dissolved gases. The same method cannot be used in high temperature melts, or for highly reactive alloys, due to probe erosion and melt contamination. An alternative, contactless method of generating sound waves is proposed and investigated theoretically in this paper, using electromagnetic (EM) induction. In addition to strong vibration, the EM induction currents generate strong stirring in the melt that aids distribution of the UT effect to large volumes of material. In a typical application, the same induction coil surrounding the crucible used to melt the alloy may be adopted for UT with suitable frequency tuning. Alternatively - or in addition - a top coil may be used. For industrial use, instead of multiple sonotrodes as has been the practice in scaling up, modelling shows that one simply has to alter the coil geometry and current to suit. To reach sinusoidal pressure fluctuations suitable for cavitation it may be necessary to tune the induction coil frequency for resonance, given the crucible dimensions.

  6. The EM fields in the Solid Generated by a Fault in a Porous Region

    NASA Astrophysics Data System (ADS)

    Ren, H.; Huang, Q.; Chen, X.

    2015-12-01

    Electrokinetic effect, as one of the most possible generation mechanisms of the seismo-electromagnetic phenomenons associated with natural earthquakes, has interested many researchers. Besides, it is also considered as a potential tool for the water/oil exploration. Recently, we numerically investigated the electromagnetic (EM) fields due to the electrokinetic effect in mixed layered model. The mixed model comprises not only porous layers but also solid layers. We firstly tested a two-layer mixed model. The numerical results show that, in addition to the radiation EM fields, another kind of evanescent EM fields can be generated by the seismic waves arriving at the interface with incident angles greater the critical angle. The evanescent EM fields decay faster than the radiation EM fields when getting away from the interface. For the seismic frequency band, the evanescent EM fields in the solid are still measurable at a distance of, e.g., 2km to the interface. We then tested a eight-layer mixed model. The top and bottom layers are solid and the other layers are porous. A finite fault of 20x10km is located in the porous region. The focal depth is 8km. The applied source time function is a ramp fuction with an arise time of 0.8s. Point stacking method was used to compute the wave-fields caused by the finite fault. Our nuemrical results show that, this model can generate the EM fields before the arrival of seismic waves as well as the residual EM fields. Both the two kinds of EM fields have been observed in field observations. There is a possibility that the anomalous EM activities before big earthquakes may be caused by the fluid flow in the shallow Earth as a result of the stress changes.

  7. Estudo em microondas do aprisionamento e precipitação de elétrons em explosões solares

    NASA Astrophysics Data System (ADS)

    Rosal, A. C.; Costa, J. E. R.

    2003-08-01

    Uma explosão solar é uma variação rápida e intensa do brilho que ocorre nas chamadas regiões ativas da atmosfera, constituídas por um plasma magnetizado com intensa indução magnética. Os modelos de explosões solares atuais, discutidos na literatura, apresentam características de aprisionamento e precipitação de elétrons em ambientes magnéticos simplificados. Neste trabalho, nos propusemos a separar a emissão dos elétrons aprisionados da emissão dos elétrons em precipitação apenas a partir da emissão em microondas, melhorando portanto o controle sobre o conjunto de parâmetros inferidos. A emissão em microondas da população em precipitação é bastante fraca e portanto da nossa base de dados de 130 explosões observadas pelo Rádio Polarímetro de Nobeyama, em sete freqüências, apenas para 32 foi possível separar as duas componentes de emissão com uma boa razão sinal/ruído. A partir de estudos das escalas de tempo das emissões devidas à variação gradual da emissão no aprisionamento e da variação rápida da emissão dos elétrons em precipitação foi possível obter a separação utilizando um filtro temporal nas emissões resultantes. Em nossa análise destas explosões estudamos os espectros girossincrotrônicos da emissão gradual, a qual associamos provir do topo dos arcos magnéticos e da emissão de variação rápida associada aos elétrons em precipitação. Estes espectros foram calculados e dos quais inferimos que a indução magnética efetiva do topo e dos pés foi em média, Btopo = 236 G e Bpés = 577 G, inferidas das freqüências de pico dos espectros em ntopo = 11,8 GHz e npés = 14,6 GHz com leve anisotropia (pequeno alargamento espectral). O índice espectral da distribuição não-térmica de elétrons d, inferido do índice espectral de fótons da emissão em regime opticamente fino, foi de dtopo = 3,3 e dpés = 3,9. Estes parâmetros são típicos da maioria das análises realizadas em ambiente único de

  8. The EM SSAB Annual Work Plan Process: Focusing Board Efforts and Resources - 13667

    SciTech Connect

    Young, Ralph

    2013-07-01

    One of the most daunting tasks for any new member of a local board of the Environmental Management Site Specific Advisory Board (EM SSAB) is to try to understand the scope of the clean-up activities going on at the site. In most cases, there are at least two or three major cleanup activities in progress as well as monitoring of past projects. When planning for future projects is added to the mix, the list of projects can be long. With the clean-up activities involving all major environmental media - air, water, soils, and groundwater, new EM SSAB members can find themselves totally overwhelmed and ineffective. Helping new members get over this initial hurdle is a major objective of EM and all local boards of the EM SSAB. Even as members start to understand the size and scope of the projects at a site, they can still be frustrated at the length of time it takes to see results and get projects completed. Many project and clean-up timelines for most of the sites go beyond 10 years, so it's not unusual for an EM SSAB member to see the completion of only 1 or 2 projects over the course of their 6-year term on the board. This paper explores the annual work planning process of the EM SSAB local boards, one tool that can be used to educate EM SSAB members into seeing the broader picture for the site. EM SSAB local work plans divide the site into projects focused on a specific environmental issue or media such as groundwater and/or waste disposal options. Projects are further broken down into smaller segments by highlighting major milestones. Using these metrics, local boards of the EM SSAB can start to quantify the effectiveness of the project in achieving the ultimate goal of site clean-up. These metrics can also trigger board advice and recommendations for EM. At the beginning of each fiscal year, the EM SSAB work plan provides a road map with quantifiable checkpoints for activities throughout the year. When the work plans are integrated with site-specific, enforceable

  9. EM Structure-Based Accelerators Working Group Summary

    SciTech Connect

    Kimura, W.D.; Lidia, S.M.

    2004-12-07

    This Working Group (WG) focused on EM Structure-Based Accelerators, which covers a broad area of mechanisms and experiments. Topics covered included dielectric wakefield accelerators (DWA), photonic bandgap accelerators (PBGA), inverse free electron lasers (IFEL), vacuum laser accelerators (VLA), other novel schemes, and supporting analysis and modeling. In addition, this WG was tasked at the Workshop with developing conceptual (strawman) designs for a 1-GeV accelerator system based upon any of the experimentally-proven approaches covered in this WG. Two strawmen designs were developed based upon IFELs and DWAs. The presentations given and strawmen designs indicate great progress has been made in many areas. Proof-of-principle experiments will occur shortly in PBGA and VLA. Other well-proven devices, such as IFELs, are becoming accepted as 'workhorse' providers of microbunches.

  10. Kaisen. EMS as theater of the streets. Part two.

    PubMed

    Dernocoeur, K; Taigman, M

    1991-03-01

    Passing an EMT or paramedic course is a considerable achievement, as you have had to acquire a huge and fascinating volume of medical knowledge. But, did the program teach you acting skills? Acting? Whoever said that EMS professionals need to know anything about acting? We submit that you do; prehospital workers can gain many unanticipated benefits by using various acting skills and tactics in the field. One such benefit is the avoidance of the "complacency trap," or the boredom that can develop if you adopt the viewpoint that one crisis is much like another. But, more importantly, you will be more effective at delivering prehospital care, and you'll have a lot more fun on the job.

  11. Kaisen. EMS as theater of the streets. Part two.

    PubMed

    Dernocoeur, K; Taigman, M

    1991-03-01

    Passing an EMT or paramedic course is a considerable achievement, as you have had to acquire a huge and fascinating volume of medical knowledge. But, did the program teach you acting skills? Acting? Whoever said that EMS professionals need to know anything about acting? We submit that you do; prehospital workers can gain many unanticipated benefits by using various acting skills and tactics in the field. One such benefit is the avoidance of the "complacency trap," or the boredom that can develop if you adopt the viewpoint that one crisis is much like another. But, more importantly, you will be more effective at delivering prehospital care, and you'll have a lot more fun on the job. PMID:10110186

  12. Three penalized EM-type algorithms for PET image reconstruction.

    PubMed

    Teng, Yueyang; Zhang, Tie

    2012-06-01

    Based on Bayes theory, Green introduced the maximum a posteriori (MAP) algorithm to obtain a smoothing reconstruction for positron emission tomography. This algorithm is flexible and convenient for most of the penalties, but it is hard to guarantee convergence. For a common goal, Fessler penalized a weighted least squares (WLS) estimator by a quadratic penalty and then solved it with the successive over-relaxation (SOR) algorithm, however, the algorithm was time-consuming and difficultly parallelized. Anderson proposed another WLS estimator for faster convergence, on which there were few regularization methods studied. For three regularized estimators above, we develop three new expectation maximization (EM) type algorithms to solve them. Unlike MAP and SOR, the proposed algorithms yield update rules by minimizing the auxiliary functions constructed on the previous iterations, which ensure the cost functions monotonically decreasing. Experimental results demonstrated the robustness and effectiveness of the proposed algorithms.

  13. Telescópio de patrulhamento solar em 12 GHz

    NASA Astrophysics Data System (ADS)

    Utsumi, F.; Costa, J. E. R.

    2003-08-01

    O telescópio de patrulhamento solar é um instrumento dedicado à observação de explosões solares com início de suas operações em janeiro de 2002, trabalhando próximo ao pico de emissão do espectro girossincrotrônico (12 GHz). Trata-se de um arranjo de três antenas concebido para a detecção de explosões e determinação em tempo real da localização da região emissora. Porém, desde sua implementação em uma montagem equatorial movimentada por um sistema de rotação constante (15 graus/hora) o rastreio apresentou pequenas variações de velocidade e folgas nas caixas de engrenagens. Assim, tornou-se necessária a construção de um sistema de correção automática do apontamento que era de fundamental importância para os objetivos do projeto. No segundo semestre de 2002 empreendemos uma série de tarefas com o objetivo de automatizar completamente o rastreio, a calibração, a aquisição de dados, controle de ganhos, offsets e transferência dos dados pela internet através de um projeto custeado pela FAPESP. O rastreio automático é realizado através de um inversor que controla a freqüência da rede de alimentação do motor de rastreio podendo fazer micro-correções na direção leste-oeste conforme os radiômetros desta direção detectem uma variação relativa do sinal. Foi adicionado também um motor na direção da declinação para correção automática da variação da direção norte-sul. Após a implementação deste sistema a precisão do rastreio melhorou para um desvio máximo de 30 segundos de arco, o que está muito bom para este projeto. O Telescópio se encontra em funcionamento automático desde março de 2003 e já conta com várias explosões observadas após a conclusão desta fase de automação. Estamos apresentando as explosões mais intensas do período e com as suas respectivas posições no disco solar.

  14. Imaging protein three-dimensional nanocrystals with cryo-EM.

    PubMed

    Nederlof, Igor; Li, Yao Wang; van Heel, Marin; Abrahams, Jan Pieter

    2013-05-01

    Flash-cooled three-dimensional crystals of the small protein lysozyme with a thickness of the order of 100 nm were imaged by 300 kV cryo-EM on a Falcon direct electron detector. The images were taken close to focus and to the eye appeared devoid of contrast. Fourier transforms of the images revealed the reciprocal lattice up to 3 Å resolution in favourable cases and up to 4 Å resolution for about half the crystals. The reciprocal-lattice spots showed structure, indicating that the ordering of the crystals was not uniform. Data processing revealed details at higher than 2 Å resolution and indicated the presence of multiple mosaic blocks within the crystal which could be separately processed. The prospects for full three-dimensional structure determination by electron imaging of protein three-dimensional nanocrystals are discussed. PMID:23633595

  15. Speech articulator measurements using low power EM-wave sensors

    SciTech Connect

    Holzrichter, J.F.; Burnett, G.C.; Ng, L.C.; Lea, W.A.

    1998-01-01

    Very low power electromagnetic (EM) wave sensors are being used to measure speech articulator motions as speech is produced. Glottal tissue oscillations, jaw, tongue, soft palate, and other organs have been measured. Previously, microwave imaging (e.g., using radar sensors) appears not to have been considered for such monitoring. Glottal tissue movements detected by radar sensors correlate well with those obtained by established laboratory techniques, and have been used to estimate a voiced excitation function for speech processing applications. The noninvasive access, coupled with the small size, low power, and high resolution of these new sensors, permit promising research and development applications in speech production, communication disorders, speech recognition and related topics. {copyright} {ital 1998 Acoustical Society of America.}

  16. EM Properties of Magnetic Minerals at RADAR Frequencies

    NASA Technical Reports Server (NTRS)

    Stillman, D. E.; Olhoeft, G. R.

    2005-01-01

    Previous missions to Mars have revealed that Mars surface is magnetic at DC frequency. Does this highly magnetic surface layer attenuate RADAR energy as it does in certain locations on Earth? It has been suggested that the active magnetic mineral on Mars is titanomaghemite and/or titanomagnetite. When titanium is incorporated into a maghemite or magnetite crystal, the Curie temperature can be significantly reduced. Mars has a wide range of daily temperature fluctuations (303K - 143K), which could allow for daily passes through the Curie temperature. Hence, the global dust layer on Mars could experience widely varying magnetic properties as a function of temperature, more specifically being ferromagnetic at night and paramagnetic during the day. Measurements of EM properties of magnetic minerals were made versus frequency and temperature (300K- 180K). Magnetic minerals and Martian analog samples were gathered from a number of different locations on Earth.

  17. Detection of karst structures using airborne EM and VLF

    SciTech Connect

    Beard, L.P. Nyquist, J.E.; Carpenter, P.J.

    1994-12-31

    Through the combined use of multi-frequency helicopter electromagnetic and VLF data, it is possible to detect and delineate a wide variety of karst structures and possibly to assess their interconnectedness. Multi-frequency EM Can detect karst features if some element of the structure is conductive. This conductive aspect may derive from thick, moist soils in the depression commonly associated with a doline, from conductive fluids in the cavity, or from conductive sediments in the cavity if these occupy a significant portion of it. Multiple loop configurations may also increase the likelihood of detecting karst features. Preliminary evidence indicates total field VLF measurements may be able to detect interconnected karst pathways, so long as the pathways are water or sediment filled. Neither technique can effectively detect dry, resistive air-filled cavities.

  18. Integrated EM & Thermal Simulations with Upgraded VORPAL Software

    SciTech Connect

    D.N. Smithe, D. Karipides, P. Stoltz, G. Cheng, H. Wang

    2011-03-01

    Nuclear physics accelerators are powered by microwaves which must travel in waveguides between room-temperature sources and the cryogenic accelerator structures. The ohmic heat load from the microwaves is affected by the temperature-dependent surface resistance and in turn affects the cryogenic thermal conduction problem. Integrated EM & thermal analysis of this difficult non-linear problem is now possible with the VORPAL finite-difference time-domain simulation tool. We highlight thermal benchmarking work with a complex HOM feed-through geometry, done in collaboration with researchers at the Thomas Jefferson National Accelerator Laboratory, and discuss upcoming design studies with this emerging tool. This work is part of an effort to generalize the VORPAL framework to include generalized PDE capabilities, for wider multi-physics capabilities in the accelerator, vacuum electronics, plasma processing and fusion R&D fields, and we will also discuss user interface and algorithmic upgrades which facilitate this emerging multiphysics capability.

  19. EM modeling of RF drive in DTL tank 4

    SciTech Connect

    Kurennoy, Sergey S.

    2012-06-19

    A 3-D MicroWave Studio model for the RF drive in the LANSCE DTL tank 4 has been built. Both eigensolver and time-domain modeling are used to evaluate maximal fields in the drive module and RF coupling. The LANSCE DTL tank 4 has recently been experiencing RF problems, which may or may not be related to its replaced RF coupler. This situation stimulated a request by Dan Rees to provide EM modeling of the RF drive in the DTL tank 4 (T4). Jim O'Hara provided a CAD model that was imported into the CST Microwave Studio (MWS) and after some modifications became a part of a simplified MWS model of the T4 RF drive. This technical note describes the model and presents simulation results.

  20. DOE-EM-45 PACKAGING OPERATIONS AND MAINTENANCE COURSE

    SciTech Connect

    Watkins, R.; England, J.

