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In vitro and in vivo application of RNA interference for targeting genes involved in peritrophic matrix synthesis in a lepidopteran system

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Insect Science 2013年第1期20卷 92-100页

作者： Umut Toprak Doug Baldwin Martin Erlandson Cedric Gillott Stephanie Harris dwayne D. hegedus Agriculture and Agri-Food Canada Ottawa Ontario Canada Department of Biology University of Saskatchewan Canada Department of Plant Protection Faculty of Agriculture University of Ankara Turkey Department of Food and Bioproduct Sciences University of Saskatchewan Saskatoon SK Canada

The midgut of most insects is lined with a semipermeable acellular tube, the peritrophic matrix （PM）, composed of chitin and proteins. Although various genes encoding PM proteins have been characterized, our understanding of their roles in PM structure and function is very limited. One promising approach for obtaining functional information is RNA interference, which has been used to reduce the levels of specific mRNAs using double-stranded RNAs administered to larvae by either injection or feeding. Although this method is well documented in dipterans and coleopterans, reports of its success in lepidopterans are varied. In the current study, the silencing midgut genes encoding PM proteins （insect intestinal mucin 1, insect intestinal mucin 4, PM protein 1） and the chitin biosynthetic or modifying enzymes （chitin synthase-B and chitin deacetylase 1） in a noctuid lepidopteran, Mamestra configurata, was examined in vitro and in vivo. In vitro studies in primary midgut epithelial cell preparations revealed an acute and rapid silencing （by 24 h） for the gene encoding chitin deacetylase 1 and a slower rate of silencing （by 72 h） for the gene encoding PM protein 1. Genes encoding insect intestinal mucins were slightly silenced by 72 h, whereas no silencing was detected for the gene encoding chitin synthase-B. In vivo experiments focused on chitin deacetylase 1, as the gene was silenced to the greatest extent in vitro. Continuous feeding of neonates and fourth instar larvae with double-stranded RNA resulted in silencing of chitin deacetylase 1 by 24 and 36 h, respectively. Feeding a single dose to neonates also resulted in silencing by 24 h. The current study demonstrates that genes encoding PM proteins can be silenced and outlines conditions for RNA interference by per os feeding in lepidopterans.

关键词： chitin deacetylase chitin synthase-B lepidoptera peritrophic matrix,peritrophin RNA interference

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Proteomics analysis of Trichoplusia ni midgut epithelial cell brush border membrane vesicles

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Insect Science 2019年第3期26卷 424-440页

作者： Muhammad Afzal Javed Cathy Coutu David A. Theilmann Martin A. Erlandson dwayne D. hegedus Saskatoon Research and Development Centre Agriculture and Agri-Food Canada Saskatoon Saskatchewan Canada Summerland Research and Development Centre Agriculture and Agri-Food Canada Summerland British Columbia Canada Department of Food & Bio-Product Sciences University of Saskatchewan Saskatoon Saskatchewan Canada

The insect midgut epithelium is composed of columnar, goblet, and regenerative cells. Columnar epithelial cells are the most abundant and have membrane protrusions that form the brush border membrane (BBM) on their apical side. These increase surface area available for the transport of nutrients, but also provide opportunities for interaction with xenobiotics such as pathogens, toxins and host plant allelochemicals. Recent improvements in proteomic and bioinfbrmatics tools provided an opportunity to determine the proteome of the T. ni BBM in unprecedented detail. This study reports the identification of proteins from BBM vesicles (BBMVs) using single dimension polyacrylamide gel elec? trophoresis coupled with multi-dimensional protein identification technology. More than 3000 proteins were associated with the BBMV of which 697 were predicted to possess either a signal peptide, at least one transmembrane domain or a GPI-anchor signal. Of these, bioinfbrmatics analysis and manual curation predicted that 185 may be associated with the BBMV or epithelial cell plasma membrane. These are discussed with respect to their predicted functions, namely digestion, nutrient uptake, cell signaling, development, cell-cell interactions, and other functions. We believe this to be the most detailed proteomic analysis of the lepidopteran midgut epithelium membrane to date, which will provide information to better understand the biochemical, physiological and pathological processes taking place in the larval midgut.

