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https://www.readbyqxmd.com/read/29776310/preparation-and-evaluation-of-ribonuclease-resistant-viral-hiv-rna-standards-based-on-armored-rna-technology
#1
Mohammad Gholami, Mehrdad Ravanshad, Kazem Baesi, Siamak M. Samiee, Negin Hosseini Rozbahani, Minoo Mohraz
BAckground: The human immunodeficiency virus type 1 (HIV-1) is an infectious viral agent that gradually extinguishes the immune system, resulting in acquired immune deficiency syndrome (AIDS). The aim of this study was to construct an RNA-positive control based on armored (AR) RNA technology, using HIV-1 RNA as a model. Methods: The MS2 maturase, a coat protein gene (at positions 1765 to 1787) and HIV-1 pol gene were cloned into pET-32a plasmid. The prepared plasmid was transformed into Escherichia coli strain BL2 (DE3), and the expression of the construct was induced by 1 mM of isopropyl-L-thio-D-galactopyranoside (IPTG) at 37 °C for 16 h to obtain the fabricated AR RNA...
May 19, 2018: Iranian Biomedical Journal
https://www.readbyqxmd.com/read/29755489/optimizing-production-in-the-new-generation-of-apricot-cultivars-self-incompatibility-s-rnase-allele-identification-and-incompatibility-group-assignment
#2
Sara Herrera, Jorge Lora, José I Hormaza, Maria Herrero, Javier Rodrigo
Apricot ( Prunus armeniaca L.) is a species of the Rosaceae that was originated in Central Asia, from where it entered Europe through Armenia. The release of an increasing number of new cultivars from different breeding programs is resulting in an important renewal of plant material worldwide. Although most traditional apricot cultivars in Europe are self-compatible, the use of self-incompatible cultivars as parental genotypes for breeding purposes is leading to the introduction of a number of new cultivars that behave as self-incompatible...
2018: Frontiers in Plant Science
https://www.readbyqxmd.com/read/29715528/functional-analysis-of-duck-goose-and-ostrich-2-5-oligoadenylate-synthetase
#3
Hassan T Tag-El-Din-Hassan, Masami Morimatsu, Takashi Agui
Up-to-date the flavivirus infection in avian taxa is not clearly defined. Several reports have demonstrated that many viruses belonging to Flaviviridae may cause diseases in poultry species; however, the susceptibility of other avian species is variable and still unclear. In human and mice, the 2'-5'-oligoadenylate synthetase (OAS) proteins are associated with resistance to the flavivirus infection as well as other virus infections. However, the avian OAS proteins are rarely studied. In our previous studies, we confirmed that the chicken OAS-like protein (chOASL) expressed OAS-enzymatic activity (the classical OAS/RNase L-dependent pathway) as well as the anti-flavivirus activity (the putative OAS/RNase L-independent pathway)...
April 28, 2018: Infection, Genetics and Evolution
https://www.readbyqxmd.com/read/29671592/inactivation-mechanisms-of-human-and-animal-rotaviruses-by-solar-uva-and-visible-light
#4
Elbashir Araud, Joanna L Shisler, Thanh H Nguyen
Two rotavirus (RV) strains (sialidase-resistant Wa and sialidase-sensitive OSU) were irradiated with simulated solar UVA and visible light in sensitizer-free phosphate buffered solution (lacking exogenous reactive oxygen species (ROS)) or secondary effluent wastewater (producing ROS). Although light attenuated for up to 15% through the secondary effluent wastewater (SEW), the inactivation efficacies increased by 0.7 log10 for Wa and 2 log10 for OSU compared to those in sensitizer-free phosphate buffered solution (PBS) after 4 hrs of irradiation...