    2010-05-28

    Savannah River National Laboratory - Savannah River Packaging Technology (SRNL-SRPT) delivered the inaugural offering of the Packaging Operations and Maintenance Course for DOE-EM-45's Packaging Certification Program (PCP) at the University of South Carolina Aiken on September 1 and 2, 2009. Twenty-nine students registered, attended, and completed this training. The DOE-EM-45 Packaging Certification Program (PCP) sponsored the presentation of a new training course, Packaging Maintenance and Operations, on September 1-2, 2009 at the University of South Carolina Aiken (USC-Aiken) campus in Aiken, SC. The premier offering of the course was developed and presented by the Savannah River National Laboratory, and attended by twenty-nine students across the DOE, NNSA and private industry. This training informed package users of the requirements associated with handling shipping containers at a facility (user) level and provided a basic overview of the requirements typically outlined in Safety Analysis Report for Packaging (SARP) Chapters 1, 7, and 8. The course taught packaging personnel about the regulatory nature of SARPs to help reduce associated and often costly packaging errors. Some of the topics covered were package contents, loading, unloading, storage, torque requirements, maintaining records, how to handle abnormal conditions, lessons learned, leakage testing (including demonstration), and replacement parts. The target audience for this course was facility operations personnel, facility maintenance personnel, and field quality assurance personnel who are directly involved in the handling of shipping containers. The training also aimed at writers of SARP Chapters 1, 7, and 8, package designers, and anyone else involved in radioactive material packaging and transportation safety. Student feedback and critiques of the training were very positive. SRNL will offer the course again at USC Aiken in September 2010.

  1. Development of MPD thruster EM for a space test. [Engineering model

    SciTech Connect

    Shiina, K.; Suzuki, H.; Uematsu, K.; Ohtsuka, T.; Toki, K. Institute of Space and Astronautical Science, Kanagawa )

    1990-01-01

    An engineering model (EM) of MPD thruster has been developed for a space test on board the first Space Flyer Unit (SFU-1). A thermal vacuum test was conducted, and the following results were obtained: (1) a thermal mathematical model of MPD thruster EM was established, (2) sizing data of thruster heaters were obtained, and (3) thermal characteristics of the MPD thruster EM were confirmed to meet the requirement. The data are going to be reflected in designing a protoflight model of MPD thruster. 8 refs.

  2. Analytical laboratory quality assurance guidance in support of EM environmental sampling and analysis activities

    SciTech Connect

    Not Available

    1994-05-01

    This document introduces QA guidance pertaining to design and implementation of laboratory procedures and processes for collecting DOE Environmental Restoration and Waste Management (EM) ESAA (environmental sampling and analysis activities) data. It addresses several goals: identifying key laboratory issues and program elements to EM HQ and field office managers; providing non-prescriptive guidance; and introducing environmental data collection program elements for EM-263 assessment documents and programs. The guidance describes the implementation of laboratory QA elements within a functional QA program (development of the QA program and data quality objectives are not covered here).

  3. EMS Provider Assessment of Vehicle Damage Compared to a Professional Crash Reconstructionist

    PubMed Central

    Lerner, E. Brooke; Cushman, Jeremy T.; Blatt, Alan; Lawrence, Richard; Shah, Manish N.; Swor, Robert; Brasel, Karen; Jurkovich, Gregory J.

    2011-01-01

    Objective To determine the accuracy of EMS provider assessments of motor vehicle damage, when compared to measurements made by a professional crash reconstructionist. Methods EMS providers caring for adult patients injured during a motor vehicle crash and transported to the regional trauma center in a midsized community were interviewed upon ED arrival. The interview collected provider estimates of crash mechanism of injury. For crashes that met a preset severity threshold, the vehicle’s owner was asked to consent to having a crash reconstructionist assess their vehicle. The assessment included measuring intrusion and external auto deformity. Vehicle damage was used to calculate change in velocity. Paired t-test and correlation were used to compare EMS estimates and investigator derived values. Results 91 vehicles were enrolled; of these 58 were inspected and 33 were excluded because the vehicle was not accessible. 6 vehicles had multiple patients. Therefore, a total of 68 EMS estimates were compared to the inspection findings. Patients were 46% male, 28% admitted to hospital, and 1% died. Mean EMS estimated deformity was 18” and mean measured was 14”. Mean EMS estimated intrusion was 5” and mean measured was 4”. EMS providers and the reconstructionist had 67% agreement for determination of external auto deformity (kappa 0.26), and 88% agreement for determination of intrusion (kappa 0.27) when the 1999 Field Triage Decision Scheme Criteria were applied. Mean EMS estimated speed prior to the crash was 48 mph±13 and mean reconstructionist estimated change in velocity was 18 mph±12 (correlation -0.45). EMS determined that 19 vehicles had rolled over while the investigator identified 18 (kappa 0.96). In 55 cases EMS and the investigator agreed on seatbelt use, for the remaining 13 cases there was disagreement (5) or the investigator was unable to make a determination (8) (kappa 0.40). Conclusions This study found that EMS providers are good at estimating

  4. Integration of artificial intelligence applications in the EMS: Issues and solutions

    SciTech Connect

    Bann, J.; Irisarri, G.; Kirschen, D.; Miller, B.; Mokhtari, S.

    1996-02-01

    This paper discusses the issues which must be addressed when integrating Artificial Intelligence (AI) and, in particular, expert system applications in an Energy Management System (EMS) environment. It is argued that these issues can be resolved by creating an environment which supports all the interfaces between the Artificial Intelligence (AI) applications and the EMS. This environment should also be responsible for maintaining a model of the power system common to all the AI applications. Once this environment has been created, AI applications can be easily plugged into the EMS. The design of such an environment is described and case studies of its implementation are provided to illustrate its flexibility.

  5. Integration of artificial intelligence applications in the EMS: Issues and solutions

    SciTech Connect

    Bann, J.; Irisarri, G.; Kirschen, D.; Miller, B.; Mokhtari, S.

    1995-12-31

    This paper discusses the issues which must be addressed when integrating Artificial Intelligence (AI) and, in particular, expert system applications in an Energy Management System (EMS) environment. It is argued that these issues can be resolved by creating an environment which supports all the interfaces between the Artificial Intelligence (AI) applications and the EMS. This environment should also be responsible for maintaining a model of the power system common to all the AI applications. Once this environment has been created, AI applications can be easily plugged into the EMS. The design of such an environment is described and case studies of its implementation are provided to illustrate its flexibility.

  6. 7. PHOTOCOPY, ELEVATIONS FOR E.M. BARRACKS, N.C.O. AND OFFICERS QUARTERS. ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    7. PHOTOCOPY, ELEVATIONS FOR E.M. BARRACKS, N.C.O. AND OFFICERS QUARTERS. - NIKE Missile Base SL-40, Barracks No. 1, North end of base, southest of Basketball Court & northwest of Barracks No. 2, Hecker, Monroe County, IL

  7. Factors that Influence the Formation and Stability of Thin, Cryo-EM Specimens.

    PubMed

    Glaeser, Robert M; Han, Bong-Gyoon; Csencsits, Roseann; Killilea, Alison; Pulk, Arto; Cate, Jamie H D

    2016-02-23

    Poor consistency of the ice thickness from one area of a cryo-electron microscope (cryo-EM) specimen grid to another, from one grid to the next, and from one type of specimen to another, motivates a reconsideration of how to best prepare suitably thin specimens. Here we first review the three related topics of wetting, thinning, and stability against dewetting of aqueous films spread over a hydrophilic substrate. We then suggest that the importance of there being a surfactant monolayer at the air-water interface of thin, cryo-EM specimens has been largely underappreciated. In fact, a surfactant layer (of uncontrolled composition and surface pressure) can hardly be avoided during standard cryo-EM specimen preparation. We thus suggest that better control over the composition and properties of the surfactant layer may result in more reliable production of cryo-EM specimens with the desired thickness.

  8. Sampling quality assurance guidance in support of EM environmental sampling and analysis activities

    SciTech Connect

    Not Available

    1994-05-01

    This document introduces quality assurance guidance pertaining to the design and implementation of sampling procedures and processes for collecting environmental data for DOE`s Office of EM (Environmental Restoration and Waste Management).

  9. Aquisição de Estreptococos Mutans e Desenvolvimento de Cárie Dental em Primogênitos

    PubMed Central

    NOCE, Erica; RUBIRA, Cassia Maria Fischer; da Silva ROSA, Odila Pereira; da SILVA, Salete Moura Bonifácio; BRETZ, Walter Antonio

    2011-01-01

    Objetivo Avaliar o momento de aquisição de estreptococos mutans (EM), desenvolvimento de cárie dental e as variáveis a eles associadas no decorrer de 23 meses, em primogênitos de famílias de baixo nível socioeconômico, desde os sete meses de idade. Método A amostra foi selecionada com base em mães densamente colonizadas por EM, incluindo todos os membros de 14 famílias que conviviam na mesma casa. Foram envolvidos no estudo 14 mães, pais e primogênitos e 8 parentes, na maioria avós. Exames clínicos e radiográficos iniciais determinaram os índices de cárie e condição periodontal dos adultos. Contagens de EM foram feitas em todos os adultos nas duas primeiras visitas. Nas crianças foram avaliados os níveis de EM, o número de dentes e de cáries, em quatro visitas. Resultados A prevalência de EM nos adultos foi alta, estando ausente em apenas um dos pais. EM foram detectados em 1, 2, 3 e 10 crianças, respectivamente nas visitas #1, 2, 3 e 4. A cárie dental foi detectada em apenas três crianças na última visita (aos 30 meses de idade), as quais apresentaram escores de EM significantemente maiores que as crianças sem cárie, na mesma visita. Conclusão Exclusivamente a condição social de baixa renda e mães densamente colonizadas por EM não são sinônimo de colonização precoce e alta atividade de cárie em crianças cuidadas em casa. O desenvolvimento de cárie está significantemente associado a escores elevados de EM nas crianças. PMID:22022218

  10. The Soils and Groundwater – EM-20 S&T Roadmap Quality Assurance Project Plan

    SciTech Connect

    Fix, N. J.

    2008-02-11

    The Soils and Groundwater – EM-20 Science and Technology Roadmap Project is a U.S. Department of Energy, Office of Environmental Management-funded initiative designed to develop new methods, strategies and technology for characterizing, modeling, remediating, and monitoring soils and groundwater contaminated with metals, radionuclides, and chlorinated organics. This Quality Assurance Project Plan provides the quality assurance requirements and processes that will be followed by EM-20 Roadmap Project staff.

  11. Effects of Loaded Squat Exercise with and without Application of Superimposed EMS on Physical Performance

    PubMed Central

    Wirtz, Nicolas; Zinner, Christoph; Doermann, Ulrike; Kleinoeder, Heinz; Mester, Joachim

    2016-01-01

    The aim of the present study was to investigate the effects of a multiple set squat exercise training intervention with superimposed electromyostimulation (EMS) on strength and power, sprint and jump performance. Twenty athletes from different disciplines participated and were divided into two groups: strength training (S) or strength training with superimposed EMS (S+E). Both groups completed the same training program twice a week over a six week period consisting of four sets of the 10 repetition maximum of back squats. Additionally, the S+E group had EMS superimposed to the squat exercise with simultaneous stimulation of leg and trunk muscles. EMS intensity was adjusted to 70% of individual pain threshold to ensure dynamic movement. Strength and power of different muscle groups, sprint, and vertical jump performance were assessed one week before (pre), one week after (post) and three weeks (re) following the training period. Both groups showed improvements in leg press strength and power, countermovement and squat jump performance and pendulum sprint (p < 0.05), with no changes for linear sprint. Differences between groups were only evident at the leg curl machine with greater improvements for the S+E group (p < 0.05). Common squat exercise training and squat exercise with superimposed EMS improves maximum strength and power, as well as jumping abilities in athletes from different disciplines. The greater improvements in strength performance of leg curl muscles caused by superimposed EMS with improvements in strength of antagonistic hamstrings in the S+E group are suggesting the potential of EMS to unloaded (antagonistic) muscle groups. Key points Similar strength adaptations occurred after a 6 week 10 RM back squat exercise program with superimposed EMS (S+E) and 10 RM back squat exercise (S) alone. Specific adaptations for S+E at the leg curl muscles were evident. S and S+E improved SJ, CMJ and pendulum sprint performance. No improvement occurred in linear

  12. Estudo comparativo entre estrelas centrais de nebulosas planetárias deficientes em hidrogênio

    NASA Astrophysics Data System (ADS)

    Marcolino, W. L. F.; de Araújo, F. X.

    2003-08-01

    Apresentamos neste trabalho o resultado de um estudo das principais características espectrais das estrelas centrais de nebulosas planetárias (ECNP) deficientes em hidrogênio. A origem e a evolução dessas estrelas ainda constitui um problema em aberto na evolução estelar. Geralmente esses objetos são divididos em [WCE], [WCL] e [WELS]. Os tipos [WCE] e [WCL] apresentam um espectro típico de uma estrela Wolf-Rayet carbonada de população I e as [WELS] apresentam linhas fracas de carbono e oxigênio em emissão. Existem evidências que apontam a seguinte sequência evolutiva : [WCL] = > [WCE] = > [WELS] = > PG 1159 (pré anã-branca). No entanto, tal cenário apresenta falhas como por exemplo a falta de ECNP entre os tipos [WCL] e [WCE]. Baseados em uma amostra de 24 objetos obtida no telescópio de 1.52m em La Silla, Chile (acordo ESO/ON), ao longo do ano 2000, apresentamos os resultados da comparação das larguras equivalentes de diversas linhas relevantes entre os tipos [WCL], [WCE] e [WELS]. Verificamos que nossos dados estão de acordo com a sequência evolutiva. Baseado nas linhas de C IV, conseguimos dividir pela primeira vez as [WELS] em dois grupos principais. Além disso, os dados reforçam a afirmação de que as [WCE] são as estrelas que possuem a maior temperatura entre as ECNP deficientes em hidrogênio. Discutimos ainda, a escassez de dados disponíveis na literatura e a necessidade da obtenção de parametros físicos para estes objetos.

  13. Sobre o uso das séries de Puiseux em mecanica celeste

    NASA Astrophysics Data System (ADS)

    Miloni, O. I.

    2003-08-01

    Neste trabalho é apresentada uma demonstração do uso dos diferentes desenvolvimentos em séries para as equações de perturbação em Mecânica Celeste no marco Hamiltoniano. Em trabalhos clássicos como os de Poincaré (Poincaré, 1893) por exemplo, já esta planteado o uso de potências não inteiras no pequeno parâmetro, o que evidencia a não analiticidade das funções quando uma ressonância ocorre. Nestes trabalhos os desenvolvimentos são na raíz quadrada da massa de Júpiter (o pequeno parâmetro). Mais recentemente (Ferraz-Mello, 1985) outros tipos de desenvolvimentos foram aplicados modificando substancialmente as ordens de grandeza e a velocidade de convergência das séries. Com esta abordagem, os desenvolvimentos foram expressados em termos da raíz cúbica do pequeno parâmetro. Neste trabalho apresentamos um enfoque geral, onde os diferentes tipos de desenvolvimentos em séries de Puiseux (Valiron, 1950) são obtidos a partir da aplicação de Teorema de Preparação de Weierstrass (Goursat, 1916) considerando a equação de Hamilton-Jacobi como uma equação algébrica. Os resultados são aplicados ao problema restrito dos três corpos em ressonância de primeira ordem e, dependendo da grandeza da excentricidade do asteróide em relação à de Júpiter, obtemos os diferentes desenvolvimentos, em raíz quadrada ou raíz cúbica da massa de Júpiter.

