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https://www.readbyqxmd.com/read/29792924/creation-of-an-acellular-vaginal-matrix-for-potential-vaginal-augmentation-and-cloacal-repair
#1
K V Greco, L G Jones, I Obiri-Yeboa, T Ansari
STUDY OBJECTIVE: our aim was to use porcine vagina to create a vaginal matrix and test its cellular biocompatibility. DESIGN: vagina was harvested from pigs and de-cellularised (DC) using a combination of detergents (Triton x-100 and sodium deoxycholate) and enzymes (DNAse/RNAse). INTERVENTIONS: the presence of cellular material, collagen structural integrity and basement membrane proteins were assessed histologically. To address cytocompatibility, porcine adipose derived-mesenchymal stem cells (AD-MSC) were harvested from abdominal fat together with vaginal epithelial cells (VEC) and seeded onto the mucosal aspect of the vaginal scaffold...
May 21, 2018: Journal of Pediatric and Adolescent Gynecology
https://www.readbyqxmd.com/read/29792692/small-molecule-targeted-recruitment-of-a-nuclease-to-rna
#2
Matthew G Costales, Yasumasa Matsumoto, Sai Pradeep Velagapudi, Matthew D Disney
The choreography between RNA synthesis and degradation is a key determinant in biology. Engineered systems such as CRISPR have been developed to rid a cell of RNAs. Here, we show that a small molecule can recruit a nuclease to a specific transcript, triggering its destruction. A small molecule that selectively binds the oncogenic microRNA(miR)-96 hairpin precursor was appended with a short 2'-5' poly(A) oligonucleotide. The conjugate locally activated endogenous, latent ribonuclease (RNase L), which selectively cleaved the miR-96 precursor in cancer cells in a catalytic and sub-stoichiometric fashion...
May 24, 2018: Journal of the American Chemical Society
https://www.readbyqxmd.com/read/29790953/identifying-and-avoiding-off-target-effects-of-rnase-h-dependent-antisense-oligonucleotides-in-mice
#3
Peter H Hagedorn, Malene Pontoppidan, Tina S Bisgaard, Marco Berrera, Andreas Dieckmann, Martin Ebeling, Marianne R Møller, Heidi Hudlebusch, Marianne L Jensen, Henrik F Hansen, Troels Koch, Morten Lindow
Antisense oligonucleotides that are dependent on RNase H for cleavage and subsequent degradation of complementary RNA are being developed as therapeutics. Besides the intended RNA target, such oligonucleotides may also cause degradation of unintended RNA off-targets by binding to partially complementary target sites. Here, we characterized the global effects on the mouse liver transcriptome of four oligonucleotides designed as gapmers, two targeting Apob and two targeting Pcsk9, all in different regions on their respective intended targets...
May 22, 2018: Nucleic Acids Research
https://www.readbyqxmd.com/read/29790504/molecularly-imprinted-nanoparticles-for-inhibiting-ribonuclease-in-reverse-transcriptase-polymerase-chain-reaction
#4
Xiaotong Feng, Jon Ashley, Tongchang Zhou, Arnab Halder, Yi Sun
Molecularly imprinted nanoparticles (nanoMIPs) are synthesized via a solid-phase approach using RNase as the template. The feasibility of employing the nanoMIPs as RNase inhibitor is successfully demonstrated in reverse transcriptase polymerase chain reaction (RT-PCR) assays, suggesting the tailor-made nanomaterials are very promising for use in routine biological assays.
May 23, 2018: Analyst
https://www.readbyqxmd.com/read/29789788/development-of-two-analytical-methods-based-on-reverse-phase-chromatographic-and-sds-page-gel-for-assessment-of-deglycosylation-yield-in-n-glycan-mapping
#5
Anahita D Eckard, David R Dupont, Johnie K Young
N -lined glycosylation is one of the critical quality attributes (CQA) for biotherapeutics impacting the safety and activity of drug product. Changes in pattern and level of glycosylation can significantly alter the intrinsic properties of the product and, therefore, have to be monitored throughout its lifecycle. Therefore fast, precise, and unbiased N -glycan mapping assay is desired. To ensure these qualities, using analytical methods that evaluate completeness of deglycosylation is necessary. For quantification of deglycosylation yield, methods such as reduced liquid chromatography-mass spectrometry (LC-MS) and reduced capillary gel electrophoresis (CGE) have been commonly used...
