Proteomics of pupal brains in Sarcophaga crassipalpis: first database of diapause-associated proteins

Most molecular work on insect diapause has focused on the expression of unique diapause transcripts, rather than the protein products. Here we present our first results from a proteomic comparison of diapausing and nondiapausing pupal brains. Proteins extracted from diapausing pupal brains in the flesh fly Sarcophaga crassipalpis were separated by two-dimensional gel electrophoresis and compared with those from nondiapausing pupal brains. Unique proteins and proteins expressed at different levels in diapausing and nondiapausing brains were identified by Nano-LC/MS/MS (capillary-liquid chromatography-nanospray tandem mass spectrometry). With this approach and Coomassie staining we detected 17 unique or upregulated (≥3x) spots, and 16 spots that were missing or downregulated in diapause. Most of the brain proteins present in higher amounts during diapause were heat shock proteins (members of the HSP70 and small HSP families). Brain proteins that were less abundant in diapause include phosphoenolpyruvate synthase, fatty acid binding protein, endonuclease, retinal pigment epithelium 65-protein,Ylt1, 16S rRNA pseudouridylate synthase, pupative S-transferase, and EG0003.7. Our 2-D proteome maps include many additional unknown proteins. While the mRNAs encoding certain of these proteins (e.g.HSPs) were previously known to be associated with diapause, many of the other proteins were not known to be linked to diapause, thus suggesting that the proteomic approach nicely supplements work done at the transcript level.NSF grant IOB-041672

Effects of in vitro fertilization and embryo culture on TRP53 and Bax expression in B6 mouse embryos

In the mouse, embryo culture results in a characteristic phenotype of retarded embryo preimplantation development and reduced numbers of cells within embryos. The expression of TRP53 is central to the regulation of the cell's capacity to proliferate and survive. In this study we found that Trp53 mRNA is expressed throughout the preimplantation stage of development. Levels of TRP53 protein expression were low during the cleavage stages and increased at the morula and blastocyst stages in B6 embryos collected from the reproductive tract. Embryos collected at the zygote stage and cultured for 96 h also showed low levels of TRP53 expression at precompaction stages. There were higher levels of TRP53 in cultured morula and the level in cultured blastocysts was clearly increased above blastocysts collected directly from the uterus. Immunolocalization of TRP53 showed that its increased expression in cultured blastocysts corresponded with a marked accumulation of TRP53 within the nuclei of embryonic cells. This pattern of expression was enhanced in embryos produced by in vitro fertilization and subjected to culture. The TRP53 was transcriptionally active since culture also induced increased expression of Bax, yet this did not occur in embryos lacking Trp53 (Trp53-/-). The rate of development of Trp53-/- zygotes to the blastocyst stage was not different to wildtype controls when embryos were cultured in groups of ten but was significantly faster when cultured individually. The results show that zygote culture resulted in the accumulation of transcription activity of TRP53 in the resulting blastocysts. This accounts for the adverse effects of culture of embryos individually, but does not appear to be the sole cause of the retarded preimplantation stage growth phenotype associated with culture in vitro

Vehicle Communication using Secrecy Capacity

We address secure vehicle communication using secrecy capacity. In particular, we research the relationship between secrecy capacity and various types of parameters that determine secrecy capacity in the vehicular wireless network. For example, we examine the relationship between vehicle speed and secrecy capacity, the relationship between the response time and secrecy capacity of an autonomous vehicle, and the relationship between transmission power and secrecy capacity. In particular, the autonomous vehicle has set the system modeling on the assumption that the speed of the vehicle is related to the safety distance. We propose new vehicle communication to maintain a certain level of secrecy capacity according to various parameters. As a result, we can expect safer communication security of autonomous vehicles in 5G communications.Comment: 17 Pages, 12 Figure

Thermal Management For Efficient Fast Refill Of Compressed Natural Gas

Paper presented at 2018 Canadian Society of Mechanical Engineers International Congress, 27-30 May 2018.As an alternative fuel, compressed natural gas (CNG) plays an important role fueling transportation. Like driving other types of vehicles, drivers of CNG-powered vehicles prefer to refill on-board CNG tanks in a short time, which is referred to as fast refill. Accompanied by a fast pressure increase in the cylinder, temperature will also rise significantly due to re-compression work with limited time for the heat to dissipate. This phenomenon ends with a common problem referred to as under-refill, which means less mass filled than the standards. This article presents a summary of the research works to solve this problem and improve fill efficiency by thermal management. Twodimensional axisymmetric CFD (computational fluid dynamics) simulations of unsteady, compressible, turbulent flow in fast refill process are conducted. Three different types of thermal management concepts are discussed, including active cooling, pre-chilling and real-time chilling. The results show that thermal management is a promising way to mitigate the under-refill problem with a maximum of 7% increase of fill efficiency achieved by active cooling. These provide guidelines for future research on improving fill efficiency through cooling strategies in which real-time chilling should be firstly focused on