  14. Self-assembled monolayers improve protein distribution on holey carbon cryo-EM supports

    PubMed Central

    Meyerson, Joel R.; Rao, Prashant; Kumar, Janesh; Chittori, Sagar; Banerjee, Soojay; Pierson, Jason; Mayer, Mark L.; Subramaniam, Sriram

    2014-01-01

    Poor partitioning of macromolecules into the holes of holey carbon support grids frequently limits structural determination by single particle cryo-electron microscopy (cryo-EM). Here, we present a method to deposit, on gold-coated carbon grids, a self-assembled monolayer whose surface properties can be controlled by chemical modification. We demonstrate the utility of this approach to drive partitioning of ionotropic glutamate receptors into the holes, thereby enabling 3D structural analysis using cryo-EM methods. PMID:25403871

  15. EMS Provider Attitudes and Perceptions of Enrolling Patients without Consent in Prehospital Emergency Research.

    PubMed

    Jasti, Jamie; Fernandez, Antonio R; Schmidt, Terri A; Lerner, E Brooke

    2016-01-01

    The purpose of this study was to evaluate the attitudes and opinions of a broad population of EMS providers on enrolling patients in research without consent. A survey was conducted in 2010 of all EMS providers who participated in the National Registry of Emergency Medical Technicians (NREMT) reregistration process, which included half of all registered providers. Each reregistration packet included our optional survey, which had nine 6-point Likert scale questions concerning their opinion of research studies without consent as well as 8 demographic questions. Responses were collapsed to agree and disagree and then analyzed using descriptive statistics with 99% confidence intervals. A total of 65,993 EMS providers received the survey and 23,832 (36%) participated. Most respondents agreed (98.4%, 99%CI: 98.2-98.6) that EMS research is important, but only 30.9% (99%CI: 30.1-31.6) agreed with enrolling patients without their consent when it is important to learn about a new treatment. Only 46.6% (99%Cl: 45.7-47.4) were personally willing to be enrolled in a study without their consent. A majority (68.5% [99%Cl: 67.7-69.3]) of respondents believed that EMS providers should have the individual right to refuse to enroll patients in EMS research. While the majority of respondents agreed that EMS research is important, considerably less agree with enrolling patients without consent and less than half would be willing to be enrolled in a study without their consent. Prior to starting an Exception from Informed Consent (EFIC) study, researchers should discuss with EMS providers their perceptions of enrolling patients without consent and address their concerns.

  16. The development of cryo-EM into a mainstream structural biology technique

    PubMed Central

    Nogales, Eva

    2016-01-01

    Single-particle cryo-electron microscopy (cryo-EM) has emerged over the last two decades as a technique capable of studying challenging systems that otherwise defy structural characterization. Recent technical advances have resulted in a ‘quantum leap’ in applicability, throughput and achievable resolution that has gained this technique worldwide attention. Here I discuss some of the major historical landmarks in the development of the cryo-EM field, ultimately leading to its present success. PMID:27110629

  17. Pre-flight risk assessment in emergency medical service (EMS) helicopters

    NASA Technical Reports Server (NTRS)

    Shively, Robert J.

    1990-01-01

    A preflight risk assessment system (SAFE) was developed at NASA-Ames Research Center for civil EMS operations to assist pilots in making a decision objectively to accept or decline a mission. The ability of the SAFE system to predict risk profiles was examined at an EMS operator. Results of this field study showed that the usefulness of SAFE was largely dependent on the type of mission flown.

  18. Single-particle cryo-EM data acquisition by using direct electron detection camera.

    PubMed

    Wu, Shenping; Armache, Jean-Paul; Cheng, Yifan

    2016-02-01

    Recent advances in single-particle electron cryo-microscopy (cryo-EM) were largely facilitated by the application of direct electron detection cameras. These cameras feature not only a significant improvement in detective quantum efficiency but also a high frame rate that enables images to be acquired as 'movies' made of stacks of many frames. In this review, we discuss how the applications of direct electron detection cameras in cryo-EM have changed the way the data are acquired.

  19. Effects of Loaded Squat Exercise with and without Application of Superimposed EMS on Physical Performance.

    PubMed

    Wirtz, Nicolas; Zinner, Christoph; Doermann, Ulrike; Kleinoeder, Heinz; Mester, Joachim

    2016-03-01

    The aim of the present study was to investigate the effects of a multiple set squat exercise training intervention with superimposed electromyostimulation (EMS) on strength and power, sprint and jump performance. Twenty athletes from different disciplines participated and were divided into two groups: strength training (S) or strength training with superimposed EMS (S+E). Both groups completed the same training program twice a week over a six week period consisting of four sets of the 10 repetition maximum of back squats. Additionally, the S+E group had EMS superimposed to the squat exercise with simultaneous stimulation of leg and trunk muscles. EMS intensity was adjusted to 70% of individual pain threshold to ensure dynamic movement. Strength and power of different muscle groups, sprint, and vertical jump performance were assessed one week before (pre), one week after (post) and three weeks (re) following the training period. Both groups showed improvements in leg press strength and power, countermovement and squat jump performance and pendulum sprint (p < 0.05), with no changes for linear sprint. Differences between groups were only evident at the leg curl machine with greater improvements for the S+E group (p < 0.05). Common squat exercise training and squat exercise with superimposed EMS improves maximum strength and power, as well as jumping abilities in athletes from different disciplines. The greater improvements in strength performance of leg curl muscles caused by superimposed EMS with improvements in strength of antagonistic hamstrings in the S+E group are suggesting the potential of EMS to unloaded (antagonistic) muscle groups. Key pointsSimilar strength adaptations occurred after a 6 week 10 RM back squat exercise program with superimposed EMS (S+E) and 10 RM back squat exercise (S) alone.Specific adaptations for S+E at the leg curl muscles were evident.S and S+E improved SJ, CMJ and pendulum sprint performance.No improvement occurred in linear sprint

  20. ModEM: A modular system for inversion of elecgtromagnetic geophysical data

    NASA Astrophysics Data System (ADS)

    Egbert, G. D.; Kelbert, A.; Meqbel, N.; Weng, A.

    2010-12-01

    We have developed a modular system of computer codes for inversion of electromagnetic (EM) geophysical data (ModEM). ModEM allows for rapid adaptation of inversion algorithms developed for one purpose (e.g., three-dimensional magnetotellurics (MT)) to other EM problems (e.g., controlled source EM). The modular approach can also simplify maintenance of the inversion code, as well as development of new capabilities -- e.g., allowing for new data types such as the inter-site transfer functions in MT, or modifying model regularization. Basic data objects (model parameters, solution vectors, data vectors) are treated as abstract data types, with a standard set of methods developed for each class, including creation and destruction, and, as appropriate, linear algebra or other vector space methods. Operators required for gradient computations are developed as mappings between these basic object classes. Only these abstract data objects and mappings are manipulated by higher level Jacobian and inversion routines, with no reference to the problem specific details required for a specific EM method, or for the numerical implementation of the forward solver. Required problem-specific components are instantiated at the lowest levels of the system, with details hidden from generic top-level routines by an interface layer. Parallelization using MPI has been implemented at the top level, and is thus applicable to any problem embedded in ModEM. To illustrate the flexibility of the system, we consider applications to two- and three-dimensional MT inversion, as well as simple controlled source EM problems.

  1. Fusion to a homo-oligomeric scaffold allows cryo-EM analysis of a small protein.

    PubMed

    Coscia, Francesca; Estrozi, Leandro F; Hans, Fabienne; Malet, Hélène; Noirclerc-Savoye, Marjolaine; Schoehn, Guy; Petosa, Carlo

    2016-01-01

    Recent technical advances have revolutionized the field of cryo-electron microscopy (cryo-EM). However, most monomeric proteins remain too small (<100 kDa) for cryo-EM analysis. To overcome this limitation, we explored a strategy whereby a monomeric target protein is genetically fused to a homo-oligomeric scaffold protein and the junction optimized to allow the target to adopt the scaffold symmetry, thereby generating a chimeric particle suitable for cryo-EM. To demonstrate the concept, we fused maltose-binding protein (MBP), a 40 kDa monomer, to glutamine synthetase, a dodecamer formed by two hexameric rings. Chimeric constructs with different junction lengths were screened by biophysical analysis and negative-stain EM. The optimal construct yielded a cryo-EM reconstruction that revealed the MBP structure at sub-nanometre resolution. These findings illustrate the feasibility of using homo-oligomeric scaffolds to enable cryo-EM analysis of monomeric proteins, paving the way for applying this strategy to challenging structures resistant to crystallographic and NMR analysis. PMID:27485862

  2. Operation Protective Edge - A Unique Challenge for a Civilian EMS Agency.

    PubMed

    Jaffe, Eli; Strugo, Refael; Wacht, Oren

    2015-10-01

    During July through August 2014, Operation Protective Edge, a military conflict between Israel and the Hamas regime in Gaza, dramatically affected both populations. Magen David Adom (MDA), the Israeli national Emergency Medical Service (EMS) and a member of the Red Cross, faced a unique challenge during the conflict: to continue providing crucial service to the entire civilian population of Israel, which was under constant missile threat. This challenge included not only providing immediate care for routine EMS calls under missile threat, but also preparing and delivering immediate care to civilians injured in attacks on major cities, as well as small communities, in Israel. This task is a challenge for a civilian EMS agency that normally operates in a non-military environment, yet, in an instant, must enhance its capability to respond to a considerable threat to its population. During Operation Protective Edge, MDA provided care for 842 wounded civilians and utilized a significant amount of its resources. Providing EMS services for a civilian population in a mixed civilian/military scenario is a challenging task on a national level for an EMS system, especially when the threat lasts for weeks. This report describes MDA's preparedness and operations during Operation Protective Edge, and the unique EMS challenges and dilemmas the agency faced.

  3. Visualizing Proteins and Macromolecular Complexes by Negative Stain EM: from Grid Preparation to Image Acquisition

    PubMed Central

    Booth, David S.; Avila-Sakar, Agustin; Cheng, Yifan

    2011-01-01

    Single particle electron microscopy (EM), of both negative stained or frozen hydrated biological samples, has become a versatile tool in structural biology 1. In recent years, this method has achieved great success in studying structures of proteins and macromolecular complexes 2, 3. Compared with electron cryomicroscopy (cryoEM), in which frozen hydrated protein samples are embedded in a thin layer of vitreous ice 4, negative staining is a simpler sample preparation method in which protein samples are embedded in a thin layer of dried heavy metal salt to increase specimen contrast 5. The enhanced contrast of negative stain EM allows examination of relatively small biological samples. In addition to determining three-dimensional (3D) structure of purified proteins or protein complexes 6, this method can be used for much broader purposes. For example, negative stain EM can be easily used to visualize purified protein samples, obtaining information such as homogeneity/heterogeneity of the sample, formation of protein complexes or large assemblies, or simply to evaluate the quality of a protein preparation. In this video article, we present a complete protocol for using an EM to observe negatively stained protein sample, from preparing carbon coated grids for negative stain EM to acquiring images of negatively stained sample in an electron microscope operated at 120kV accelerating voltage. These protocols have been used in our laboratory routinely and can be easily followed by novice users. PMID:22215030

  4. An Uncertainty Analysis for Predicting Soil Profile Salinity Using EM Induction Data

    NASA Astrophysics Data System (ADS)

    Huang, Jingyi; Monteiro Santos, Fernando; Triantafilis, John

    2016-04-01

    Proximal soil sensing techniques such as electromagnetic (EM) induction have been used to identify and map the areal variation of average soil properties. However, soil varies with depth owing to the action of various soil forming factors (e.g., parent material and topography). In this work we collected EM data using an EM38 and EM34 meter along a 22-km transect in the Trangie District, Australia.We jointly inverted these data using EM4Soil software and compare our 2-dimensional model of true electrical conductivity (sigma - mS/m) with depth against measured electrical conductivity of a saturated soil-paste extract (ECe - dS/m) at depth of 0-16 m. Through the use of a linear regression (LR) model and by varying forward modelling algorithms (cumulative function and full solution), inversion algorithms (S1 and S2), and damping factor (lambda) we determined a suitable electromagnetic conductivity image (EMCI) which was optimal when using the full solution, S2 and lambda = 0.6. To evaluate uncertainty of the inversion process and the LR model, we conducted an uncertainty analysis. The distribution of the model misfit shows the largest uncertainty caused by inversion (mostly due to EM34-40) occurs at deeper profiles while the largest uncertainty of the LR model occurs where the soil profile is most saline. These uncertainty maps also illustrate us how the model accuracy can be improved in the future.

  5. Fusion to a homo-oligomeric scaffold allows cryo-EM analysis of a small protein.

    PubMed

    Coscia, Francesca; Estrozi, Leandro F; Hans, Fabienne; Malet, Hélène; Noirclerc-Savoye, Marjolaine; Schoehn, Guy; Petosa, Carlo

    2016-08-03

    Recent technical advances have revolutionized the field of cryo-electron microscopy (cryo-EM). However, most monomeric proteins remain too small (<100 kDa) for cryo-EM analysis. To overcome this limitation, we explored a strategy whereby a monomeric target protein is genetically fused to a homo-oligomeric scaffold protein and the junction optimized to allow the target to adopt the scaffold symmetry, thereby generating a chimeric particle suitable for cryo-EM. To demonstrate the concept, we fused maltose-binding protein (MBP), a 40 kDa monomer, to glutamine synthetase, a dodecamer formed by two hexameric rings. Chimeric constructs with different junction lengths were screened by biophysical analysis and negative-stain EM. The optimal construct yielded a cryo-EM reconstruction that revealed the MBP structure at sub-nanometre resolution. These findings illustrate the feasibility of using homo-oligomeric scaffolds to enable cryo-EM analysis of monomeric proteins, paving the way for applying this strategy to challenging structures resistant to crystallographic and NMR analysis.

  6. Using Molecular Simulation to Model High-Resolution Cryo-EM Reconstructions.

    PubMed

    Kirmizialtin, Serdal; Loerke, Justus; Behrmann, Elmar; Spahn, Christian M T; Sanbonmatsu, Karissa Y

    2015-01-01

    An explosion of new data from high-resolution cryo-electron microscopy (cryo-EM) studies has produced a large number of data sets for many species of ribosomes in various functional states over the past few years. While many methods exist to produce structural models for lower resolution cryo-EM reconstructions, high-resolution reconstructions are often modeled using crystallographic techniques and extensive manual intervention. Here, we present an automated fitting technique for high-resolution cryo-EM data sets that produces all-atom models highly consistent with the EM density. Using a molecular dynamics approach, atomic positions are optimized with a potential that includes the cross-correlation coefficient between the structural model and the cryo-EM electron density, as well as a biasing potential preserving the stereochemistry and secondary structure of the biomolecule. Specifically, we use a hybrid structure-based/ab initio molecular dynamics potential to extend molecular dynamics fitting. In addition, we find that simulated annealing integration, as opposed to straightforward molecular dynamics integration, significantly improves performance. We obtain atomistic models of the human ribosome consistent with high-resolution cryo-EM reconstructions of the human ribosome. Automated methods such as these have the potential to produce atomistic models for a large number of ribosome complexes simultaneously that can be subsequently refined manually.

  7. Cryo-EM Data Are Superior to Contact and Interface Information in Integrative Modeling.

    PubMed

    de Vries, Sjoerd J; Chauvot de Beauchêne, Isaure; Schindler, Christina E M; Zacharias, Martin

    2016-02-23

    Protein-protein interactions carry out a large variety of essential cellular processes. Cryo-electron microscopy (cryo-EM) is a powerful technique for the modeling of protein-protein interactions at a wide range of resolutions, and recent developments have caused a revolution in the field. At low resolution, cryo-EM maps can drive integrative modeling of the interaction, assembling existing structures into the map. Other experimental techniques can provide information on the interface or on the contacts between the monomers in the complex. This inevitably raises the question regarding which type of data is best suited to drive integrative modeling approaches. Systematic comparison of the prediction accuracy and specificity of the different integrative modeling paradigms is unavailable to date. Here, we compare EM-driven, interface-driven, and contact-driven integrative modeling paradigms. Models were generated for the protein docking benchmark using the ATTRACT docking engine and evaluated using the CAPRI two-star criterion. At 20 Å resolution, EM-driven modeling achieved a success rate of 100%, outperforming the other paradigms even with perfect interface and contact information. Therefore, even very low resolution cryo-EM data is superior in predicting heterodimeric and heterotrimeric protein assemblies. Our study demonstrates that a force field is not necessary, cryo-EM data alone is sufficient to accurately guide the monomers into place. The resulting rigid models successfully identify regions of conformational change, opening up perspectives for targeted flexible remodeling.