关键词： brush border membrane vesicles midgut proteomic analysis Trichoplusiani

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Identification of the Mamestra configurata （Lepidoptera Noctuidae） peritrophic matrix proteins and enzymes involved in peritrophic matrix chitin metabolism

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Insect Science 2016年第5期23卷 656-674页

作者： Umut Toprak Martin Erlandson Doug Baldwin Steve Karcz Lianglu Wan Cathy Coutu Cedric Gillott dwayne D. hegedus Agriculture and Agri-Food Canada Saskatoon SK Canada Department of Biology University of Saskatchewan Saskatoon SK Canada Department of Plant Protection Faculty of Agriculture University of Ankara Ankara Turkey Department of Food and Bioproduct Sciences University of Saskatchewan Saskatoon SK Canada plant Biotechnology Institute National Research Council of Canada Saskatoon SK Canada

The peritrophic matrix （PM） is essential for insect digestive system physiol- ogy as it protects the midgut epithelium from damage by food particles, pathogens, and toxins. The PM is also an attractive target for development of new pest control strategies due to its per os accessibility. To understand how the PM performs these functions, the molecular architecture of the PM was examined using genomic and proteomic approaches in Mamestra configurata （Lepidoptera： Noctuidae）, a major pest of cruciferous oilseed crops in North America. Liquid chromatography-tandem mass spectrometry analyses of the PM identified 82 proteins classified as：（i） peritrophins, including a new class with a CBDIII domain; （ii） enzymes involved in chitin modification （chitin deacetylases）, di- gestion （serine proteases, aminopeptidases, carboxypeptidases, lipases and ^-amylase） or other reactions （/^-l,3-glucanase, alkaline phosphatase, dsRNase, astacin, pantetheinase）; （iii） a heterogenous group consisting of polycalin, REPATs, serpin, C-Type lectin and Lsti99/Lsti201 and 3 novel proteins without known orthologs. The genes encoding PM proteins were expressed predominantly in the midgut, cDNAs encoding chitin synthase-2 （McCHS-2）, chitinase （McCHI）, and fl-N-acetylglucosaminidase （McNAG） enzymes, in- volved in PM chitin metabolism, were also identified. McCHS-2 expression was specific to the midgut indicating that it is responsible for chitin synthesis in the PM, the only chitinous material in the midgut. In contrast, the genes encoding the chitinolytic enzymes were expressed in multiple tissues. McCHS-2, McCHI, and McNAG were expressed in the midgut of feeding larvae, and NAG activity was present in the PM. This information was used to generate an updated model of the lepidopteran PM architecture.

关键词： chitin Mamestra configurata peritrophic matrix protein

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Evaluation of a <i>Brassica napus</i>Auxin-Repressed Gene Induced by Flea Beetle Damage and <i>Sclerotinia sclerotiorum</i>Infection

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American Journal of Plant Sciences 2017年第8期8卷 1921-1952页

作者： Limin Wu Min Yu Jennifer Holowachuk Andrew Sharpe Derek Lydiate dwayne hegedus Margaret Gruber InnoTech Alberta Vegreville Canada Saskatoon Research Centre Agriculture and Agri-Food Canada Saskatoon Canada Global Institute for Food Security University of Saskatchewan Saskatoon Canada

Biotic stresses negatively affect canola growth and production. Flea beetle damage and Sclerotinia sclerotiorum (S. sclerotiorum) infection are two of the worst biotic stresses for canola. Auxin Repressed Proteins (ARPs) responsive to several abiotic stresses have been reported. However, information about ARPs induced by Flea beetle damage and S. sclerotiorum infection, their roles in biotic stress tolerance are still lacking in canola. ESTs for an Auxin Repressed Protein 1 (BnARP1) were highly represented (expressed) in a Brassica napus subtractive library developed after leaf damage by the crucifer flea beetle (Phyllotreta cruciferae). Expression of this gene was under different developmental control in B. napus, and it was co-induced in B. napus by flea beetle feeding, S. sclerotiorum infection, drought and cold. A total of 25 BnARP genes were represented in different B. napus stress and development EST libraries and indicated larger, diversified families than known earlier. Dwarf phenotypes, primary root growth inhibition, lateral root enhancement, reduced sensitivity to 2, 4-D, and reduced PIN1 and LOX expression in transgenic Arabidopsis expression lines suggest that BnARP1 is an auxin repressor that prevents auxin transport and supports an interaction between the auxin and jasmonate signalling pathways. And the increased survival after S. sclerotiorum infection in transgenic over-expression Arabidopsis suggests that BnARP1 could play a role in S. sclerotiorum tolerance through connecting auxin and jasmonate signalling pathways.