April 19, 2018: Environmental Science & Technology
https://www.readbyqxmd.com/read/29659501/the-stability-of-medicinal-plant-micrornas-in-the-herb-preparation-process
#5
Wenyan Xie, Matthias F Melzig
Herbal medicine is now globally accepted as a valid alternative system of pharmaceutical therapies. Various studies around the world have been initiated to develop scientific evidence-based herbal therapies. Recently, the therapeutic potential of medicinal plant derived miRNAs has attracted great attraction. MicroRNAs have been indicated as new bioactive ingredients in medicinal plants. However, the stability of miRNAs during the herbal preparation process and their bioavailability in humans remain unclear...
April 16, 2018: Molecules: a Journal of Synthetic Chemistry and Natural Product Chemistry
https://www.readbyqxmd.com/read/29652922/a-novel-mechanism-of-rnase-l-inhibition-theiler-s-virus-l-protein-prevents-2-5a-from-binding-to-rnase-l
#6
Melissa Drappier, Babal Kant Jha, Sasha Stone, Ruth Elliott, Rong Zhang, Didier Vertommen, Susan R Weiss, Robert H Silverman, Thomas Michiels
The OAS/RNase L pathway is one of the best-characterized effector pathways of the IFN antiviral response. It inhibits the replication of many viruses and ultimately promotes apoptosis of infected cells, contributing to the control of virus spread. However, viruses have evolved a range of escape strategies that act against different steps in the pathway. Here we unraveled a novel escape strategy involving Theiler's murine encephalomyelitis virus (TMEV) L* protein. Previously we found that L* was the first viral protein binding directly RNase L...
April 2018: PLoS Pathogens
https://www.readbyqxmd.com/read/29643235/developing-and-evaluating-inhibitors-against-the-rnase-h-active-site-of-hiv-1-rt
#7
Paul L Boyer, Steven J Smith, Xue Zhi Zhao, Kalyan Das, Kevin Gruber, Eddy Arnold, Terrence R Burke, Stephen H Hughes
We tested three compounds for their ability to inhibit the RNase H (RH) and polymerase activities of HIV-1 reverse transcriptase (RT). A high-resolution crystal structure (2.2 Å) of one of the compounds showed that it chelates the two magnesium ions at the RH active site; this prevents the RH active site from interacting with, and cleaving, the RNA strand of an RNA/DNA heteroduplex. The compounds were tested using a variety of substrates: All three compounds inhibited the polymerase-independent RH activity of HIV-1 RT...
April 11, 2018: Journal of Virology
https://www.readbyqxmd.com/read/29627294/%C3%AE-arrestin-biased-agonism-of-%C3%AE-adrenergic-receptor-regulates-dicer-mediated-microrna-maturation-to-promote-cardioprotective-signaling
#8
Jian-Peng Teoh, Ahmed S Bayoumi, Tatsuya Aonuma, Yanyan Xu, John A Johnson, Huabo Su, Neal L Weintraub, Yaoliang Tang, Il-Man Kim
RATIONALE: MicroRNAs (miRs) are small, non-coding RNAs that function to post-transcriptionally regulate target genes. First transcribed as primary miR transcripts (pri-miRs), they are enzymatically processed by Drosha into premature miRs (pre-miRs) and further cleaved by Dicer into mature miRs. Initially discovered to desensitize β-adrenergic receptor (βAR) signaling, β-arrestins are now well-appreciated to modulate multiple pathways independent of G protein signaling, a concept known as biased signaling...
April 6, 2018: Journal of Molecular and Cellular Cardiology
https://www.readbyqxmd.com/read/29610576/comparison-of-three-pcr-based-assays-for-snp-genotyping-in-plants
#9
Chiara Broccanello, Claudia Chiodi, Andrew Funk, J Mitchell McGrath, Lee Panella, Piergiorgio Stevanato
Background: PCR allelic discrimination technologies have broad applications in the detection of single nucleotide polymorphisms (SNPs) in genetics and genomics. The use of fluorescence-tagged probes is the leading method for targeted SNP detection, but assay costs and error rates could be improved to increase genotyping efficiency. A new assay, rhAmp, based on RNase H2-dependent PCR (rhPCR) combined with a universal reporter system attempts to reduce error rates from primer/primer and primer/probe dimers while lowering costs compared to existing technologies...