  8. New Developments in the Technology Readiness Assessment Process in US DOE-EM - 13247

    SciTech Connect

    Krahn, Steven; Sutter, Herbert; Johnson, Hoyt

    2013-07-01

    A Technology Readiness Assessment (TRA) is a systematic, metric-based process and accompanying report that evaluates the maturity of the technologies used in systems; it is designed to measure technology maturity using the Technology Readiness Level (TRL) scale pioneered by the National Aeronautics and Space Administration (NASA) in the 1980's. More recently, DoD has adopted and provided systematic guidance for performing TRAs and determining TRLs. In 2007 the GAO recommended that the DOE adopt the NASA/DoD methodology for evaluating technology maturity. Earlier, in 2006-2007, DOE-EM had conducted pilot TRAs on a number of projects at Hanford and Savannah River. In March 2008, DOE-EM issued a process guide, which established TRAs as an integral part of DOE-EM's Project Management Critical Decision Process. Since the development of its detailed TRA guidance in 2008, DOE-EM has continued to accumulate experience in the conduct of TRAs and the process for evaluating technology maturity. DOE has developed guidance on TRAs applicable department-wide. DOE-EM's experience with the TRA process, the evaluations that led to recently developed proposed revisions to the DOE-EM TRA/TMP Guide; the content of the proposed changes that incorporate the above lessons learned and insights are described. (authors)

  9. An erythromycin derivative, EM-523, induces motilin-like gastrointestinal motility in dogs.

    PubMed

    Inatomi, N; Satoh, H; Maki, Y; Hashimoto, N; Itoh, Z; Omura, S

    1989-11-01

    The effect of an erythromycin derivative, EM-523, on gastrointestinal motility was investigated in conscious dogs and compared with that of motilin cisapride, trimebutine and metoclopramide. In the fasting state, EM-523 given i.v. or i.d. at 3 micrograms/kg or more induced contractions in the stomach that migrated along the small intestine. The pattern of the contractions was very similar to that induced by motilin. In the digestive state, EM-523 increased the amplitude of gastric contractions. Cisapride and metoclopramide increased gastrointestinal motility both in the fasting and digestive states; however, their contractile pattern was different from that of EM-523. Trimebutine did not induce gastric motility in the fasting state but rather decreased gastric motility in the digestive state. The contractions induced by EM-523 and motilin were inhibited by atropine but were not affected by naloxone, suggesting that the cholinergic pathway is important in the exertion of their action. These results indicate that EM-523 mimics motilin in stimulating gastrointestinal motility and that this agent may be useful treat gastrointestinal disorders such as gastric stasis, gastroesophageal reflux, and postoperative ileus, and so forth. PMID:2810120

  10. Fusion to a homo-oligomeric scaffold allows cryo-EM analysis of a small protein

    PubMed Central

    Coscia, Francesca; Estrozi, Leandro F.; Hans, Fabienne; Malet, Hélène; Noirclerc-Savoye, Marjolaine; Schoehn, Guy; Petosa, Carlo

    2016-01-01

    Recent technical advances have revolutionized the field of cryo-electron microscopy (cryo-EM). However, most monomeric proteins remain too small (<100 kDa) for cryo-EM analysis. To overcome this limitation, we explored a strategy whereby a monomeric target protein is genetically fused to a homo-oligomeric scaffold protein and the junction optimized to allow the target to adopt the scaffold symmetry, thereby generating a chimeric particle suitable for cryo-EM. To demonstrate the concept, we fused maltose-binding protein (MBP), a 40 kDa monomer, to glutamine synthetase, a dodecamer formed by two hexameric rings. Chimeric constructs with different junction lengths were screened by biophysical analysis and negative-stain EM. The optimal construct yielded a cryo-EM reconstruction that revealed the MBP structure at sub-nanometre resolution. These findings illustrate the feasibility of using homo-oligomeric scaffolds to enable cryo-EM analysis of monomeric proteins, paving the way for applying this strategy to challenging structures resistant to crystallographic and NMR analysis. PMID:27485862

  11. EM techniques for archaeological laboratory experiments: preliminary results

    NASA Astrophysics Data System (ADS)

    Capozzoli, Luigi; De Martino, Gregory; Giampaolo, Valeria; Raffaele, Luongo; Perciante, Felice; Rizzo, Enzo

    2015-04-01

    The electromagnetic techniques (EM) are based on the investigation of subsoil geophysical parameters and in the archaeological framework they involve in studying contrasts between the buried cultural structures and the surrounding materials. Unfortunately, the geophysical contrast between archaeological features and surrounding soils sometimes are difficult to define due to problems of sensitivity and resolution both related on the characteristic of the subsoil and the geophysical methods. For this reason an experimental activity has been performed in the Hydrogeosite laboratory addressed on the assessment of the capability of geophysical techniques to detect archeological remains placed in the humid/saturated subsoil. At Hydrogeosite Laboratory of CNR-IMAA, a large scale sand-box is located, consisting on a pool shape structures of 230m3 where archaeological remains have been installed . The remains are relative to a living environment and burial of Roman times (walls, tombs, roads, harbour, etc.) covered by sediments. In order to simulate lacustrine and wetland condition and to simulate extreme events (for example underwater landslide, fast natural erosion coast, etc.) the phreatic level was varied and various acquisitions for the different scenarios were performed. In order to analyze the EM behavior of the buried small archaeological framework, ground penetrating radar (GPR) and electrical resistivity tomographies were performed. With GPR, analysis in time domain and frequency domain were performed and coupled to information obtained through resistivity analysis with the support of numerical simulations used to compare the real data with those modeled. A dense grid was adopted for 400 and 900 MHz e-m acquisitions in both the directions, the maximum depth of investigation was limited and less than 3 meters. The same approach was used for ERT acquisition where different array are employed, in particular 3D configuration was used to carry out a 3D resistivity

  12. Abundâncias em estrelas de Bário

    NASA Astrophysics Data System (ADS)

    Allen, D. M.

    2003-08-01

    Estrelas de Bário apresentam linhas intensas de elementos produzidos pelo processos (ex: Ba, Y, Sr, Zr) e bandas intensas de CN, C2 e CH. A hipótese mais aceita sobre a origem deste grupo peculiar é a de que essas estrelas façam parte de sistemas binários, tendo recebido material enriquecido em elementos pesados da companheira mais evoluída. Apresentamos neste trabalho uma análise detalhada de uma amostra de estrelas desta classe, incluindo determinação de parâmetros atmosféricos e cálculo de abundâncias. As temperaturas efetivas foram determinadas a partir de dados fotométricos obtidos com o Fotrap instalado no telescópio Zeiss do LNA (Laboratório Nacional de Astrofísica) (B-V, V-I, R-I, V-R), e coletados na literatura nos catálogos Hipparcos (B-V), 2MASS (Two Micron All Sky Survey) (V-K) e The General Catalogue Photometric Data (sistema Geneva). Obtivemos uma faixa de temperaturas de 4400 £ Tef £ 6500. As metalicidades foram determinadas a partir de linhas de Fe I e Fe II, estando os resultados no intervalo -1 £ [Fe/H] £ +0.1. O log g foi determinado pelo equilíbrio de ionização e pela relação com a magnitude bolométrica, a temperatura e a massa, sendo os resultados na faixa 1.5 £ log g £ 4.5. As distâncias utilizadas foram determinadas com o auxílio das paralaxes Hipparcos, e as massas determinadas por modelos de isócronas. Os espectros utilizados foram obtidos com o espectrógrafo FEROS no Telescópio de 1,5m do ESO (European Southern Observatory). As abundâncias foram calculadas por meio de síntese espectral de linhas individuais incluindo elementos alfa, pico do Fe, s e r. Encontramos um excesso de elementos pesados em relação ao Fe, como esperado para estrelas de Bário.

  13. Tide-driven fluid mud transport in the Ems estuary

    NASA Astrophysics Data System (ADS)

    Becker, Marius; Maushake, Christian; Winter, Christian

    2014-05-01

    The Ems estuary, located at the border between The Netherlands and Germany, experienced a significant change of the hydrodynamic regime during the past decades, as a result of extensive river engineering. With the net sediment transport now being flood-oriented, suspended sediment concentrations have increased dramatically, inducing siltation and formation of fluid mud layers, which, in turn, influence hydraulic flow properties, such as turbulence and the apparent bed roughness. Here, the process-based understanding of fluid mud is essential to model and predict mud accumulation, not only regarding the anthropogenic impact, but also in view of the expected changes of environmental boundary conditions, i.e., sea level rise. In the recent past, substantial progress has been made concerning the understanding of estuarine circulation and influence of tidal asymmetry on upstream sediment accumulation. While associated sediment transport formulations have been implemented in the framework of numerical modelling systems, in-situ data of fluid mud are scarce. This study presents results on tide-driven fluid mud dynamics, measured during four tidal cycles aside the navigation channel in the Ems estuary. Lutoclines, i.e., strong vertical density gradients, were detected by sediment echo sounder (SES). Acoustic Doppler current profiles (ADCP) of different acoustic frequencies were used to determine hydrodynamic parameters and the vertical distribution of suspended sediment concentrations in the upper part of the water column. These continuous profiling measurements were complemented by CTD, ADV, and OBS casts. SES and ADCP profiles show cycles of fluid mud entrainment during accelerating flow, and subsequent settling, and the reformation of a lutocline during decelerating flow and slack water. Significant differences are revealed between flood and ebb phase. Highest entrainment rates are measured at the beginning of the flood phase, associated with strong current shear and

  14. Cryo-EM Structure Determination Using Segmented Helical Image Reconstruction.

    PubMed

    Fromm, S A; Sachse, C

    2016-01-01

    Treating helices as single-particle-like segments followed by helical image reconstruction has become the method of choice for high-resolution structure determination of well-ordered helical viruses as well as flexible filaments. In this review, we will illustrate how the combination of latest hardware developments with optimized image processing routines have led to a series of near-atomic resolution structures of helical assemblies. Originally, the treatment of helices as a sequence of segments followed by Fourier-Bessel reconstruction revealed the potential to determine near-atomic resolution structures from helical specimens. In the meantime, real-space image processing of helices in a stack of single particles was developed and enabled the structure determination of specimens that resisted classical Fourier helical reconstruction and also facilitated high-resolution structure determination. Despite the progress in real-space analysis, the combination of Fourier and real-space processing is still commonly used to better estimate the symmetry parameters as the imposition of the correct helical symmetry is essential for high-resolution structure determination. Recent hardware advancement by the introduction of direct electron detectors has significantly enhanced the image quality and together with improved image processing procedures has made segmented helical reconstruction a very productive cryo-EM structure determination method.

  15. TRLFS: Analysing spectra with an expectation-maximization (EM) algorithm

    NASA Astrophysics Data System (ADS)

    Steinborn, A.; Taut, S.; Brendler, V.; Geipel, G.; Flach, B.

    2008-12-01

    A new approach for fitting statistical models to time-resolved laser-induced fluorescence spectroscopy (TRLFS) spectra is presented. Such spectra result from counting emitted photons in defined intervals. Any photon can be described by emission time and wavelength as observable attributes and by component and peak affiliation as hidden ones. Understanding the attribute values of the emitted photons as drawn from a probability density distribution, the model estimation problem can be described as a statistical problem with incomplete data. To solve the maximum likelihood task, an expectation-maximization (EM) algorithm is derived and tested. In contrast to the well known least squares method, the advantage of the new approach is its ability to decompose the spectrum into its components and peaks using the revealed hidden attributes of the photons as well as the ability to decompose a background-superimposed spectrum into the exploitable signal of the fluorescent chemical species and the background. This facilitates new possibilities for evaluation of the resulting model parameters. The simultaneous detection of temporal and spectral model parameters provides a mutually consistent description of TRLFS spectra.

  16. Primary Mechanism of EM Interaction with the Live Tissues

    NASA Astrophysics Data System (ADS)

    Musakhanyan, Viktor

    There is a prevailing opinion that the theoretical explanation of electromagnetic (EM) fields influence on live organisms is impossible to explain theoretically and even the play between parameters of waves and tissues is unknown to us. The explanation of mechanism of this influence is vitally important owing to the development of new types of electronic devices operating in different frequency ranges and due to the still continuing controversy about their adverse health effect. It is shown that the application of newly developed procedure of shutting-on of the interaction of charged particles with electromagnetic fields allows explaining their influence on live tissue by origination of macroscopic polarization currents due to the joint action of electric and magnetic components of electromagnetic waves. The currents originate in the case of resonance between the proper frequency of the medium and of frequency of external electromagnetic fields. Thus, the experiments to measure these polarization currents can provide information about dangerous frequency ranges and these ranges, with maximal polarization currents, should be excluded during construction of electronic devices.

  17. PREFACE: EmQM13: Emergent Quantum Mechanics 2013

    NASA Astrophysics Data System (ADS)

    2014-04-01

    These proceedings comprise the invited lectures of the second international symposium on Emergent Quantum Mechanics (EmQM13), which was held at the premises of the Austrian Academy of Sciences in Vienna, Austria, 3-6 October 2013. The symposium was held at the ''Theatersaal'' of the Academy of Sciences, and was devoted to the open exploration of emergent quantum mechanics, a possible ''deeper level theory'' that interconnects three fields of knowledge: emergence, the quantum, and information. Could there appear a revised image of physical reality from recognizing new links between emergence, the quantum, and information? Could a novel synthesis pave the way towards a 21st century, ''superclassical'' physics? The symposium provided a forum for discussing (i) important obstacles which need to be overcome as well as (ii) promising developments and research opportunities on the way towards emergent quantum mechanics. Contributions were invited that presented current advances in both standard as well as unconventional approaches to quantum mechanics. The EmQM13 symposium was co-organized by Gerhard Grössing (Austrian Institute for Nonlinear Studies (AINS), Vienna), and by Jan Walleczek (Fetzer Franklin Fund, USA, and Phenoscience Laboratories, Berlin). After a very successful first conference on the same topic in 2011, the new partnership between AINS and the Fetzer Franklin Fund in producing the EmQM13 symposium was able to further expand interest in the promise of emergent quantum mechanics. The symposium consisted of two parts, an opening evening addressing the general public, and the scientific program of the conference proper. The opening evening took place at the Great Ceremonial Hall (Grosser Festsaal) of the Austrian Academy of Sciences, and it presented talks and a panel discussion on ''The Future of Quantum Mechanics'' with three distinguished speakers: Stephen Adler (Princeton), Gerard 't Hooft (Utrecht) and Masanao Ozawa (Nagoya). The articles contained in

  18. Specimen Preparation for High-Resolution Cryo-EM.

    PubMed

    Passmore, L A; Russo, C J

    2016-01-01

    Imaging a material with electrons at near-atomic resolution requires a thin specimen that is stable in the vacuum of the transmission electron microscope. For biological samples, this comprises a thin layer of frozen aqueous solution containing the biomolecular complex of interest. The process of preparing a high-quality specimen is often the limiting step in the determination of structures by single-particle electron cryomicroscopy (cryo-EM). Here, we describe a systematic approach for going from a purified biomolecular complex in aqueous solution to high-resolution electron micrographs that are suitable for 3D structure determination. This includes a series of protocols for the preparation of vitrified specimens on various supports, including all-gold and graphene. We also describe techniques for troubleshooting when a preparation fails to yield suitable specimens, and common mistakes to avoid during each part of the process. Finally, we include recommendations for obtaining the highest quality micrographs from prepared specimens with current microscope, detector, and support technology. PMID:27572723

  19. Cryo-EM Structure Determination Using Segmented Helical Image Reconstruction.

    PubMed

    Fromm, S A; Sachse, C

    2016-01-01

    Treating helices as single-particle-like segments followed by helical image reconstruction has become the method of choice for high-resolution structure determination of well-ordered helical viruses as well as flexible filaments. In this review, we will illustrate how the combination of latest hardware developments with optimized image processing routines have led to a series of near-atomic resolution structures of helical assemblies. Originally, the treatment of helices as a sequence of segments followed by Fourier-Bessel reconstruction revealed the potential to determine near-atomic resolution structures from helical specimens. In the meantime, real-space image processing of helices in a stack of single particles was developed and enabled the structure determination of specimens that resisted classical Fourier helical reconstruction and also facilitated high-resolution structure determination. Despite the progress in real-space analysis, the combination of Fourier and real-space processing is still commonly used to better estimate the symmetry parameters as the imposition of the correct helical symmetry is essential for high-resolution structure determination. Recent hardware advancement by the introduction of direct electron detectors has significantly enhanced the image quality and together with improved image processing procedures has made segmented helical reconstruction a very productive cryo-EM structure determination method. PMID:27572732

  20. Orthogonalizing EM: A design-based least squares algorithm

    PubMed Central

    Xiong, Shifeng; Dai, Bin; Huling, Jared; Qian, Peter Z. G.