关键词： BnARP Brassica napus Flea Beetle S. sclerotiorum

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Soybean Response to Weed Residues in the Soil

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Agricultural Sciences 2024年第8期15卷 801-811页

作者： dwayne D. Joseph Michael W. Marshall Matthew Cutulle Department of Plant and Environmental Sciences Clemson University Clemson SC USA Edisto Research and Education Center Clemson University Blackville SC USA Coastal Research and Education Center Clemson University Charleson SC USA

Soybean production systems that return plant residues to the soil surface are gaining in popularity. As these practices become more widespread, more crop and weed residues are being introduced into the upper soil profile. Greenhouse studies were conducted to determine the effects of varying concentrations of Palmer amaranth and pitted morningglory plant residues (aboveground portion of the plant) on soybean production. The study was arranged in a completely randomized experimental design with five treatments and five replications. Palmer amaranth and pitted morningglory residues were incorporated into soil at 20,000, 40,000, 80,000 and 160,000 ppm. Inert plastic residue at the same residue levels was included as a check. Soybean dry weight, leaf area and leaf tissue nutrient content were recorded during the study. A decrease in soybean dry weight and leaf area was observed as Palmer amaranth residue in the soil increased. Palmer amaranth residues of 160,000 ppm and 80,000 ppm in the soil significantly reduced soybean dry weight by 69% and 59%, respectively, and soybean leaf area by 60% and 57%, respectively. In contrast, pitted morningglory and inert plastic residues had no observable effect on soybean growth and development. This study demonstrated Palmer amaranth residues in the soil impacted early season soybean growth and development.

关键词： Reduced Tillage Plant Residues Allelopathy Glycine max L.

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The effects of a daily, 6-week exergaming curriculum on balance in fourth grade children

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Journal of Sport and Health Science 2013年第3期2卷 131-137页

作者： dwayne P.Sheehan Larry Katz Department of Physical Education and Recreation Studies Mount Royal University Faculty of Kinesiology University of Calgary

Balance is an essential component of movement and is critical in the ability to participate in physical activity. Developing an exergaming curriculum for schools has the potential to improve balance or postural stability in children. In this study, a purposely-built exergaming center in an elementary school was used to test fourth grade students with a specially designed exergaming curriculum oriented toward improving postural stability. The program was implemented over a 6-week period, 34 min per day, 4-5 days per week. Two control groups were used：（1） a physical education （PE） class geared toward agility, balance, and coordination （ABC） improvement, and （2） a typical PE curriculum class. Exergaming students improved their postural stability significantly over a 6-week period compared to those in the typical PE class. Improvements in postural stability were also evident in the ABC class. Postural stability in the girls was better than the boys in all pre- and post-intervention tests. This study demonstrates that exergaming is a practical resource in the PE class to improve postural stability.

关键词： Balance Children Elementary school Exergaming Fundamental movement skills

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Centrifuge and numerical modeling of h-type anti-slide pile reinforced soil-rock mixture slope

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Journal of Mountain Science 2023年第5期20卷 1441-1457页

作者： ZHANG Hao XING Hao-feng XUE Dao-rui TANNANT dwayne Department of Geotechnical Engineering College of Civil EngineeringTongji UniversityShanghai 200092China School of Urban Construction and Transportation Hefei UniversityHefei 230601China School of Engineering University of British ColumbiaKelowna BC V1V 1V7Canada Guangdong Communication Planning and Design Institute Group Co. Ltd.Guangzhou 510507China

Due to the loose structure,high porosity and high permeability of soil-rock mixture slope,the slope is unstable and may cause huge economic losses and *** h-type anti-slide pile is regarded as an effective means to prevent the instability of soilrock mixture *** this paper,a centrifuge model test was conducted to investigate the stress distribution of the h-type anti-slide pile and the evolution process of soil arching during the loading.A numerical simulation model was built based on the similar relationship between the centrifuge model and the prototype to investigate the influence factors of the pile spacing,anchored depth,and crossbeam stiffness,and some recommendations were proposed for its *** results show that the bending moment distribution of the rear pile exhibits Wshaped,while for the front pile,its distribution resembles *** soil arching evolution process during loading is gradually dissipated from bottom to top and from far to *** the loading,the change of bending moment can be divided into three stages,namely,the stabilization stage,the slow growth stage,and the rapid growth *** engineering projects,the recommended values of the pile spacing,anchored depth,and crossbeam stiffness are 4.0d,2.0d,and 2.0EI,where d and EI are the diameter and bending stiffness of the h-type anti-slide pile respectively.