2018: Plant Methods
https://www.readbyqxmd.com/read/29606349/angiogenin-ribonuclease-5-is-an-egfr-ligand-and-a-serum-biomarker-for-erlotinib-sensitivity-in-pancreatic-cancer
#10
Ying-Nai Wang, Heng-Huan Lee, Chao-Kai Chou, Wen-Hao Yang, Yongkun Wei, Chun-Te Chen, Jun Yao, Jennifer L Hsu, Cihui Zhu, Haoqiang Ying, Yuanqing Ye, Wei-Jan Wang, Seung-Oe Lim, Weiya Xia, How-Wen Ko, Xiuping Liu, Chang-Gong Liu, Xifeng Wu, Huamin Wang, Donghui Li, Laura R Prakash, Matthew H Katz, Yaan Kang, Michael Kim, Jason B Fleming, David Fogelman, Milind Javle, Anirban Maitra, Mien-Chie Hung
Pancreatic ribonuclease (RNase) is a secreted enzyme critical for host defense. We discover an intrinsic RNase function, serving as a ligand for epidermal growth factor receptor (EGFR), a member of receptor tyrosine kinase (RTK), in pancreatic ductal adenocarcinoma (PDAC). The closely related bovine RNase A and human RNase 5 (angiogenin [ANG]) can trigger oncogenic transformation independently of their catalytic activities via direct association with EGFR. Notably, high plasma ANG level in PDAC patients is positively associated with response to EGFR inhibitor erlotinib treatment...
March 29, 2018: Cancer Cell
https://www.readbyqxmd.com/read/29602068/ectromelia-virus-lacking-the-e3l-ortholog-is-replication-defective-and-nonpathogenic-but-does-induce-protective-immunity-in-a-mouse-strain-susceptible-to-lethal-mousepox
#11
Tiffany R Frey, Katherine S Forsyth, Maura M Sheehan, Brian C De Haven, Julia G Pevarnik, Erin S Hand, Marie C Pizzorno, Laurence C Eisenlohr, Adam R Hersperger
All known orthopoxviruses, including ectromelia virus (ECTV), contain a gene in the E3L family. The protein product of this gene, E3, is a double-stranded RNA-binding protein. It can impact host range and is used by orthopoxviruses to combat cellular defense pathways, such as PKR and RNase L. In this work, we constructed an ECTV mutant with a targeted disruption of the E3L open reading frame (ECTVΔE3L). Infection with this virus resulted in an abortive replication cycle in all cell lines tested. We detected limited transcription of late genes but no significant translation of these mRNAs...
March 27, 2018: Virology
https://www.readbyqxmd.com/read/29587634/high-throughput-detection-of-rna-processing-in-bacteria
#12
Erin E Gill, Luisa S Chan, Geoffrey L Winsor, Neil Dobson, Raymond Lo, Shannan J Ho Sui, Bhavjinder K Dhillon, Patrick K Taylor, Raunak Shrestha, Cory Spencer, Robert E W Hancock, Peter J Unrau, Fiona S L Brinkman
BACKGROUND: Understanding the RNA processing of an organism's transcriptome is an essential but challenging step in understanding its biology. Here we investigate with unprecedented detail the transcriptome of Pseudomonas aeruginosa PAO1, a medically important and innately multi-drug resistant bacterium. We systematically mapped RNA cleavage and dephosphorylation sites that result in 5'-monophosphate terminated RNA (pRNA) using monophosphate RNA-Seq (pRNA-Seq). Transcriptional start sites (TSS) were also mapped using differential RNA-Seq (dRNA-Seq) and both datasets were compared to conventional RNA-Seq performed in a variety of growth conditions...