    2016-01-01

    We introduce an efficient iterative algorithm, intended for various least squares problems, based on a design of experiments perspective. The algorithm, called orthogonalizing EM (OEM), works for ordinary least squares and can be easily extended to penalized least squares. The main idea of the procedure is to orthogonalize a design matrix by adding new rows and then solve the original problem by embedding the augmented design in a missing data framework. We establish several attractive theoretical properties concerning OEM. For the ordinary least squares with a singular regression matrix, an OEM sequence converges to the Moore-Penrose generalized inverse-based least squares estimator. For ordinary and penalized least squares with various penalties, it converges to a point having grouping coherence for fully aliased regression matrices. Convergence and the convergence rate of the algorithm are examined. Finally, we demonstrate that OEM is highly efficient for large-scale least squares and penalized least squares problems, and is considerably faster than competing methods when n is much larger than p. Supplementary materials for this article are available online. PMID:27499558

  1. Effects of flour conditioning on cannibalism of T. castaneum eggs and pupae

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Cannibalism is a very important factor regulating population dynamics of the red flour beetle. After several days of feeding, the flour becomes conditioned by the beetles, which can affect rates of cannibalism. Flour conditioning is caused by an accumulation of feces, pheromones, and ethylquinone, w...

  2. Coming apart at the seams: morphological evidence for pregnathal head capsule borders in adult Tribolium castaneum

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Cephalization and seamless fusion of the anterior body segments during development obscure the segmental origins of the insect head. Most of the visible seams are thought to reflect infolding for structural reinforcement rather than a merger of segmental or cuticular plate borders. Incomplete fusion...

  3. Signalling properties and pharmacological analysis of two sulfakinin receptors from the red flour beetle, Tribolium castaneum

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Sulfakinin is an insect neuropeptide that constitutes an important component of the complex network of hormonal and neural factors that regulate feeding and digestion. The key modulating functions of sulfakinin are mediated by binding and signaling via G-protein coupled receptors. Although a subst...

  4. Molecular and functional analyses of amino acid decarboxylases involved in cuticle tanning in Tribolium castaneum

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspartate 1-decarboxylase (ADC) and dopa decarboxylase (DDC) provide b–alanine and dopamine used in insect cuticle tanning. Beta-alanine is conjugated with dopamine to yield N-b-alanyldopamine (NBAD), a substrate for the phenoloxidase laccase that catalyzes the synthesis of cuticle protein cross-li...

  5. Expression of an endoglucanase from Tribolium castaneum (TcEG1) in Saccharomyces cerevisiae

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Insects are a largely unexploited resource in prospecting for novel cellulolytic enzymes to improve the current production of fuel ethanol from lignocellulosic biomass. Cost-effectiveness of lignocellulosic ethanol production is expected to increase by the combination of cellulose degradation (sacch...

  6. RNA interference and dietary inhibitors induce a similar compensation response in Tribolium castaneum larvae

    Technology Transfer Automated Retrieval System (TEKTRAN)

    RNAi uses the host’s cellular machinery to degrade and reduce gene expression and has become a way to functionally test a gene of interest. RNAi technology is being used to develop pest control strategies in the agricultural community because it can be applied to non-model systems. However, there ha...

  7. Genes related to mitochondrial functions are differentially expressed in phosphine-resistant and -susceptible Tribolium castaneum

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Phosphine is a valuable fumigant to control pest populations in stored grains and grain products. However, recent studies indicate a substantial increase in phosphine resistance in major stored-cereal pests worldwide. To understand the molecular bases of phosphine resistance in insects, we used RNA-...

  8. Residual efficacy of aerosols to control Tribolium castaneum and Tribolium confusum

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aerosol insecticides can be important components of insect management plans for mills, food warehouses, and processing plants. In the United States, synergized pyrethrin insecticide is used alone or combined with an insect growth regulator (IGR) insecticide, either methoprene or hydroprene. The pres...

  9. Monitoring Survivability and Infectivity of Porcine Epidemic Diarrhea Virus (PEDv) in the Infected On-Farm Earthen Manure Storages (EMS)

    PubMed Central

    Tun, Hein M.; Cai, Zhangbin; Khafipour, Ehsan

    2016-01-01

    In recent years, porcine epidemic diarrhea virus (PEDv) has caused major epidemics, which has been a burden to North America’s swine industry. Low infectious dose and high viability in the environment are major challenges in eradication of this virus. To further understand the viability of PEDv in the infected manure, we longitudinally monitored survivability and infectivity of PEDv in two open earthen manure storages (EMS; previously referred to as lagoon) from two different infected swine farms identified in the province of Manitoba, Canada. Our study revealed that PEDv could survive up to 9 months in the infected EMS after the initial outbreak in the farm. The viral load varied among different layers of the EMS with an average of 1.1 × 105 copies/ml of EMS, independent of EMS temperature and pH. In both studied EMS, the evidence of viral replication was observed through increased viral load in the later weeks of the samplings while there was no new influx of infected manure into the EMS, which was suggestive of presence of potential alternative hosts for PEDv within the EMS. Decreasing infectivity of virus over time irrespective of increased viral load suggested the possibility of PEDv evolution within the EMS and perhaps in the new host that negatively impacted virus infectivity. Viral load in the top layer of the EMS was low and mostly non-infective suggesting that environmental factors, such as UV and sunlight, could diminish the replicability and infectivity of the virus. Thus, frequent agitation of the EMS that could expose virus to UV and sunlight might be a potential strategy for reduction of PEDv load and infectivity in the infected EMS. PMID:27014197

  10. Overexpression of OsEm1 encoding a group I LEA protein confers enhanced drought tolerance in rice.

    PubMed

    Yu, Jing; Lai, Yongmin; Wu, Xi; Wu, Gang; Guo, Changkui

    2016-09-16

    Drought is the greatest threat for crops, including rice. In an effort to identify rice genes responsible for drought tolerance, a drought-responsive gene OsEm1 encoding a group I LEA protein, was chosen for this study. OsEm1 was shown at vegetative stages to be responsive to various abiotic stresses, including drought, salt, cold and the hormone ABA. In this study, we generated OsEm1-overexpressing rice plants to explore the function of OsEm1 under drought conditions. Overexpression of OsEm1 increases ABA sensitivity and enhances osmotic tolerance in rice. Compared with wild type, the OsEm1-overexpressing rice plants showed enhanced plant survival ratio at the vegetative stage; moreover, over expression of OsEm1 in rice increased the expression of other LEA genes, including RAB16A, RAB16C, RAB21, and LEA3, likely protecting organ integrity against harsh environments. Interestingly, the elevated level of OsEm1 had no different phenotype compared with wild type under normal condition. Our findings suggest that OsEm1 is a positive regulator of drought tolerance and is potentially promising for engineering drought tolerance in rice. PMID:27524243

  11. Prevention and Immunotherapy of Secondary Murine Alveolar Echinococcosis Employing Recombinant EmP29 Antigen

    PubMed Central

    Boubaker, Ghalia; Hemphill, Andrew; Huber, Cristina Olivia; Spiliotis, Markus; Babba, Hamouda; Gottstein, Bruno

    2015-01-01

    Alveolar echinococcosis (AE) is caused by infection with the larval stage of the tapeworm Echinococcus multilocularis. An increasing understanding of immunological events that account for the metacestode survival in human and murine AE infection prompted us to undertake explorative experiments tackling the potential of novel preventive and/or immunotherapeutic measures. In this study, the immunoprotective and immunotherapeutic ability of recombinant EmP29 antigen (rEmP29) was assessed in mice that were intraperitoneally infected with E. multilocularis metacestodes. For vaccination, three intraperitoneal injections with 20μg rEmP29 emulsified in saponin adjuvants were applied over 6 weeks. 2 weeks after the last boost, mice were infected, and at 90 days post-infection, rEmP29-vaccinated mice exhibited a median parasite weight that was reduced by 75% and 59% when compared to NaCl- or saponin–treated control mice, respectively. For immunotherapeutical application, the rEmP29 (20μg) vaccine was administered to experimentally infected mice, starting at 1 month post-infection, three times with 2 weeks intervals. Mice undergoing rEmP29 immunotherapy exhibited a median parasite load that was reduced by 53% and 49% when compared to NaCl- and saponin–treated control mice, respectively. Upon analysis of spleen cells, both, vaccination and treatment with rEmP29, resulted in low ratios of Th2/Th1 (IL-4/IFN-γ) cytokine mRNA and low levels of mRNA coding for IL-10 and IL-2. These results suggest that reduction of the immunosuppressive environment takes place in vaccinated as well as immunotreated mice, and a shift towards a Th1 type of immune response may be responsible for the observed increased restriction of parasite growth. The present study provides the first evidence that active immunotherapy may present a sustainable route for the control of AE. PMID:26053794

  12. EM-54 Technology Development In Situ Remediation Integrated Program. Annual report

    SciTech Connect

    Not Available

    1993-08-01

    The Department of Energy (DOE) established the Office of Technology Development (EM-50) as an element of Environmental Restoration and Waste Management (EM) in November 1989. EM manages remediation of all DOE sites as well as wastes from current operations. The goal of the EM program is to minimize risks to human health, safety and the environment, and to bring all DOE sites into compliance with Federal, state, and local regulations by 2019. EM-50 is charged with developing new technologies that are safer, more effective and less expensive than current methods. The In Situ Remediation Integrated Program (the subject of this report) is part of EM-541, the Environmental Restoration Research and Development Division of EM-54. The In Situ Remediation Integrated Program (ISR IP) was instituted out of recognition that in situ remediation could fulfill three important criteria: Significant cost reduction of cleanup by eliminating or minimizing excavation, transportation, and disposal of wastes; reduced health impacts on workers and the public by minimizing exposure to wastes during excavation and processing; and remediation of inaccessible sites, including: deep subsurfaces; in, under, and around buildings. Buried waste, contaminated soils and groundwater, and containerized wastes are all candidates for in situ remediation. Contaminants include radioactive wastes, volatile and non-volatile organics, heavy metals, nitrates, and explosive materials. The ISR IP tends to facilitate development of in situ remediation technologies for hazardous, radioactive, and mixed wastes in soils, groundwater, and storage tanks. Near-term focus is on containment of the wastes, with treatment receiving greater effort in future years.

  13. Assessing the environmental performance of construction materials testing using EMS: An Australian study.

    PubMed

    Dejkovski, Nick

    2016-10-01

    This paper reports the audit findings of the waste management practices at 30 construction materials testing (CMT) laboratories (constituting 4.6% of total accredited CMT laboratories at the time of the audit) that operate in four Australian jurisdictions and assesses the organisation's Environmental Management System (EMS) for indicators of progress towards sustainable development (SD). In Australia, waste indicators are 'priority indicators' of environmental performance yet the quality and availability of waste data is poor. National construction and demolition waste (CDW) data estimates are not fully disaggregated and the contribution of CMT waste (classified as CDW) to the national total CDW landfill burden is difficult to quantify. The environmental and human impacts of anthropogenic release of hazardous substances contained in CMT waste into the ecosphere can be measured by construing waste indicators from the EMS. An analytical framework for evaluating the EMS is developed to elucidate CMT waste indicators and assess these indicators against the principle of proportionality. Assessing against this principle allows for: objective evaluations of whether the environmental measures prescribed in the EMS are 'proportionate' to the 'desired' (subjective) level of protection chosen by decision-makers; and benchmarking CMT waste indicators against aspirational CDW targets set by each Australian jurisdiction included in the audit. Construed together, the EMS derived waste indicators and benchmark data provide a composite indicator of environmental performance and progress towards SD. The key audit findings indicate: CMT laboratories have a 'poor' environmental performance (and overall progress towards SD) when EMS waste data are converted into indicator scores and assessed against the principle of proportionality; CMT waste recycling targets are lower when benchmarked against jurisdictional CDW waste recovery targets; and no significant difference in the average

  14. OV-Wav: um novo pacote para análise multiescalar em astronomia

    NASA Astrophysics Data System (ADS)

    Pereira, D. N. E.; Rabaça, C. R.

    2003-08-01

    Wavelets e outras formas de análise multiescalar têm sido amplamente empregadas em diversas áreas do conhecimento, sendo reconhecidamente superiores a técnicas mais tradicionais, como as análises de Fourier e de Gabor, em certas aplicações. Embora a teoria dos wavelets tenha começado a ser elaborada há quase trinta anos, seu impacto no estudo de imagens astronômicas tem sido pequeno até bem recentemente. Apresentamos um conjunto de programas desenvolvidos ao longo dos últimos três anos no Observatório do Valongo/UFRJ que possibilitam aplicar essa poderosa ferramenta a problemas comuns em astronomia, como a remoção de ruído, a detecção hierárquica de fontes e a modelagem de objetos com perfis de brilho arbitrários em condições não ideais. Este pacote, desenvolvido para execução em plataforma IDL, teve sua primeira versão concluída recentemente e está sendo disponibilizado à comunidade científica de forma aberta. Mostramos também resultados de testes controlados ao quais submetemos os programas, com a sua aplicação a imagens artificiais, com resultados satisfatórios. Algumas aplicações astrofísicas foram estudadas com o uso do pacote, em caráter experimental, incluindo a análise da componente de luz difusa em grupos compactos de galáxias de Hickson e o estudo de subestruturas de nebulosas planetárias no espaço multiescalar.

  15. Designing ergonomic interventions for EMS workers: concept generation of patient-handling devices.

    PubMed

    Conrad, Karen M; Reichelt, Paul A; Lavender, Steven A; Gacki-Smith, Jessica; Hattle, Sally

    2008-11-01

    Fire service personnel and private ambulance paramedics suffer musculoskeletal injuries as they lift and carry patients while performing emergency medical services (EMS). Engineering changes, such as the design of new EMS patient-handling devices, offer a potential intervention opportunity for combating this problem. The purpose of this qualitative descriptive study was to generate beginning ideas for the design of new EMS patient-handling devices that were framed within the contextual reality of the end user firefighter/paramedics. Guided by an ecological model of musculoskeletal injuries in the fire service, focus groups were conducted with 25 firefighter/paramedics from 13 suburban fire departments. Based on their availability, participants were assigned to one of three groups with each group focusing on a different EMS patient-handling scenario. Each group participated in two focus group sessions: one session to brainstorm ideas for devices and a second session to validate sketches of their design ideas. The sketches were professionally drawn by an industrial designer who attended all focus group sessions. Sketches, photos, videotapes, and written transcripts were content analyzed to describe the phenomena of interest. The ideas centered on EMS devices for lateral transfers, bed-to-stairchair transfers, and stair descent transport, and served as the starting point for the development of EMS devices in subsequent phases of a mixed method research study. The outcomes of this study were an improved understanding of the contextual issues that need to be considered in designing EMS patient handling devices and a set of industrial design sketches that served as a starting point for subsequent development of the devices. End user acceptance criteria for the devices included: affordability, portability/compactness, durability, operability including being quickly ready for use, and cleanability.

  16. Assessing the environmental performance of construction materials testing using EMS: An Australian study.

    PubMed

    Dejkovski, Nick

    2016-10-01

    This paper reports the audit findings of the waste management practices at 30 construction materials testing (CMT) laboratories (constituting 4.6% of total accredited CMT laboratories at the time of the audit) that operate in four Australian jurisdictions and assesses the organisation's Environmental Management System (EMS) for indicators of progress towards sustainable development (SD). In Australia, waste indicators are 'priority indicators' of environmental performance yet the quality and availability of waste data is poor. National construction and demolition waste (CDW) data estimates are not fully disaggregated and the contribution of CMT waste (classified as CDW) to the national total CDW landfill burden is difficult to quantify. The environmental and human impacts of anthropogenic release of hazardous substances contained in CMT waste into the ecosphere can be measured by construing waste indicators from the EMS. An analytical framework for evaluating the EMS is developed to elucidate CMT waste indicators and assess these indicators against the principle of proportionality. Assessing against this principle allows for: objective evaluations of whether the environmental measures prescribed in the EMS are 'proportionate' to the 'desired' (subjective) level of protection chosen by decision-makers; and benchmarking CMT waste indicators against aspirational CDW targets set by each Australian jurisdiction included in the audit. Construed together, the EMS derived waste indicators and benchmark data provide a composite indicator of environmental performance and progress towards SD. The key audit findings indicate: CMT laboratories have a 'poor' environmental performance (and overall progress towards SD) when EMS waste data are converted into indicator scores and assessed against the principle of proportionality; CMT waste recycling targets are lower when benchmarked against jurisdictional CDW waste recovery targets; and no significant difference in the average

  17. Rock 'Em, Sock 'Em!

    ERIC Educational Resources Information Center

    Waters, John K.