关键词： Centrifugemodel test Numerical simulation h-type anti-slide pile Soil-rock mixture slop Soil arching

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A multi-step approach for block-cave production scheduling optimization

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International Journal of Mining Science and Technology 2013年第5期23卷 739-750页

作者： Yashar Pourrahimian Hooman Askari-Nasab dwayne D.Tannant Department of Civil and Environmental Engineering School of Mining and Petroleum EngineeringUniversity of Alberta School of Engineering University of British Columbia

Production schedules that provide optimal operating strategies while meeting practical,technical,and environmental constraints are an inseparable part of mining *** only on manual planning methods or computer software based on heuristic algorithms will lead to mine schedules that are not the optimal global *** mine planning models have been proved to be very effective in supporting decisions on sequencing the extraction of material in *** objective of this paper is to develop a practical optimization framework for caving operations’production *** overcome the size problem of mathematical programming models and to generate a robust practical near-optimal schedule,a multi-step method for long-term production scheduling of block caving is presented.A mixed-integer linear programming(MILP)formulation is used for each *** formulations are developed,implemented,and verifed in the TOMLAB/CPLEX *** production scheduler aims to maximize the net present value of the mining operation while the mine planner has control over defned *** and comparison of the models for production scheduling using 298 drawpoints over 15 periods are presented.

关键词： Block caving Aggregation Production scheduling MILP CPLEX

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Surface texture and friction characteristics of diamond-ground concrete and asphalt pavements

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Journal of Traffic and Transportation Engineering(English Edition) 2016年第5期3卷 475-482页

作者： Shuo Li dwayne Harris Tim Wells Division of Research and Development Indiana Department of Transportation

Recently, diamond grinding has gained increasing attention as pavement preservation treatment to restore desired surface characteristics, particularly friction. Compared to other pavement preservation treatments such as surface overlays and high friction surface treat- ments, diamond grinding may cost less, save construction time, or require minimum maintenance. Diamond grinding produces longitudinal, continuous, and line-type texture that contains corrugations with evenly spaced ridges. The improved surface texture will immediately enhance pavement surface friction and reduce the possibility of hydroplaning in rainy weather. However, little information has been documented on the texture charac- teristics and long-term friction performance of diamond-ground pavements. A field evalu- ation was conducted to examine the surface texture and friction characteristics in diamond- ground concrete and asphalt pavements by the authors. Five pavement test sections, including two diamond-ground concrete pavements, one diamond-ground concrete bridge deck, one diamond-ground asphalt pavement, and one transversely tined concrete pave- ment, were selected for evaluation. Laser scanner testing was performed to capture both macro and microtexture profiles. Locked wheel testing was performed to measure the fric- tion numbers. The test results were examined and compared so as to evaluate two performances, after construction and long-term friction performance of diamond-ground pavements. It was found that longitudinal diamond grinding can provide durable, satisfactory surface friction performance for both concrete and asphalt pavements.

关键词： Concrete pavement Asphalt pavement Surface friction Diamond-grinding Macrotexture Microtexture

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Roles for the VCP co-factors Npl4 and Ufd1 in neuronal function in Drosophila melanogaster

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Journal of Genetics and Genomics 2017年第10期44卷 493-501页

作者： dwayne J.Byrne Mark J.Harmon Jeremy C.Simpson Craig Blackstone Niamh C. O'Sullivan UCD School of Biomolecular and Biomedical Science UCD Conway Institute University College Dublin Dublin 4 Ireland UCD School of Biology and Environmental Science UCD Conway Institute of Biomolecular and Biomedical Research University College Dublin Dublin 4Ireland Cell Biology Section Neurogenetics Branch National Institute of Neurological Disorders and Stroke National Institutes of Health Bethesda MD 20892 USA

The VCP-Ufd1-Npl4 complex regulates proteasomal processing within cells by delivering ubiquitinated proteins to the proteasome for *** in VCP are associated with two neurodegenerative diseases,amyotrophic lateral sclerosis（ALS） and inclusion body myopathy with Paget＇s disease of the bone and frontotemporal dementia（IBMPFD）,and extensive study has revealed crucial functions of VCP within *** contrast,little is known about the functions of Npl4 or Ufd1 in *** neuronalspecific knockdown of Npl4 or Ufd1 in Drosophila melanogaster,we infer that Npl4 contributes to microtubule organization within developing motor ***,Npl4 RNAi flies present with neurodegenerative phenotypes including progressive locomotor deficits,reduced lifespan and increased accumulation of TAR DNA-binding protein-43 homolog（TBPH）.Knockdown,but not overexpression,of TBPH also exacerbates Npl4 RNAi-associated adult-onset neurodegenerative *** contrast,we find that neuronal knockdown of Ufd1 has little effect on neuromuscular junction（NMJ） organization,TBPH accumulation or adult *** findings suggest the differing neuronal functions of Npl4 and Ufd1 in vivo.

关键词： Proteasome TDP-43 accumulation Neurodegeneration Drosophila

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