Targeted Ablation of Distal Cerebrospinal Fluid-Contacting Nucleus Alleviates Renal Fibrosis in Chronic Kidney Disease

The potential function of distal cerebrospinal fluid-contacting nucleus (dCSF-CNs) in chronic kidney disease (CKD) development is poorly understood. We hypothesized that dCSF-CNs might affect the renin-angiotensin system (RAS) in kidney injury progression, with dCSF-CNs ablation potentially alleviating local RAS and renal fibrosis in rats after five-sixths nephrectomy (5/6Nx). Part of rats were randomly administered artificial cerebrospinal fluid (aCSF) intracerebroventricularly (icv), followed by 5/6Nx or sham operation; and other part of rats were administered Cholera toxin B subunit conjugated with saporin (CB-SAP) for dCSF-CNs lesion before 5/6Nx. The effect of CB-SAP on dCSF-CNs ablation was confirmed by double immunofluorescence staining. RAS component, NOX2 and c-fos levels in the subfornical organ (SFO), hypothalamic paraventricular nucleus (PVN) and hippocampus, as well as tyrosine hydroxylase (TH) and c-fos positive cells in rostral ventrolateral medulla (RVLM) were assessed. Next, the levels of RAS components (angiotensinogen [AGT], angiotensin-converting enzyme [ACE], Ang II type 1 receptor [AT1R], angiotensin-converting enzyme 2 [ACE2], and Mas receptor), NADPH oxidases (NOX2 and catalase), inflammatory cytokines (monocyte chemotactic protein 1 [MCP-1] and IL-6), and fibrotic factors (fibronectin and collagen I) were assessed. Less CB-labeled neurons were found in dCSF-CNs of CB-SAP-treated rats compared with 5/6Nx animals. Meanwhile, CB-SAP downregulated AGT, Ang II, AT1R, NOX2, catalase, MCP-1, IL-6, fibronectin, and collagen I, and upregulated ACE2 and Mas receptor, compared with CKD rats. More TH and c-fos positive cells were found in RVLM of 5/6Nx rats but the number decreased after dCSF-CNs ablation. Targeted dCSF-CNs ablation could alleviate renal inflammation and fibrosis in chronic kidney injury by inhibiting cerebral and renal RAS/NADPH oxidase

Preparation of Black Copper Electrolyte by Acid Oxidation Leaching of Refining Slag

This is an article in the field of metallurgical engineering. The preparation of black copper electrolyte by removing copper from secondary refining slag of high copper and high tin anode furnace by hydrogen peroxide oxidation and acid leaching was studied. The process conditions related to the removal efficiency of the copper and arsenic were researched including the hydrogen peroxide dosage, reaction temperature, sulfuric acid concentration and reaction time. Results show that, under the following optimal conditions, including refining slag of 20.0 g, 2.5 mol/L sulfuric acid of 200 mL, solid-liquid ratio of 1/10, temperature of 65 ℃, dosage of hydrogen peroxide of 10.0 mL, stirring speed of 400 r/min and leaching time of 60 min, the leaching ratios of Cu and As can reach 88.98% and 87.33%, respectively, and the leaching ratios of Ni and Sn were only 7.72% and 1.34%, respectively. The concentration of copper ion in the leaching solution was 48.48 g/L, which can be used as the supplement of black copper electrolyte

Exploring the shared genes of hypertension, diabetes and hyperlipidemia based on microarray

Given their relationship with metabolic syndrome and systematic inflammatory diseases, the pathogenesis of hypertension, hyperglycemia, and hyperlipidemia is closely related. To explore the common genes among these three conditions, spontaneous hypertensive rats (SHR), spontaneous diabetic Goto-Kakizaki rats (GK) and hyperlipidemia rats (HMR) were reared for experiments. Gene array was used to identify the genes of SHR, GK and HMR compared with normal Wistar rats using TBtools software. First, real-time PCR was applied to verify these genes, and Cytoscape software was used to construct networks based on the National Center for Biotechnology Information (NCBI) database. Second, Kyoto Encyclopedia of Genes and Genomes (KEGG) database analysis was performed to classify the genes. Visualization and Integrated Discovery (DAVID) database and Gene Ontology database were used to explore the biological function. Finally, Onto-tools Pathway Express was used to analyze the pathways of shared genes. Importantly, upregulated common genes, such as Bad, Orm1, Arntl and Zbtb7a, were used to construct a network of 150 genes, while downregulated genes, such as Mif and Gpx1, formed a network of 29 genes. Interestingly, the networks were involved in various pathways, such as insulin signal pathway, endometrial cancer pathway, circadian rhythm pathway, and pancreatic cancer pathway. We discovered common genes of SHR, GK and HMR compared with normal Wistar rats, and the association of these genes together with biological function were preliminarily revealed