    2011-01-01

    K-12-level competitive robotics is growing in popularity around the country and worldwide. According to one of the leading organizers of these events, FIRST--For Inspiration and Recognition of Science and Technology--250,000 students from 56 countries take part in its competitions. FIRST Tech Challenge (FTC) is a yearly event the organization puts…

  18. Pre-flight risk assessment in Emergency Medical Service (EMS) helicopters

    NASA Technical Reports Server (NTRS)

    Shively, R. J.

    1992-01-01

    The Emergency Medical Service (EMS) industry has been the subject of several television and newspaper articles (Harvey and Jensen, 1987) which emphasized the negative aspects, (e.g., fatalities and high accident rates), rather than the life saving services performed. Until recently, the accident rate of the EMS industry has been five times as high as that of other civil helicopters. This high accident rate has been coupled with the dramatic rise in the number of programs. The industry has built from a single service at its inception in 1972, to over 180 in 1987 (Spray, 1987), to the point that 93 percent of the contiguous U.S. is now covered by some type of EMS service. These factors prompted the National Transportation Safety Board (NTSB) to study the accidents that occurred between May 11, 1978 and December 3, 1986 (NTSB, 1988). The NTSB report concluded that 'Sound pilot judgment is central to safe flight operations.' They further stated that '... factors unique to EMS helicopter operations--such as the influence of the mission itself, program competition, and EMS program management perspectives--can drastically influence pilot judgment during the EMS mission.' One of the most difficult decisions that a pilot must make is whether to accept or decline a mission. A pre-flight risk assessment system (SAFE) was developed at NASA-Ames Research Center for civil EMS operations to aid pilots in making this decision objectively. The ability of the SAFE system to predict mission risk profiles was tested at an EMS facility. The results of this field study demonstrated that the usefulness of SAFE was highly dependent on the type of mission flown. SAFE is now being modified so that it can 'learn' with each mission flown. For example, after flying a mission to a particular site, an EMS pilot would input information about this mission into the system, such as new buildings, wires, or approach procedures. Then, the next time a pilot flew a similar mission or one to the same

  19. Does contamination buildup limit throughput for automated cryoEM?

    PubMed

    Cheng, Anchi; Fellmann, Denis; Pulokas, James; Potter, Clinton S; Carragher, Bridget

    2006-06-01

    The development of automated systems for data acquisition in cryo electron microscopy has enabled the possibility of acquiring very large number of images from a single specimen grid. We have demonstrated that over images of 250,000 single particles can be acquired in a 24 h period. This has raised questions as to whether contamination buildup on the specimen limits the quality of the data that can be acquired during these long duration experiments and also whether the data acquisition session could be extended to allow acquisition of more than 1,000,000 particles. We report here a systematic characterization of contamination of specimens maintained for long periods of time at liquid nitrogen temperatures using standard side entry cryo stages. As part of this characterization we developed a more reliable method for accurately estimating specimen ice thickness. Using the method, we were able to calibrate image contrast against ice thickness under a variety of magnifications, objective aperture positions, and defoci, and demonstrated the strong dependence of the calibration curve on these parameters. The results show the anti-contamination aperture is, as expected, critical to the prevention of contamination and that loading film into the microscope dramatically increases the contamination rate, particularly in the first 3 h after the insertion of the film box. In the absence of film, we were able to reproducibly demonstrate that the contamination rate can be limited to a rate of approximately 1 angstrom/h providing reassurance that contamination will not be a major limiting factor for long term cryoEM experiments if a CCD camera is used for the imaging.

  20. Elimination of error factors, affecting EM and seismic inversions

    NASA Astrophysics Data System (ADS)

    Magomedov, M.; Zuev, M. A.; Korneev, V. A.; Goloshubin, G.; Zuev, J.; Brovman, Y.

    2013-12-01

    EM or seismic data inversions are affected by many factors, which may conceal the responses from target objects. We address here the contributions from the following effects: 1) Pre-survey spectral sensitivity factor. Preliminary information about a target layer can be used for a pre-survey estimation of the required frequency domain and signal level. A universal approach allows making such estimations in real time, helping the survey crew to optimize an acquisition process. 2) Preliminary velocities' identification and their dispersions for all the seismic waves, arising in a stratified media became a fast working tool, based on the exact analytical solution. 3) Vertical gradients effect. For most layers the log data scatter, requiring an averaging pattern. A linear gradient within each representative layer is a reasonable compromise between required inversion accuracy and forward modeling complexity. 4) An effect from the seismic source's radial component becomes comparable with vertical part for explosive sources. If this effect is not taken into account, a serious modeling error takes place. This problem has an algorithmic solution. 5) Seismic modeling is often based on different representations for a source formulated either for a force or to a potential. The wave amplitudes depend on the formulation, making an inversion result sensitive to it. 6) Asymmetrical seismic waves (modified Rayleigh) in symmetrical geometry around liquid fracture come from S-wave and merge with the modified Krauklis wave at high frequencies. A detail analysis of this feature allows a spectral range optimization for the proper wave's extraction. 7) An ultrasonic experiment was conducted to show different waves appearance for a super-thin water-saturated fracture between two Plexiglas plates, being confirmed by comparison with theoretical computations. 8) A 'sandwich effect' was detected by comparison with averaged layer's effect. This opens an opportunity of the shale gas direct

  1. Incubator embedded cell culture imaging system (EmSight) based on Fourier ptychographic microscopy.

    PubMed

    Kim, Jinho; Henley, Beverley M; Kim, Charlene H; Lester, Henry A; Yang, Changhuei

    2016-08-01

    Multi-day tracking of cells in culture systems can provide valuable information in bioscience experiments. We report the development of a cell culture imaging system, named EmSight, which incorporates multiple compact Fourier ptychographic microscopes with a standard multiwell imaging plate. The system is housed in an incubator and presently incorporates six microscopes. By using the same low magnification objective lenses as the objective and the tube lens, the EmSight is configured as a 1:1 imaging system that, providing large field-of-view (FOV) imaging onto a low-cost CMOS imaging sensor. The EmSight improves the image resolution by capturing a series of images of the sample at varying illumination angles; the instrument reconstructs a higher-resolution image by using the iterative Fourier ptychographic algorithm. In addition to providing high-resolution brightfield and phase imaging, the EmSight is also capable of fluorescence imaging at the native resolution of the objectives. We characterized the system using a phase Siemens star target, and show four-fold improved coherent resolution (synthetic NA of 0.42) and a depth of field of 0.2 mm. To conduct live, long-term dopaminergic neuron imaging, we cultured ventral midbrain from mice driving eGFP from the tyrosine hydroxylase promoter. The EmSight system tracks movements of dopaminergic neurons over a 21 day period. PMID:27570701

  2. New Advances for a joint 3D inversion of multiple EM methods

    NASA Astrophysics Data System (ADS)

    Meqbel, N. M.; Ritter, O.

    2013-12-01

    Electromagnetic (EM) methods are routinely applied to image the subsurface from shallow to regional structures. Individual EM methods differ in their sensitivities towards resistive and conductive structures as well as in their exploration depths. Joint 3D inversion of multiple EM data sets can result in significantly better resolution of subsurface structures than the individual inversions. Proper weighting between different EM data is essential, however. We present a recently developed weighting algorithm to combine magnetotelluric (MT), controlled source EM (CSEM) and DC-geoelectric (DC) data. It is well known that MT data are mostly sensible to regional conductive structures, whereas, CSEM and DC data are more suitable to recover more shallow and resistive structures. Our new scheme is based on weighting individual components of the total data gradient after each model update. Norms of each data residual are used to assess how much weight individual components of the total data gradient must have to achieve an equal contribution of all data sets in the inverse model. A numerically efficient way to search for appropriate weighting factors could be established by applying a bi-diagonalization procedure to the sensitivity matrix. Thereby, the original inverse problem can be projected onto a smaller dimension in which the search of weighting factors is numerically cheap. We demonstrate the efficiency of the proposed weighting schemes and explore the model domain with synthetic data sets.

  3. dbEM: A database of epigenetic modifiers curated from cancerous and normal genomes

    PubMed Central

    Singh Nanda, Jagpreet; Kumar, Rahul; Raghava, Gajendra P. S.

    2016-01-01

    We have developed a database called dbEM (database of Epigenetic Modifiers) to maintain the genomic information of about 167 epigenetic modifiers/proteins, which are considered as potential cancer targets. In dbEM, modifiers are classified on functional basis and comprise of 48 histone methyl transferases, 33 chromatin remodelers and 31 histone demethylases. dbEM maintains the genomic information like mutations, copy number variation and gene expression in thousands of tumor samples, cancer cell lines and healthy samples. This information is obtained from public resources viz. COSMIC, CCLE and 1000-genome project. Gene essentiality data retrieved from COLT database further highlights the importance of various epigenetic proteins for cancer survival. We have also reported the sequence profiles, tertiary structures and post-translational modifications of these epigenetic proteins in cancer. It also contains information of 54 drug molecules against different epigenetic proteins. A wide range of tools have been integrated in dbEM e.g. Search, BLAST, Alignment and Profile based prediction. In our analysis, we found that epigenetic proteins DNMT3A, HDAC2, KDM6A, and TET2 are highly mutated in variety of cancers. We are confident that dbEM will be very useful in cancer research particularly in the field of epigenetic proteins based cancer therapeutics. This database is available for public at URL: http://crdd.osdd.net/raghava/dbem. PMID:26777304

  4. Single-Particle Cryo-EM of the Ryanodine Receptor Channel in an Aqueous Environment.

    PubMed

    Baker, Mariah R; Fan, Guizhen; Serysheva, Irina I

    2015-01-01

    Ryanodine receptors (RyRs) are tetrameric ligand-gated Ca(2+) release channels that are responsible for the increase of cytosolic Ca(2+) concentration leading to muscle contraction. Our current understanding of RyR channel gating and regulation is greatly limited due to the lack of a high-resolution structure of the channel protein. The enormous size and unwieldy shape of Ca(2+) release channels make X-ray or NMR methods difficult to apply for high-resolution structural analysis of the full-length functional channel. Single-particle electron cryo-microscopy (cryo-EM) is one of the only effective techniques for the study of such a large integral membrane protein and its molecular interactions. Despite recent developments in cryo-EM technologies and break-through single-particle cryo-EM studies of ion channels, cryospecimen preparation, particularly the presence of detergent in the buffer, remains the main impediment to obtaining atomic-resolution structures of ion channels and a multitude of other integral membrane protein complexes. In this review we will discuss properties of several detergents that have been successfully utilized in cryo-EM studies of ion channels and the emergence of the detergent alternative amphipol to stabilize ion channels for structure-function characterization. Future structural studies of challenging specimen like ion channels are likely to be facilitated by cryo-EM amenable detergents or alternative surfactants.

  5. Description and comparison of algorithms for correcting anisotropic magnification in cryo-EM images.

    PubMed

    Zhao, Jianhua; Brubaker, Marcus A; Benlekbir, Samir; Rubinstein, John L

    2015-11-01

    Single particle electron cryomicroscopy (cryo-EM) allows for structures of proteins and protein complexes to be determined from images of non-crystalline specimens. Cryo-EM data analysis requires electron microscope images of randomly oriented ice-embedded protein particles to be rotated and translated to allow for coherent averaging when calculating three-dimensional (3D) structures. Rotation of 2D images is usually done with the assumption that the magnification of the electron microscope is the same in all directions. However, due to electron optical aberrations, this condition is not met with some electron microscopes when used with the settings necessary for cryo-EM with a direct detector device (DDD) camera. Correction of images by linear interpolation in real space has allowed high-resolution structures to be calculated from cryo-EM images for symmetric particles. Here we describe and compare a simple real space method, a simple Fourier space method, and a somewhat more sophisticated Fourier space method to correct images for a measured anisotropy in magnification. Further, anisotropic magnification causes contrast transfer function (CTF) parameters estimated from image power spectra to have an apparent systematic astigmatism. To address this problem we develop an approach to adjust CTF parameters measured from distorted images so that they can be used with corrected images. The effect of anisotropic magnification on CTF parameters provides a simple way of detecting magnification anisotropy in cryo-EM datasets.

  6. Cryo-EM studies of the structure and dynamics of vacuolar-type ATPases.

    PubMed

    Mazhab-Jafari, Mohammad T; Rubinstein, John L

    2016-07-01

    Electron cryomicroscopy (cryo-EM) has significantly advanced our understanding of molecular structure in biology. Recent innovations in both hardware and software have made cryo-EM a viable alternative for targets that are not amenable to x-ray crystallography or nuclear magnetic resonance (NMR) spectroscopy. Cryo-EM has even become the method of choice in some situations where x-ray crystallography and NMR spectroscopy are possible but where cryo-EM can determine structures at higher resolution or with less time or effort. Rotary adenosine triphosphatases (ATPases) are crucial to the maintenance of cellular homeostasis. These enzymes couple the synthesis or hydrolysis of adenosine triphosphate to the use or production of a transmembrane electrochemical ion gradient, respectively. However, the membrane-embedded nature and conformational heterogeneity of intact rotary ATPases have prevented their high-resolution structural analysis to date. Recent application of cryo-EM methods to the different types of rotary ATPase has led to sudden advances in understanding the structure and function of these enzymes, revealing significant conformational heterogeneity and characteristic transmembrane α helices that are highly tilted with respect to the membrane. In this Review, we will discuss what has been learned recently about rotary ATPase structure and function, with a particular focus on the vacuolar-type ATPases.

  7. Description and comparison of algorithms for correcting anisotropic magnification in cryo-EM images.

    PubMed

    Zhao, Jianhua; Brubaker, Marcus A; Benlekbir, Samir; Rubinstein, John L

    2015-11-01

    Single particle electron cryomicroscopy (cryo-EM) allows for structures of proteins and protein complexes to be determined from images of non-crystalline specimens. Cryo-EM data analysis requires electron microscope images of randomly oriented ice-embedded protein particles to be rotated and translated to allow for coherent averaging when calculating three-dimensional (3D) structures. Rotation of 2D images is usually done with the assumption that the magnification of the electron microscope is the same in all directions. However, due to electron optical aberrations, this condition is not met with some electron microscopes when used with the settings necessary for cryo-EM with a direct detector device (DDD) camera. Correction of images by linear interpolation in real space has allowed high-resolution structures to be calculated from cryo-EM images for symmetric particles. Here we describe and compare a simple real space method, a simple Fourier space method, and a somewhat more sophisticated Fourier space method to correct images for a measured anisotropy in magnification. Further, anisotropic magnification causes contrast transfer function (CTF) parameters estimated from image power spectra to have an apparent systematic astigmatism. To address this problem we develop an approach to adjust CTF parameters measured from distorted images so that they can be used with corrected images. The effect of anisotropic magnification on CTF parameters provides a simple way of detecting magnification anisotropy in cryo-EM datasets. PMID:26087140

  8. Incubator embedded cell culture imaging system (EmSight) based on Fourier ptychographic microscopy

    PubMed Central

    Kim, Jinho; Henley, Beverley M.; Kim, Charlene H.; Lester, Henry A.; Yang, Changhuei

    2016-01-01

    Multi-day tracking of cells in culture systems can provide valuable information in bioscience experiments. We report the development of a cell culture imaging system, named EmSight, which incorporates multiple compact Fourier ptychographic microscopes with a standard multiwell imaging plate. The system is housed in an incubator and presently incorporates six microscopes. By using the same low magnification objective lenses as the objective and the tube lens, the EmSight is configured as a 1:1 imaging system that, providing large field-of-view (FOV) imaging onto a low-cost CMOS imaging sensor. The EmSight improves the image resolution by capturing a series of images of the sample at varying illumination angles; the instrument reconstructs a higher-resolution image by using the iterative Fourier ptychographic algorithm. In addition to providing high-resolution brightfield and phase imaging, the EmSight is also capable of fluorescence imaging at the native resolution of the objectives. We characterized the system using a phase Siemens star target, and show four-fold improved coherent resolution (synthetic NA of 0.42) and a depth of field of 0.2 mm. To conduct live, long-term dopaminergic neuron imaging, we cultured ventral midbrain from mice driving eGFP from the tyrosine hydroxylase promoter. The EmSight system tracks movements of dopaminergic neurons over a 21 day period. PMID:27570701

  9. Cryo-EM studies of the structure and dynamics of vacuolar-type ATPases.

    PubMed

    Mazhab-Jafari, Mohammad T; Rubinstein, John L

    2016-07-01

    Electron cryomicroscopy (cryo-EM) has significantly advanced our understanding of molecular structure in biology. Recent innovations in both hardware and software have made cryo-EM a viable alternative for targets that are not amenable to x-ray crystallography or nuclear magnetic resonance (NMR) spectroscopy. Cryo-EM has even become the method of choice in some situations where x-ray crystallography and NMR spectroscopy are possible but where cryo-EM can determine structures at higher resolution or with less time or effort. Rotary adenosine triphosphatases (ATPases) are crucial to the maintenance of cellular homeostasis. These enzymes couple the synthesis or hydrolysis of adenosine triphosphate to the use or production of a transmembrane electrochemical ion gradient, respectively. However, the membrane-embedded nature and conformational heterogeneity of intact rotary ATPases have prevented their high-resolution structural analysis to date. Recent application of cryo-EM methods to the different types of rotary ATPase has led to sudden advances in understanding the structure and function of these enzymes, revealing significant conformational heterogeneity and characteristic transmembrane α helices that are highly tilted with respect to the membrane. In this Review, we will discuss what has been learned recently about rotary ATPase structure and function, with a particular focus on the vacuolar-type ATPases. PMID:27532044

  10. Cryo-EM studies of the structure and dynamics of vacuolar-type ATPases

    PubMed Central

    Mazhab-Jafari, Mohammad T.; Rubinstein, John L.