2018: BioMed Research International
https://www.readbyqxmd.com/read/29779946/pnpt1-release-from-mitochondria-during-apoptosis-triggers-decay-of-poly-a-rnas
#6
Xing Liu, Rui Fu, Youdong Pan, Karla F Meza-Sosa, Zhibin Zhang, Judy Lieberman
Widespread mRNA decay, an unappreciated feature of apoptosis, enhances cell death and depends on mitochondrial outer membrane permeabilization (MOMP), TUTases, and DIS3L2. Which RNAs are decayed and the decay-initiating event are unknown. Here, we show extensive decay of mRNAs and poly(A) noncoding (nc)RNAs at the 3' end, triggered by the mitochondrial intermembrane space 3'-to-5' exoribonuclease PNPT1, released during MOMP. PNPT1 knockdown inhibits apoptotic RNA decay and reduces apoptosis, while ectopic expression of PNPT1, but not an RNase-deficient mutant, increases RNA decay and cell death...
May 10, 2018: Cell
https://www.readbyqxmd.com/read/29779770/normalised-quantitative-polymerase-chain-reaction-for-diagnosis-of-tuberculosis-associated-uveitis
#7
Manas Ranjan Barik, Soveeta Rath, Rohit Modi, Rajkishori Rana, Mamatha M Reddy, Soumyava Basu
Polymerase chain reaction (PCR)-based diagnosis of tuberculosis-associated uveitis (TBU) in TB-endemic countries is challenging due to likelihood of latent mycobacterial infection in both immune and non-immune cells. In this study, we investigated normalised quantitative PCR (nqPCR) in ocular fluids (aqueous/vitreous) for diagnosis of TBU in a TB-endemic population. Mycobacterial copy numbers (mpb64 gene) were normalised to host genome copy numbers (RNAse P RNA component H1 [RPPH1] gene) in TBU (n = 16) and control (n = 13) samples (discovery cohort)...
May 2018: Tuberculosis
https://www.readbyqxmd.com/read/29777050/the-yeast-telomerase-recruitment-module-requires-a-specific-rna-architecture
#8
Nancy Laterreur, Bruno Lemieux, Hannah Neumann, Jean-Christophe Berger-Dancause, Daniel Lafontaine, Raymund J Wellinger
Telomerases are ribonucleoprotein (RNP) enzymes that are related to reverse transcriptases. While they maintain genome stability, their composition varies significantly between species. Yeast telomerase RNPs contain an RNA that is comparatively large and its overall folding shows long helical segments with distal functional parts. Here we investigated the essential stem IVc module of the budding yeast telomerase RNA, called Tlc1. The distal part of stem IVc includes a conserved sequence element CS2a and structurally conserved features to which bind the Pop1/Pop6/Pop7 proteins and which together function analogously to the P3 domains of the RNase P/MRP RNPs...
May 18, 2018: RNA
https://www.readbyqxmd.com/read/29776310/preparation-and-evaluation-of-ribonuclease-resistant-viral-hiv-rna-standards-based-on-armored-rna-technology
#9
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/29775224/circular-rnas-are-differentially-expressed-in-liver-ischemia-reperfusion-injury-model
#10
Zhiqiang Ye, Qinglei Kong, Jianhua Han, Jingyi Deng, Miaolue Wu, Hong Deng
Liver ischemia/reperfusion (I/R) injury has high mortality due to the intense inflammatory process occurs in the liver. However, the pathological mechanism underlying I/R injury is still not clear. Recent works showed that circular RNAs play critical roles in many human diseases. In this study, the occurrence of liver I/R injury was validated by an analysis of the blood samples and hematoxylin-eosin (HE) staining of liver tissues. Total RNA was purified and followed by RNA-seq in the purpose of screening the circRNAs in significant differentially expression, which were validated by quantitative PCR...