March 27, 2018: BMC Genomics
https://www.readbyqxmd.com/read/29483516/microrna-contents-in-matrix-vesicles-produced-by-growth-plate-chondrocytes-are-cell-maturation-dependent
#13
Zhao Lin, Michael J McClure, Junjun Zhao, Allison N Ramey, Niels Asmussen, Sharon L Hyzy, Zvi Schwartz, Barbara D Boyan
Chondrocytes at different maturation states in the growth plate produce matrix vesicles (MVs), membrane organelles found in the extracellular matrix, with a wide range of contents, such as matrix processing enzymes and receptors for hormones. We have shown that MVs harvested from growth zone (GC) chondrocyte cultures contain abundant small RNAs, including miRNAs. Here, we determined whether RNA also exists in MVs produced by less mature resting zone (RC) chondrocytes and, if so, whether it differs from the RNA in MVs produced by GC cells...
February 26, 2018: Scientific Reports
https://www.readbyqxmd.com/read/29474813/a-multiplexed-rt-lamp-assay-for-detection-of-group-m-hiv-1-in-plasma-or-whole-blood
#14
Kelly A Curtis, Daphne Morrison, Donna L Rudolph, Anupama Shankar, Laura S P Bloomfield, William M Switzer, S Michele Owen
Isothermal nucleic acid amplification techniques, such as reverse-transcription loop-mediated isothermal amplification (RT-LAMP), exhibit characteristics that are suitable for the development of a rapid, low-cost NAT that can be used at the POC. For demonstration of utility for global use, studies are needed to validate the performance of RT-LAMP for the detection of divergent subtypes. In this study, we designed and evaluated multiplexed HIV-1 integrase RT-LAMP primers to detect subtypes within group M, along with an RNase P positive internal processing and amplification control...
May 2018: Journal of Virological Methods
https://www.readbyqxmd.com/read/29463066/oasl1-traps-viral-rnas-in-stress-granules-to-promote-antiviral-responses
#15
Ji-Seon Kang, Yune-Sahng Hwang, Lark Kyun Kim, Sujung Lee, Wook-Bin Lee, Jeongsil Kim-Ha, Young-Joon Kim
Oligoadenylate synthetase (OAS) protein family is the major interferon (IFN)-stimulated genes responsible for the activation of RNase L pathway upon viral infection. OAS-like (OASL) is also required for inhibition of viral growth in human cells, but the loss of one of its mouse homolog, OASL1, causes a severe defect in termination of type I interferon production. To further investigate the antiviral activity of OASL1, we examined its subcellular localization and regulatory roles in IFN production in the early and late stages of viral infection...
March 31, 2018: Molecules and Cells
https://www.readbyqxmd.com/read/29449323/structural-basis-of-sirna-recognition-by-trbp-double-stranded-rna-binding-domains
#16
Gregoire Masliah, Christophe Maris, Sebastian Lb König, Maxim Yulikov, Florian Aeschimann, Anna L Malinowska, Julie Mabille, Jan Weiler, Andrea Holla, Juerg Hunziker, Nicole Meisner-Kober, Benjamin Schuler, Gunnar Jeschke, Frederic H-T Allain
The accurate cleavage of pre-micro(mi)RNAs by Dicer and mi/siRNA guide strand selection are important steps in forming the RNA-induced silencing complex (RISC). The role of Dicer binding partner TRBP in these processes remains poorly understood. Here, we solved the solution structure of the two N-terminal dsRNA binding domains (dsRBDs) of TRBP in complex with a functionally asymmetric siRNA using NMR, EPR, and single-molecule spectroscopy. We find that siRNA recognition by the dsRBDs is not sequence-specific but rather depends on the RNA shape...