    2016-01-01

    Electron cryomicroscopy (cryo-EM) has significantly advanced our understanding of molecular structure in biology. Recent innovations in both hardware and software have made cryo-EM a viable alternative for targets that are not amenable to x-ray crystallography or nuclear magnetic resonance (NMR) spectroscopy. Cryo-EM has even become the method of choice in some situations where x-ray crystallography and NMR spectroscopy are possible but where cryo-EM can determine structures at higher resolution or with less time or effort. Rotary adenosine triphosphatases (ATPases) are crucial to the maintenance of cellular homeostasis. These enzymes couple the synthesis or hydrolysis of adenosine triphosphate to the use or production of a transmembrane electrochemical ion gradient, respectively. However, the membrane-embedded nature and conformational heterogeneity of intact rotary ATPases have prevented their high-resolution structural analysis to date. Recent application of cryo-EM methods to the different types of rotary ATPase has led to sudden advances in understanding the structure and function of these enzymes, revealing significant conformational heterogeneity and characteristic transmembrane α helices that are highly tilted with respect to the membrane. In this Review, we will discuss what has been learned recently about rotary ATPase structure and function, with a particular focus on the vacuolar-type ATPases. PMID:27532044

  11. EM algorithm applied for estimating non-stationary region boundaries using electrical impedance tomography

    NASA Astrophysics Data System (ADS)

    Khambampati, A. K.; Rashid, A.; Kim, B. S.; Liu, Dong; Kim, S.; Kim, K. Y.

    2010-04-01

    EIT has been used for the dynamic estimation of organ boundaries. One specific application in this context is the estimation of lung boundaries during pulmonary circulation. This would help track the size and shape of lungs of the patients suffering from diseases like pulmonary edema and acute respiratory failure (ARF). The dynamic boundary estimation of the lungs can also be utilized to set and control the air volume and pressure delivered to the patients during artificial ventilation. In this paper, the expectation-maximization (EM) algorithm is used as an inverse algorithm to estimate the non-stationary lung boundary. The uncertainties caused in Kalman-type filters due to inaccurate selection of model parameters are overcome using EM algorithm. Numerical experiments using chest shaped geometry are carried out with proposed method and the performance is compared with extended Kalman filter (EKF). Results show superior performance of EM in estimation of the lung boundary.

  12. Direct and indirect measures of speech articulator motions using low power EM sensors

    SciTech Connect

    Barnes, T; Burnett, G; Gable, T; Holzrichter, J F; Ng, L

    1999-05-12

    Low power Electromagnetic (EM) Wave sensors can measure general properties of human speech articulator motions, as speech is produced. See Holzrichter, Burnett, Ng, and Lea, J.Acoust.Soc.Am. 103 (1) 622 (1998). Experiments have demonstrated extremely accurate pitch measurements (< 1 Hz per pitch cycle) and accurate onset of voiced speech. Recent measurements of pressure-induced tracheal motions enable very good spectra and amplitude estimates of a voiced excitation function. The use of the measured excitation functions and pitch synchronous processing enable the determination of each pitch cycle of an accurate transfer function and, indirectly, of the corresponding articulator motions. In addition, direct measurements have been made of EM wave reflections from articulator interfaces, including jaw, tongue, and palate, simultaneously with acoustic and glottal open/close signals. While several types of EM sensors are suitable for speech articulator measurements, the homodyne sensor has been found to provide good spatial and temporal resolution for several applications.

  13. Evaluation model calculations with the water reactor analysis package (WRAP-EM)

    SciTech Connect

    Gregory, M.V.; Beranek, F.

    1982-01-01

    The Water Reactor Analysis Package-Evaluation Model (WRAP-EM) is a modular system of computer codes designed to provide the safety analyst with the capability of performing complete loss-of-coolant calculations for both pressurized- and boiling-water reactor systems. The system provides a licensing-type calculation capability and thus contains most of the Nuclear Regulatory Commission-Approved EM options, as described in the Code of Federal Regulations, Title 10, Part 50, Appendix K. All phases of an accident (blowdown, refill, and reflood) are modeled. The WRAP consists of modified versions of five preexisting codes (RELAP4/MOD5, GAPCON, FRAP, MOXY, and NORCOOL), the necessary interfaces to permit automatic transition from one code to the next during the transient calculations, plus a host of user-convenience features to aid the analyst faced with a multitude of EM calculations. The WRAP has been verified against both calculated and experimental results.

  14. Physics-based simulation of EM and SM in TSV-based 3D IC structures

    SciTech Connect

    Kteyan, Armen; Sukharev, Valeriy; Zschech, Ehrenfried

    2014-06-19

    Evolution of stresses in through-silicon-vias (TSVs) and in the TSV landing pad due to the stress migration (SM) and electromigration (EM) phenomena are considered. It is shown that an initial stress distribution existing in a TSV depends on its architecture and copper fill technology. We demonstrate that in the case of proper copper annealing the SM-induced redistribution of atoms results in uniform distributions of the hydrostatic stress and concentration of vacancies along each segment. In this case, applied EM stressing generates atom migration that is characterized by kinetics depending on the preexisting equilibrium concentration of vacancies. Stress-induced voiding in TSV is considered. EM induced voiding in TSV landing pad is analyzed in details.

  15. An Introduction to Emergency Medical Services (EMS). Pre-Hospital Phase. Emergency Medical Services Orientation, Lesson Plan No. 9.

    ERIC Educational Resources Information Center

    Young, Derrick P.

    Designed for use with interested students at high schools, community colleges, and four-year colleges, this lesson plan was developed to provide an introduction to the pre-hospital phase of Emergency Medical Services (EMS) and to serve as a recruitment tool for the EMS Program at Kapiolani Community College (KCC) in Hawaii. The objectives of the…

  16. Factor Analysis with EM Algorithm Never Gives Improper Solutions when Sample Covariance and Initial Parameter Matrices Are Proper

    ERIC Educational Resources Information Center

    Adachi, Kohei

    2013-01-01

    Rubin and Thayer ("Psychometrika," 47:69-76, 1982) proposed the EM algorithm for exploratory and confirmatory maximum likelihood factor analysis. In this paper, we prove the following fact: the EM algorithm always gives a proper solution with positive unique variances and factor correlations with absolute values that do not exceed one, when the…

  17. Mass Gathering Medical Care: Resource Document for the National Association of EMS Physicians Position Statement.

    PubMed

    Schwartz, Brian; Nafziger, Sarah; Milsten, Andrew; Luk, Jeffrey; Yancey, Arthur

    2015-01-01

    Mass gatherings are heterogeneous in terms of size, duration, type of event, crowd behavior, demographics of the participants and spectators, use of recreational substances, weather, and environment. The goals of health and medical services should be the provision of care for participants and spectators consistent with local standards of care, protection of continuing medical service to the populations surrounding the event venue, and preparation for surge to respond to extraordinary events. Pre-event planning among jurisdictional public health and EMS, acute care hospitals, and event EMS is essential, but should also include, at a minimum, event security services, public relations, facility maintenance, communications technicians, and the event planners and organizers. Previous documented experience with similar events has been shown to most accurately predict future needs. Future work in and guidance for mass gathering medical care should include the consistent use and further development of universally accepted consistent metrics, such as Patient Presentation Rate and Transfer to Hospital Rate. Only by standardizing data collection can evaluations be performed that link interventions with outcomes to enhance evidence-based EMS services at mass gatherings. Research is needed to evaluate the skills and interventions required by EMS providers to achieve desired outcomes. The event-dedicated EMS Medical Director is integral to acceptable quality medical care provided at mass gatherings; hence, he/she must be included in all aspects of mass gathering medical care planning, preparations, response, and recovery. Incorporation of jurisdictional EMS and community hospital medical leadership, and emergency practitioners into these processes will ensure that on-site care, transport, and transition to acute care at appropriate receiving facilities is consistent with, and fully integrated into the community's medical care system, while fulfilling the needs of event

  18. Análise da medição do raio solar em ultravioleta

    NASA Astrophysics Data System (ADS)

    Saraiva, A. C. V.; Giménez de Castro, C. G.; Costa, J. E. R.; Selhorst, C. L.; Simões, P. J. A.

    2003-08-01

    A medição acurada do raio solar em qualquer banda do espectro eletromagnético é de relevância na formulação e calibração de modelos da estrutura e atmosfera solar. Esses modelos atribuem emissão do contínuo do Sol calmo em microondas à mesma região da linha Ha do Hell. Apresentamos a medição do raio solar em UV com imagens do EIT (Extreme Ultraviolet Image Telescope) entre 1996 e 2002, no comprimento de onda 30,9 nm (Ha do Hell), que se forma na região de transição/cromosfera solar. A técnica utilizada para o cálculo do raio UV foi baseada na transformada Wavelet B3spline. Fizemos um banco de dados com 1 imagem por dia durante o período citado. Obtivemos como resultado o raio médio da ordem de 975.61" e uma diminuição do mesmo para o período citado variando em média -0,45" /ano. Comparamos estes dados com os valores obtidos pelo ROI (Radio Observatório de Itapetinga) em 22/48 GHz e Nobeyama Radio Heliograph em 17 GHz mostrando que os raios médios são muito próximos o que indica que a região de formação nessas freqüências é a mesma conforme os modelos. Comparamos os resultados também com outros índices de atividade solar.

  19. Recidivism Among Licensed-Released Prisoners Who Participated in the EM Program in Israel.

    PubMed

    Shoham, Efrat; Yehosha-Stern, Shirley; Efodi, Rotem

    2015-08-01

    Toward the end of 2006, a pilot program was launched in Israel wherein licensed-released prisoners were put under electronic monitoring (EM). In addition to EM, the pilot program, operated by the Prisoners' Rehabilitation Authority, provides programs of occupational supervision and personal therapy and is designed to allow for early release of those prisoners who, without increased supervision, would have been found unsuitable for early release. The aim of this study was to ascertain whether participation in the EM program among licensed-released prisoners in Israel might bring about lessened recidivism. For that matter, rates of arrests and incarceration were examined during a follow-up period of up to 4 years, among the entirety of licensed-released prisoners participating in the EM program between the years 2007 and 2009 (n = 155). To compare recidivism rates, a control group was assembled from among the entirety of released prisoners who were found unsuitable for early release in judicial conditions, and had therefore served the full term of their incarceration, to be released between the years 2005 and 2006 (a period of time during which an EM program was not yet operated among licensed-released prisoners in Israel). Study findings clearly show that while among the control group, 42% of released prisoners were re-incarcerated, at the end of a 4-year follow-up period, only 15% among the study group had returned to prison. These findings can be explained by combining the Social Control theory and the Self-Control theory which consider the period of time under EM program and the occupational and familial integration tools for reducing criminal connections and enhancing pro-social behavior. PMID:24510371

  20. Influence of motivations for seeking ISO 14001 certification on perceptions of EMS effectiveness in China.

    PubMed

    Fryxell, Gerald E; Lo, Carlos Wing-Hung; Chung, Shan Shan

    2004-02-01

    This study examines the motivations of mainland Chinese facilities in seeking ISO 14001 certification and examines the linkages between these motivations and self-reports of the effectiveness of major environmental management system (EMS) components. In a sample of 128 facilities in Beijing, Shanghai, and Guangzhou, the main drivers for certification were reported to be to ensure regulatory compliance, to enhance the firm's reputation, and to improve environmental performance, in that order. Although motivation to achieve cost reductions were least emphasized, a broad range of motivations appears to be considered in the decision to seek certification to ISO 14001. Regression models linking these motivations to the EMS components suggests that internal motivations have an influence on most EMS components. One interesting exception to this, however, is that no significant relationship was observed between internal motivations and the promulgation of environmental objectives and targets. The relationships associated with external motivations for certification (i.e., those in response to customer and other stakeholder pressures) and EMS components, on the other hand, are weaker and tend to occur earlier in the process cycle. No significant relationships were found between motivations to reduce costs and perceptions of the effectiveness of EMS components. Overall, these findings suggest that ISO 14001, as currently being implemented in mainland China, may have a modestly useful role when used in combination with other policy mechanisms to move the Chinese economy toward more sustainable practices. It is asserted that the ISO standard could provide even greater benefits if Chinese registrars were more proactive in developing EMS in conjunction with even more rigorous third-party audits.

  1. Mass Gathering Medical Care: Resource Document for the National Association of EMS Physicians Position Statement.

    PubMed

    Schwartz, Brian; Nafziger, Sarah; Milsten, Andrew; Luk, Jeffrey; Yancey, Arthur

    2015-01-01

    Mass gatherings are heterogeneous in terms of size, duration, type of event, crowd behavior, demographics of the participants and spectators, use of recreational substances, weather, and environment. The goals of health and medical services should be the provision of care for participants and spectators consistent with local standards of care, protection of continuing medical service to the populations surrounding the event venue, and preparation for surge to respond to extraordinary events. Pre-event planning among jurisdictional public health and EMS, acute care hospitals, and event EMS is essential, but should also include, at a minimum, event security services, public relations, facility maintenance, communications technicians, and the event planners and organizers. Previous documented experience with similar events has been shown to most accurately predict future needs. Future work in and guidance for mass gathering medical care should include the consistent use and further development of universally accepted consistent metrics, such as Patient Presentation Rate and Transfer to Hospital Rate. Only by standardizing data collection can evaluations be performed that link interventions with outcomes to enhance evidence-based EMS services at mass gatherings. Research is needed to evaluate the skills and interventions required by EMS providers to achieve desired outcomes. The event-dedicated EMS Medical Director is integral to acceptable quality medical care provided at mass gatherings; hence, he/she must be included in all aspects of mass gathering medical care planning, preparations, response, and recovery. Incorporation of jurisdictional EMS and community hospital medical leadership, and emergency practitioners into these processes will ensure that on-site care, transport, and transition to acute care at appropriate receiving facilities is consistent with, and fully integrated into the community's medical care system, while fulfilling the needs of event

  2. EMS transports patients to clinics--seeks to relieve ED crowding.

    PubMed

    2010-07-01

    With most EDs overcrowded and with no relief in sight, ED managers will take all the help they can get. In some areas of the country, the local EMS providers are looking to offer some relief by transporting less urgent patients to alternative sites such as urgent care clinics. EDs assist in the development of program protocols and processes. ED nurses are considered for consultation on whether a patient can appropriately be seen in a less urgent setting. EMS providers must be trained on the requirements patients must fulfill to be transported to an ED PMID:20608479

  3. Denoising of human speech using combined acoustic and em sensor signal processing

    SciTech Connect

    Ng, L C; Burnett, G C; Holzrichter, J F; Gable, T J

    1999-11-29

    Low Power EM radar-like sensors have made it possible to measure properties of the human speech production system in real-time, without acoustic interference. This greatly enhances the quality and quantify of information for many speech related applications. See Holzrichter, Burnett, Ng, and Lea, J. Acoustic. Soc. Am. 103 (1) 622 (1998). By using combined Glottal-EM- Sensor- and Acoustic-signals, segments of voiced, unvoiced, and no-speech can be reliably defined. Real-time Denoising filters can be constructed to remove noise from the user's corresponding speech signal.