Integrated transcriptomics and metabolomics analysis provide insight into the resistance response of rice against brown planthopper

IntroductionThe brown planthopper (Nilaparvata lugens Stål, BPH) is one of the most economically significant pests of rice. The Bph30 gene has been successfully cloned and conferred rice with broad-spectrum resistance to BPH. However, the molecular mechanisms by which Bph30 enhances resistance to BPH remain poorly understood.MethodsHere, we conducted a transcriptomic and metabolomic analysis of Bph30-transgenic (BPH30T) and BPH-susceptible Nipponbare plants to elucidate the response of Bph30 to BPH infestation.ResultsTranscriptomic analyses revealed that the pathway of plant hormone signal transduction enriched exclusively in Nipponbare, and the greatest number of differentially expressed genes (DEGs) were involved in indole 3-acetic acid (IAA) signal transduction. Analysis of differentially accumulated metabolites (DAMs) revealed that DAMs involved in the amino acids and derivatives category were down-regulated in BPH30T plants following BPH feeding, and the great majority of DAMs in flavonoids category displayed the trend of increasing in BPH30T plants; the opposite pattern was observed in Nipponbare plants. Combined transcriptomics and metabolomics analysis revealed that the pathways of amino acids biosynthesis, plant hormone signal transduction, phenylpropanoid biosynthesis and flavonoid biosynthesis were enriched. The content of IAA significantly decreased in BPH30T plants following BPH feeding, and the content of IAA remained unchanged in Nipponbare. The exogenous application of IAA weakened the BPH resistance conferred by Bph30.DiscussionOur results indicated that Bph30 might coordinate the movement of primary and secondary metabolites and hormones in plants via the shikimate pathway to enhance the resistance of rice to BPH. Our results have important reference significance for the resistance mechanisms analysis and the efficient utilization of major BPH-resistance genes

Comprehensive identification and characterization of lncRNAs and circRNAs reveal potential brown planthopper-responsive ceRNA networks in rice

Brown planthopper (Nilaparvata lugens Stål, BPH) is one of the most destructive pests of rice. Non-coding RNA plays an important regulatory role in various biological processes. However, comprehensive identification and characterization of long non-coding RNAs (lncRNAs) and circular RNAs (circRNAs) in BPH-infested rice have not been performed. Here, we performed a genome-wide analysis of lncRNAs and circRNAs in BPH6-transgenic (resistant, BPH6G) and Nipponbare (susceptible, NIP) rice plants before and after BPH feeding (early and late stage) via deep RNA-sequencing. A total of 310 lncRNAs and 129 circRNAs were found to be differentially expressed. To reveal the different responses of resistant and susceptible rice to BPH herbivory, the potential functions of these lncRNAs and circRNAs as competitive endogenous RNAs (ceRNAs) were predicted and investigated using Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses. Dual-luciferase reporter assays revealed that miR1846c and miR530 were targeted by the lncRNAs XLOC_042442 and XLOC_028297, respectively. In responsive to BPH infestation, 39 lncRNAs and 21 circRNAs were predicted to combine with 133 common miRNAs and compete for miRNA binding sites with 834 mRNAs. These mRNAs predictably participated in cell wall organization or biogenesis, developmental growth, single-organism cellular process, and the response to stress. This study comprehensively identified and characterized lncRNAs and circRNAs, and integrated their potential ceRNA functions, to reveal the rice BPH-resistance network. These results lay a foundation for further study on the functions of lncRNAs and circRNAs in the rice-BPH interaction, and enriched our understanding of the BPH-resistance response in rice

Untargeted metabolomics of the cochleae from two laryngeally echolocating bats

High-frequency hearing is regarded as one of the most functionally important traits in laryngeally echolocating bats. Abundant candidate hearing-related genes have been identified to be the important genetic bases underlying high-frequency hearing for laryngeally echolocating bats, however, extensive metabolites presented in the cochleae have not been studied. In this study, we identified 4,717 annotated metabolites in the cochleae of two typical laryngeally echolocating bats using the liquid chromatography–mass spectroscopy technology, metabolites classified as amino acids, peptides, and fatty acid esters were identified as the most abundant in the cochleae of these two echolocating bat species, Rhinolophus sinicus and Vespertilio sinensis. Furthermore, 357 metabolites were identified as significant differentially accumulated (adjusted p-value <0.05) in the cochleae of these two bat species with distinct echolocating dominant frequencies. Downstream KEGG enrichment analyses indicated that multiple biological processes, including signaling pathways, nervous system, and metabolic process, were putatively different in the cochleae of R. sinicus and V. sinensis. For the first time, this study investigated the extensive metabolites and associated biological pathways in the cochleae of two laryngeal echolocating bats and expanded our knowledge of the metabolic molecular bases underlying high-frequency hearing in the cochleae of echolocating bats