March 15, 2018: EMBO Journal
https://www.readbyqxmd.com/read/29445371/effects-of-microrna-on-regulatory-t-cells-and-implications-for-adoptive-cellular-therapy-to-ameliorate-graft-versus-host-disease
#17
REVIEW
Keli L Hippen, Michael Loschi, Jemma Nicholls, Kelli P A MacDonald, Bruce R Blazar
Regulatory T cells (Tregs) are key mediators of the immune system. MicroRNAs (miRNAs) are a family of ~22 nucleotide non-coding RNAs that are processed from longer precursors by the RNases Drosha and Dicer. miRNA regulates protein expression posttranscriptionally through mRNA destabilization or translational silencing. A critical role for miRNA in Treg function was initially discovered when both Dicer and Drosha knockout (KO) mice were found to develop a fatal autoimmune disease phenotypically similar to Foxp3 KO mice...
2018: Frontiers in Immunology
https://www.readbyqxmd.com/read/29390093/acute-hepatotoxicity-of-2-fluoro-modified-5-10-5-gapmer-phosphorothioate-oligonucleotides-in-mice-correlates-with-intracellular-protein-binding-and-the-loss-of-dbhs-proteins
#18
Wen Shen, Cheryl L De Hoyos, Hong Sun, Timothy A Vickers, Xue-Hai Liang, Stanley T Crooke
We reported previously that a 2' fluoro-modified (2' F) phosphorothioate (PS) antisense oligonucleotides (ASOs) with 5-10-5 gapmer configuration interacted with proteins from Drosophila behavior/human splicing (DBHS) family with higher affinity than PS-ASOs modified with 2'-O-(2-methoxyethyl) (2' MOE) or 2',4'-constrained 2'-O-ethyl (cEt) did. Rapid degradation of these proteins and cytotoxicity were observed in cells treated with 2' F PS-ASO. Here, we report that 2' F gapmer PS-ASOs of different sequences caused reduction in levels of DBHS proteins and hepatotoxicity in mice...
March 16, 2018: Nucleic Acids Research
https://www.readbyqxmd.com/read/29378054/rnase-h2-dependent-polymerase-chain-reaction-and-elimination-of-confounders-in-sample-collection-storage-and-analysis-strengthen-evidence-that-micrornas-in-bovine-milk-are-bioavailable-in-humans
#19
Lanfang Wang, Mahrou Sadri, David Giraud, Janos Zempleni
Background: Evidence suggests that dietary microRNAs (miRs) are bioavailable and regulate gene expression across species boundaries. Concerns were raised that the detection of dietary miRs in plasma might have been due to sample contamination or lack of assay specificity. Objectives: The objectives of this study were to assess potential confounders of plasma miR analysis and to detect miRs from bovine milk in human plasma. Methods: Potential confounders of plasma miR analysis (circadian rhythm, sample collection and storage, calibration, and erythrocyte hemolysis) were assessed by quantitative reverse transcriptase polymerase chain reaction (PCR) by using blood from healthy adults (7 men, 6 women; aged 23-57 y)...
January 1, 2018: Journal of Nutrition
https://www.readbyqxmd.com/read/29297728/chronic-fatigue-and-immune-deficiency-syndrome-cfids-cellular-metabolism-and-ionizing-radiation-a-review-of-contemporary-scientific-literature-and-suggested-directions-for-future-research
#20
Andrej Rusin, Colin Seymour, Carmel Mothersill
PURPOSE: To investigate biochemical pathways known to be involved in radiation response and in CFIDS to determine if there might be common underlying mechanisms leading to symptoms experienced by those accidentally or deliberately exposed to radiation and those suffering from CFIDS. If such a link was established to suggest testable hypotheses to investigate the mechanisms with the aim of identifying new therapeutic targets. CONCLUSIONS: Evidence for involvement of the alpha-synuclein, cytochrome c oxidase, αB-crystallin, RNase L, and lactate dehydrogenase/STAT1 pathways is strong and suggests a common underlying mechanism involving mitochondrial dysfunction mediated by ROS and disruption of ATP production...
March 2018: International Journal of Radiation Biology
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