  4. Label-Free Visualization of Ultrastructural Features of Artificial Synapses via Cryo-EM

    PubMed Central

    2011-01-01

    The ultrastructural details of presynapses formed between artificial substrates of submicrometer silica beads and hippocampal neurons are visualized via cryo-electron microscopy (cryo-EM). The silica beads are derivatized by poly-d-lysine or lipid bilayers. Molecular features known to exist at presynapses are clearly present at these artificial synapses, as visualized by cryo-EM. Key synaptic features such as the membrane contact area at synaptic junctions, the presynaptic bouton containing presynaptic vesicles, as well as microtubular structures can be identified. This is the first report of the direct, label-free observation of ultrastructural details of artificial synapses. PMID:22860164

  5. Use of the AIC with the EM algorithm: A demonstration of a probability model selection technique

    SciTech Connect

    Glosup, J.G.; Axelrod M.C.

    1994-11-15

    The problem of discriminating between two potential probability models, a Gaussian distribution and a mixture of Gaussian distributions, is considered. The focus of our interest is a case where the models are potentially non-nested and the parameters of the mixture model are estimated through the EM algorithm. The AIC, which is frequently used as a criterion for discriminating between non-nested models, is modified to work with the EM algorithm and is shown to provide a model selection tool for this situation. A particular problem involving an infinite mixture distribution known as Middleton`s Class A model is used to demonstrate the effectiveness and limitations of this method.

  6. Rotating-bed reactor as a power source for EM gun applications

    SciTech Connect

    Powell, J.; Botts, T.; Stickley, C.M.; Meth, S.

    1980-01-01

    Electromagnetic gun applications of the Rotating Bed Reactor (RBR) are examined. The RBR is a compact (approx. 1 m/sup 3/), (up to several thousand MW(th)), high-power reactor concept, capable of producing a high-temperature (up to approx. 300/sup 0/K) gas stream with a MHD generator coupled to it, the RBR can generate electric power (up to approx. 1000 MW(e)) in the pulsed or cw modes. Three EM gun applications are investigated: a rail gun thruster for orbit transfer, a rapid-fire EM gun for point defense, and a direct ground-to-space launch. The RBR appears suitable for all applications.

  7. Study on rheo-diecasting process of 7075R alloys by SA-EMS melt homogenized treatment

    NASA Astrophysics Data System (ADS)

    Zhihua, G.; Jun, X.; Zhifeng, Z.; Guojun, L.; Mengou, T.

    2016-03-01

    An advanced melt processing technology, spiral annular electromagnetic stirring (SA-EMS) based on the annular electromagnetic stirring (A-EMS) process was developed for manufacturing Al-alloy components with high integrity. The SA-EMS process innovatively combines non-contact electromagnetic stirring and a spiral annular chamber with specially designed profiles to in situ make high quality melt slurry, and intensive forced shearing can be achieved under high shear rate and high intensity of turbulence inside the spiral annular chamber. In this paper, the solidification microstructure and hardness of 7075R alloy die-casting connecting rod conditioned by the SA-EMS melt processing technology were investigated. The results indicate that, the SA-EMS melt processing technology exhibited superior grain refinement and remarkable structure homogeneity. In addition, it can evidently enhance the mechanical performance and reduce the crack tendency.

  8. Detecção inesperada de efeitos de lentes fracas em grupos de galáxias pouco luminosos em raios-X

    NASA Astrophysics Data System (ADS)

    Carrasco, R.; Mendes de Oliveira, C.; Sodrã©, L., Jr.; Lima Neto, G. B.; Cypriano, E. S.; Lengruber, L. L.; Cuevas, H.; Ramirez, A.

    2003-08-01

    Obtivemos, como parte do programa de verificação científica do GMOS Sul, imagens profundas de três grupos de galáxias: G97 e G102 (z~0,4) e G124 (z = 0,17). Esses alvos foram selecionados a partir do catálogo de fontes extensas de Vikhlinin (1998), por terem luminosidades em raios X menores que 3´1043 ergs s-1, valor cerca de uma ou duas ordens de grandeza inferior ao de aglomerados de galáxias. O objetivo primário dessas observações é o estudo da evolução de galáxias em grupos. Grupos são ambientes menos densos que aglomerados, contêm a grande maioria das galáxias do Universo mas que, até o momento, foram estudados detalhadamente apenas no Universo local (z~0). Com esses dados efetuamos uma análise estatística da distorção na forma das galáxias de fundo (lentes gravitacionais fracas) como forma de inferir o conteúdo e a distribuição de massa nesses grupos apesar de que, em princípio, esse efeito não deveria ser detectado uma vez que os critérios de seleção adotados previlegiam sistemas de baixa massa. De fato, para G124 obtivemos apenas um limite superior para sua massa que é compatível com sua luminosidade em raios X. De modo contrário e surpreendente, os objetos G102 e G097, aparentam ter massas que resultariam em dispersões de velocidade maiores que 1000 km s-1, muito maiores do que se espera para grupos de galáxias. Com efeito, para G097 obtivemos, a partir de dados do satélite XMM, uma estimativa para a temperatura do gás intragrupo de kT = 2,6 keV, que é tipica de sistemas com dispersões de velocidade de ~ 600 km s-1, bem característica de grupos. Essas contradições aparentes entre lentes fracas e raios X podem ser explicadas de dois modos: i) a massa obtida por lentes estaria sobreestimada devido à superposição de estruturas massivas ao longo da linha de visada ou ii) a temperatura do gás do meio intra-grupo reflete o potencial gravitacional de estruturas menores que estariam se fundindo para formar uma

  9. 6. PHOTOCOPY, FOUNDATION AND FLOOR PLAN DRAWING OF E.M. BARRACKS, ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    6. PHOTOCOPY, FOUNDATION AND FLOOR PLAN DRAWING OF E.M. BARRACKS, N.C.O. AND OFFICERS QUARTERS. - NIKE Missile Base SL-40, Barracks No. 1, North end of base, southest of Basketball Court & northwest of Barracks No. 2, Hecker, Monroe County, IL

  10. 3D time-domain airborne EM modeling for an arbitrarily anisotropic earth

    NASA Astrophysics Data System (ADS)

    Yin, Changchun; Qi, Yanfu; Liu, Yunhe

    2016-08-01

    Time-domain airborne EM data is currently interpreted based on an isotropic model. Sometimes, it can be problematic when working in the region with distinct dipping stratifications. In this paper, we simulate the 3D time-domain airborne EM responses over an arbitrarily anisotropic earth with topography by edge-based finite-element method. Tetrahedral meshes are used to describe the abnormal bodies with complicated shapes. We further adopt the Backward Euler scheme to discretize the time-domain diffusion equation for electric field, obtaining an unconditionally stable linear equations system. We verify the accuracy of our 3D algorithm by comparing with 1D solutions for an anisotropic half-space. Then, we switch attentions to effects of anisotropic media on the strengths and the diffusion patterns of time-domain airborne EM responses. For numerical experiments, we adopt three typical anisotropic models: 1) an anisotropic anomalous body embedded in an isotropic half-space; 2) an isotropic anomalous body embedded in an anisotropic half-space; 3) an anisotropic half-space with topography. The modeling results show that the electric anisotropy of the subsurface media has big effects on both the strengths and the distribution patterns of time-domain airborne EM responses; this effect needs to be taken into account when interpreting ATEM data in areas with distinct anisotropy.

  11. Lessons in Generative Design, Publishing, and Circulation: What EM-Journal's First Year Has Taught Us

    ERIC Educational Resources Information Center

    Rosinski, Jana; Lonsdale, Chelsea; Morrison, Becky; Mueller, Derek; Nannini, Adam

    2013-01-01

    "EM-Journal" is a flexibly refereed online journal featuring writing produced by students of Eastern Michigan University. The journal showcases a variety of documents (articles, essays, reports, etc.) written and designed by students enrolled in EMU's First-Year Writing (FYW) program, in selected Writing Intensive (WI) courses affiliated with the…

  12. Are you under stress in EMS. Understanding the slippery slope of burnout and PTSD.

    PubMed

    Collopy, Kevin T; Kivlehan, Sean M; Snyder, Scott R

    2012-10-01

    Burnout and PTSD are closely linked and often underreported in EMS. EMS classrooms do little or nothing to prepare providers for the inherent emotional stresses of emergency response and the "thick skin" culture of EMS may make many providers apprehensive about sharing their true feelings. Burnout is triggered by many of the same stresses that lead to the symptoms of PTSD and providers experiencing burnout that doesn't resolve within a few weeks may actually be experiencing PTSD. Be mindful of yourself and your fellow coworkers, particularly after a very traumatic response. And remember traumatic responses don't need to be as dramatic as Sept. 11, New Orleans after Hurricane Katrina or the Aurora, CO shootings to bother an EMS worker. In contrast, these are the calls where providers often receive the most attention. Instead, watch for the new father who just performed CPR on an infant the same age as his own, or the provider who just watched his or her friend die following a motor vehicle collision. Pay attention to yourself and colleagues, and be responsible and honest with yourself and others about when coping strategies are enough, and when they aren't. Finally, don't ever be afraid to seek help.

  13. New Jersey's EMS response to Superstorm Sandy: a case study of the emergency management assistance compact.

    PubMed

    Clancy, Terry; Christensen, Kenneth; Cortacans, Henry P

    2014-06-01

    In the United States, understanding the Emergency Management Assistance Compact (EMAC) is critical to responding to a natural disaster or manmade event. Recently, the State of New Jersey responded to Superstorm Sandy and implemented the EMAC system by requesting ambulances to aid in the Emergency Medical Services response. New Jersey's response to Superstorm Sandy was unprecedented in that this storm affected the entire state and EMS community. New Jersey's EMS community and infrastructure were impacted greatly, despite years of planning and preparation for such an event. Once received, out-of-state EMS resources were integrated into New Jersey's emergency management and EMS systems. In this report, each phase of the EMAC in New Jersey is explored, from how the response was coordinated to how it ultimately was executed. The state coordinated its response on multiple levels and, as such, tested the practical applicability of the EMAC process and employed best practices and solutions to issues that arose. These best practices and solutions may prove invaluable for any state or territory that may activate the EMAC system for emergency medical service resources.

  14. Back strength and flexibility of EMS providers in practicing prehospital providers.

    PubMed

    Crill, Matthew T; Hostler, David

    2005-06-01

    In the execution of prehospital care duties, an EMS provider may be required to carry equipment and patients over long distances or over multiple flights of stairs at any time of the day. At a minimum, a prehospital provider must have sufficient lower back strength and hamstring flexibility to prevent musculoskeletal injury while lifting. This study administered fitness assessments related to the occupational activities of the prehospital provider with the purpose of describing the incidence of occupational back injury and percentage of providers with known risk factors for back injury. Ninety subjects were tested during a regional EMS conference. Men were significantly taller and heavier than women and had significantly less hamstring flexibility. Body Mass Index was 30.7 +/- 7.2 in men and 28 +/- 5.7 in women. However, no significant differences were noted in an extension test of back strength. When surveyed, 47.8% of subjects reported a back injury in the previous 6 months but only 39.1% of these injuries were sustained while performing EMS duties. While only 13% of these injuries resulted in missed work, 52.2% reported their injury interfered with their daily activities. In spite of the physical nature of the profession, EMS providers in our sample were significantly overweight according to their Body Mass Index and may lack sufficient back strength and flexibilityfor safe execution of their duties. This group of professionals may be at risk for occupational injury and should be targeted for interventions to improve strength and flexibility.

  15. An EM Algorithm for Maximum Likelihood Estimation of Process Factor Analysis Models

    ERIC Educational Resources Information Center

    Lee, Taehun

    2010-01-01

    In this dissertation, an Expectation-Maximization (EM) algorithm is developed and implemented to obtain maximum likelihood estimates of the parameters and the associated standard error estimates characterizing temporal flows for the latent variable time series following stationary vector ARMA processes, as well as the parameters defining the…

  16. Saudi EMS Students' Perception of and Attitudes toward Their Preparedness for Disaster Management

    ERIC Educational Resources Information Center

    Alrazeeni, Daifallah

    2015-01-01

    Background: Disasters led not only to the loss of life and destruction of public infrastructures, but also resulted in consequent healthcare delivery concerns. Disaster preparedness is considered one of the key steps in emergency management. EMS students had very scanty knowledge, attitude and practices about disaster preparedness and mitigation.…

  17. Are you under stress in EMS. Understanding the slippery slope of burnout and PTSD.

    PubMed

    Collopy, Kevin T; Kivlehan, Sean M; Snyder, Scott R

    2012-10-01

    Burnout and PTSD are closely linked and often underreported in EMS. EMS classrooms do little or nothing to prepare providers for the inherent emotional stresses of emergency response and the "thick skin" culture of EMS may make many providers apprehensive about sharing their true feelings. Burnout is triggered by many of the same stresses that lead to the symptoms of PTSD and providers experiencing burnout that doesn't resolve within a few weeks may actually be experiencing PTSD. Be mindful of yourself and your fellow coworkers, particularly after a very traumatic response. And remember traumatic responses don't need to be as dramatic as Sept. 11, New Orleans after Hurricane Katrina or the Aurora, CO shootings to bother an EMS worker. In contrast, these are the calls where providers often receive the most attention. Instead, watch for the new father who just performed CPR on an infant the same age as his own, or the provider who just watched his or her friend die following a motor vehicle collision. Pay attention to yourself and colleagues, and be responsible and honest with yourself and others about when coping strategies are enough, and when they aren't. Finally, don't ever be afraid to seek help. PMID:23097838

  18. The Role of Journals in Building up Communities: The Experience of "Ciência em Tela"

    ERIC Educational Resources Information Center

    Martins, Isabel; de Souza Barros, Susana

    2015-01-01

    "Ciência em Tela" is an open access Brazilian science teacher education online journal that has been published twice a year since 2008 and which encourages the participation of professionals with different backgrounds and the submission of a variety of textual genres, besides research papers. Another feature is that the journal includes…

  19. Injury risks of EMS responders: evidence from the National Fire Fighter Near-Miss Reporting System

    PubMed Central

    Taylor, Jennifer A; Davis, Andrea L; Barnes, Brittany; Lacovara, Alicia V; Patel, Reema

    2015-01-01

    Objectives We analysed near-miss and injury events reported to the National Fire Fighter Near-Miss Reporting System (NFFNMRS) to investigate the workplace hazards and safety concerns of Emergency Medical Services (EMS) responders in the USA. Methods We reviewed 769 ‘non-fire emergency event’ reports from the NFFNMRS using a mixed methods approach. We identified 185 emergency medical calls and analysed their narrative text fields. We assigned Mechanism of Near-Miss/Injury and Nature of Injury codes and then tabulated frequencies (quantitative). We coded major themes regarding work hazards and safety concerns reported by the EMS responders (qualitative). Results Of the 185 emergency medical calls, the most commonly identified Mechanisms of Near-Miss/Injury to EMS responders was Assaults, followed by Struck-by Motor Vehicle, and Motor Vehicle Collision. The most commonly identified weapon used in an assault was a firearm. We identified 5 major domains of workplace hazards and safety concerns: Assaults by Patients, Risks from Motor Vehicles, Personal Protective Equipment, Relationships between Emergency Responders, and Policies, Procedures and Practices. Conclusions Narrative text from the NFFNMRS is a rich source of data that can be analysed quantitatively and qualitatively to provide insight into near-misses and injuries sustained by EMS responders. Near-miss reporting systems are critical components for occupational hazard surveillance. PMID:26068510

  20. DeepPicker: A deep learning approach for fully automated particle picking in cryo-EM.

    PubMed

    Wang, Feng; Gong, Huichao; Liu, Gaochao; Li, Meijing; Yan, Chuangye; Xia, Tian; Li, Xueming; Zeng, Jianyang

    2016-09-01

    Particle picking is a time-consuming step in single-particle analysis and often requires significant interventions from users, which has become a bottleneck for future automated electron cryo-microscopy (cryo-EM). Here we report a deep learning framework, called DeepPicker, to address this problem and fill the current gaps toward a fully automated cryo-EM pipeline. DeepPicker employs a novel cross-molecule training strategy to capture common features of particles from previously-analyzed micrographs, and thus does not require any human intervention during particle picking. Tests on the recently-published cryo-EM data of three complexes have demonstrated that our deep learning based scheme can successfully accomplish the human-level particle picking process and identify a sufficient number of particles that are comparable to those picked manually by human experts. These results indicate that DeepPicker can provide a practically useful tool to significantly reduce the time and manual effort spent in single-particle analysis and thus greatly facilitate high-resolution cryo-EM structure determination. DeepPicker is released as an open-source program, which can be downloaded from https://github.com/nejyeah/DeepPicker-python. PMID:27424268