May 18, 2018: Journal of Cellular Biochemistry
https://www.readbyqxmd.com/read/29773824/rnase-h-assisted-rna-primed-rolling-circle-amplification-for-targeted-rna-sequence-detection
#11
Hirokazu Takahashi, Masahiko Ohkawachi, Kyohei Horio, Toshiro Kobori, Tsunehiro Aki, Yukihiko Matsumura, Yutaka Nakashimada, Yoshiko Okamura
RNA-primed rolling circle amplification (RPRCA) is a useful laboratory method for RNA detection; however, the detection of RNA is limited by the lack of information on 3'-terminal sequences. We uncovered that conventional RPRCA using pre-circularized probes could potentially detect the internal sequence of target RNA molecules in combination with RNase H. However, the specificity for mRNA detection was low, presumably due to non-specific hybridization of non-target RNA with the circular probe. To overcome this technical problem, we developed a method for detecting a sequence of interest in target RNA molecules via RNase H-assisted RPRCA using padlocked probes...
May 17, 2018: Scientific Reports
https://www.readbyqxmd.com/read/29772476/fate-of-nuclear-material-during-subsequent-steps-of-the-kinetin-induced-pcd-in-apical-parts-of-vicia-faba-ssp-minor-seedling-roots
#12
Andrzej Kaźmierczak, Kamila Soboska
In animals during apoptosis, the best examined type of programmed cell death (PCD), three main phases are distinguished: (i) specification (signaling), (ii) killing and (iii) execution one. It has bean postulated that plant PCD also involves three subsequent phases: (i) transmission of death signals to cells (signaling), (ii) initiation of killing processes and (iii) destruction of cells. One of the most important hallmarks of animal and plant PCD are those regarding nucleus, not thoroughly studied in plants so far...
April 30, 2018: Micron: the International Research and Review Journal for Microscopy
https://www.readbyqxmd.com/read/29769336/cytoplasmic-re-localization-and-colocalization-with-viroplasms-of-host-cell-proteins-and-their-role-in-rotavirus-infection
#13
Poonam Dhillon, Varsha N Tandra, Sandip G Chorghade, Nima D Namsa, Lipika Sahoo, C Durga Rao
Rotavirus replicates in the cytoplasm of infected cells in unique virus-induced cytoplasmic inclusion bodies called viroplasms (VM), which are nucleated by two essential viral non-structural proteins NSP2 and NSP5. However, the precise composition of the VM, intracellular localization of host proteins during virus infection, their association with VM or role in rotavirus growth remained largely unexplored. Mass-spectrometry analyses revealed the presence of several host heterogeneous nuclear ribonucleoproteins (hnRNPs) and AU-rich element-binding proteins (ARE-BPs) and cytoplasmic proteins from uninfected MA104 cell extracts in the pull-down (PD) complexes of purified viroplasmic proteins NSP2 and NSP5...
May 16, 2018: Journal of Virology
https://www.readbyqxmd.com/read/29764511/high-serum-levels-of-pregenomic-rna-reflect-frequently-failing-reverse-transcription-in-hepatitis-b-virus-particles
#14
Kasthuri Prakash, Gustaf E Rydell, Simon B Larsson, Maria Andersson, Gunnar Norkrans, Heléne Norder, Magnus Lindh
BACKGROUND: Hepatocytes infected by hepatitis B virus (HBV) produce different HBV RNA species, including pregenomic RNA (pgRNA), which is reverse transcribed during replication. Particles containing HBV RNA are present in serum of infected individuals, and quantification of this HBV RNA could be clinically useful. METHODS: In a retrospective study of 95 patients with chronic HBV infection, we characterised HBV RNA in serum in terms of concentration, particle association and sequence...
May 15, 2018: Virology Journal
https://www.readbyqxmd.com/read/29762514/blackcurrant-leaf-chlorosis-associated-virus-evidence-of-the-presence-of-circular-rna-during-infections
#15
Delano James, James Phelan, Daniel Sanderson
Blackcurrant leaf chlorosis associated virus (BCLCaV) was detected recently by next-generation sequencing (NGS) and a new and distinct species in the genus Idaeovirus was proposed. Analysis of NGS-derived paired-end reads revealed the existence of bridge reads encompassing the 3'-terminus and 5'-terminus of RNA-2 or RNA-3 of BCLCaV. The full RNA-2 or RNA-3 could be amplified using outward facing or abutting primers; also, RNA-2/RNA-3 could be detected even after three consecutive RNase R enzyme treatments, with denaturation at 95 °C preceding each digestion...
May 15, 2018: Viruses
https://www.readbyqxmd.com/read/29758474/purification-and-characterization-of-two-pathogenesis-related-class-10-protein-isoforms-with-ribonuclease-activity-from-the-fresh-angelica-sinensis-roots
#16
Jianru Pan, Xiangling Wang, Lingling Li, Xian Li, Xiaoqiang Ye, Di Lv, Cuihuang Chen, Shutao Liu, Huocong He
In this study, two pathogenesis-related (PR) class 10 protein isoforms, ASPR-1 and ASPR-2, were purified from fresh roots of the Chinese medicinal plant Angelica sinensis (A. sinensis) using 80% ammonium sulfate precipitation, Sephadex G50 gel filtration chromatography, and DEAE-Sepharose ion-exchange chromatography. The molecular masses of ASPR-1 and ASPR-2 were estimated to be 16.66 kDa and 16.46 kDa, respectively, using sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The isoforms are both glycoproteins containing glycosyl contents of 1...
April 30, 2018: Plant Physiology and Biochemistry: PPB
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
#17
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/29753166/double-determination-of-long-noncoding-rnas-from-lung-cancer-via-multi-amplified-electrochemical-genosensor-at-sub-femtomole-level
#18
Xiaoyan Li, Gang Peng, Feng Cui, Qianying Qiu, Xiaojun Chen, He Huang
An ultrasensitive electrochemical genosensor has been fabricated for the double determination of two different specific sequences deduced from the maternally expressed gene3 (MEG3) lncRNA (long noncoding RNA), which was demonstrated by coupling RNase A-aided target recycling with DNA supersandwich-induced signal enhancement, based on a composite interface of graphene-like tungsten disulfide/dendritic gold nanostructures (WS2 /DGN). Firstly, duple target sequences of T1 and T2 were captured by the primer probes of P1 /P2 functionalized Fe3 O4 @C magnetic nanoparticles, via the DNA/RNA hybridization between T1 /T2 and P1 /P2 ...
May 2, 2018: Biosensors & Bioelectronics
https://www.readbyqxmd.com/read/29751065/a-3-utr-derived-non-coding-rna-ribs-increases-expression-of-cfa-and-promotes-biofilm-formation-of-salmonella-enterica-serovar-typhi
#19
Xin Zhao, Rui Liu, Hao Tang, George Osei-Adjei, Shungao Xu, Ying Zhang, Xinxiang Huang
Bacterial non-coding RNAs (ncRNAs) are widely studied and found to play important roles in regulating various cellular processes. Recently, many ncRNAs have been discovered to be transcribed or processed from 3' untranslated regions (3' UTRs). Here we reported a novel 3' UTR-derived ncRNA, RibS, which could influence biofilm formation of Salmonella enterica serovar Typhi (S. Typhi). RibS was confirmed to be a ∼700 nt processed product produced by RNase III-catalyzed cleavage from the 3' UTR of riboflavin synthase subunit alpha mRNA, RibE...
May 8, 2018: Research in Microbiology
https://www.readbyqxmd.com/read/29738685/blood-derived-rna-and-microrna-hydrolyzing-igg-antibodies-in-schizophrenia-patients
#20
E A Ermakov, S A Ivanova, V N Buneva, G A Nevinsky
Abzymes with various catalytic activities are the earliest statistically significant markers of existing and developing autoimmune diseases (AIDs). Currently, schizophrenia (SCZD) is not considered to be a typical AID. It was demonstrated recently that antibodies from SCZD patients efficiently hydrolyze DNA and myelin basic protein. Here, we showed for the first time that autoantibodies from 35 SCZD patients efficiently hydrolyze RNA (cCMP > poly(C) > poly(A) > yeast RNA) and analyzed site-specific hydrolysis of microRNAs involved in the regulation of several genes in SCZD (miR-137, miR-9-5p, miR-219-2-3p, and miR-219a-5p)...
May 2018: Biochemistry. Biokhimii︠a︡
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