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https://www.readbyqxmd.com/read/28193027/fluorescence-imaging-of-intracellular-telomerase-activity-using-enzyme-free-signal-amplification
#1
Min Hong, Lidan Xu, Qingwang Xue, Lu Li, Bo Tang
A novel enzyme-free signal amplification-based assay for highly sensitive in situ fluorescence imaging and detection of intracellular telomerase activity was developed by using a gold nanoflare probe-triggered mimic-hybridization chain reaction (mimic-HCR) coupled with a graphene oxide (GO) surface-anchored fluorescence signal readout pathway. The nanoflare probe consists of gold nanoparticles (AuNPs) functionalized with a dense shell of nucleic acid sequences by Au-S bond formation. The nucleic acid sequence is composed of three segments: a long thiol-labeled sequence (HS-DNA) and two short sequences (a telomerase primer sequence, "Primer-DNA", and an FAM-terminated reporter sequence, "Flare-DNA"), both of which are complementary to HS-DNA...
December 20, 2016: Analytical Chemistry
https://www.readbyqxmd.com/read/28190655/development-of-helix-stabilized-cell-penetrating-peptides-containing-cationic-%C3%AE-%C3%AE-disubstituted-amino-acids-as-helical-promoters
#2
Hiroko Yamashita, Takashi Misawa, Makoto Oba, Masakazu Tanaka, Mikihiko Naito, Masaaki Kurihara, Yosuke Demizu
Cell-penetrating peptides (CPP) have attracted many scientists' attention as intracellular delivery tools due to their high cargo molecule transportation efficiency and low cytotoxicity. Therefore, in many research fields CPP, such as HIV-Tat and oligoarginine (Rn), are used to deliver hydrophilic drugs and biomolecules, including proteins, DNA, and RNA. We designed four types of CPP that contained cationic α,α-disubstituted amino acids (Api(C2Gu) and Api(C4Gu)) as helical promoters; i.e., 1-4 [FAM-β-Ala-(l-Arg-l-Arg-Xaa)3-(Gly)3-NH2 (1: Xaa=Api(C2Gu), 2: Xaa=Api(C4Gu)), 3: FAM-β-Ala-(l-Arg)8-Api(C2Gu)-(Gly)3-NH2, and 4: FAM-β-Ala-(l-Arg)5-Api(C2Gu)-(l-Arg)2-Api(C2Gu)-(Gly)3-NH2], and investigated their preferred secondary structures and cell membrane-penetrating ability...
February 1, 2017: Bioorganic & Medicinal Chemistry
https://www.readbyqxmd.com/read/28187297/a-universal-aptameric-biosensor-multiplexed-detection-of-small-analytes-via-aggregated-perylene-based-broad-spectrum-quencher
#3
Rong Hu, Xi Zhang, Qiang Xu, Dan-Qing Lu, Yun-Hui Yang, Quan-Qing Xu, Qiong Ruan, Liu-Ting Mo, Xiao-Bing Zhang
A universal aptameric system based on the taking advantage of double-stranded DNA/perylene diimide (dsDNA/PDI) as the signal probe was developed for multiplexed detection of small molecules. Aptamers are single-stranded DNA or RNA oligonucleotides which are selected in vitro by a process known as systematic evolution of ligands by exponential enrichment. In this work, we synthesized a new kind of PDI and reported this aggregated PDI could quench the double-stranded DNA (dsDNA)-labeled fluorophores with a high quenching efficiency...
January 24, 2017: Biosensors & Bioelectronics
https://www.readbyqxmd.com/read/28159568/micro-rna-detection-based-on-fluorescence-resonance-energy-transfer-of-dna-carbon-quantum-dots-probes
#4
Faeze Khakbaz, Mohamad Mahani
Carbon quantum dots have been proposed as an effective platform for miRNA detection. Carbon dots were synthesized by citric acid. The synthesized dots were characterized by dynamic light scattering, UV-Vis spectrophotometry, spectrofluorimetry, transmission electron microscopy and FT-IR spectrophotometry. The fluorescence quantum yield of the synthesized dots was determined using quinine sulfate as the standard. The FAM-labeled single stranded DNA, as sensing element, was adsorbed on dots by π-π interaction...
January 31, 2017: Analytical Biochemistry
https://www.readbyqxmd.com/read/28142223/cryphonectria-parasitica-the-causal-agent-of-chestnut-blight-invasion-history-population-biology-and-disease-control
#5
Daniel Rigling, Simone Prospero
Chestnut blight, caused by Cryphonectria parasitica, is a devastating disease infecting American and European chestnut trees. The pathogen is native to East Asia and was spread to other continents via infected chestnut plants. This review summarizes the current state of research on this pathogen with a special emphasis on its interaction with a hyperparasitic mycovirus that acts as a biological control agent of chestnut blight. Taxonomy: Cryphonectria parasitica (Murr.) Barr. is a Sordariomycete (ascomycete) fungus in the family Cryphonetriaceae (Order Diaporthales)...
January 31, 2017: Molecular Plant Pathology
https://www.readbyqxmd.com/read/28140505/n-2-substituted-2-deoxyguanosine-triphosphate-derivatives-as-selective-substrates-for-human-dna-polymerase-%C3%AE%C2%BA
#6
A S Prakasha Gowda, Marietta Lee, Thomas E Spratt
N(2) -Alkyl-2'-deoxyguanosine triphosphate (N(2) -alkyl-dGTP) derivatives with methyl, butyl, benzyl, or 4-ethynylbenzyl substituents were prepared and tested as substrates for human DNA polymerases. N(2) -Benzyl-dGTP was equal to dGTP as a substrate for DNA polymerase κ (pol κ), but was a poor substrate for pols β, δ, η, ι, or ν. In vivo reactivity was evaluated through incubation of N(2) -4-ethynylbenzyl-dG with wild-type and pol κ deficient mouse embryonic fibroblasts. CuAAC reaction with 5(6)-FAM-azide demonstrated that only cells containing pol κ were able to incorporate N(2) -4-ethynylbenzyl-dG into the nucleus...
January 31, 2017: Angewandte Chemie
https://www.readbyqxmd.com/read/28139463/microfluidic-chip-based-micro-rna-detection-through-the-combination-of-fluorescence-and-surface-enhanced-raman-scattering-techniques
#7
Zhile Wang, Shenfei Zong, Zhuyuan Wang, Lei Wu, Peng Chen, Binfeng Yun, Yiping Cui
We present a novel microfluidic chip based method for the detection of micro RNA (miRNA) via the combination of fluorescence and surface enhanced Raman scattering (SERS) spectroscopies. First, silver nanoparticles (Ag NPs) are immobilized onto a glass slide, forming a SERS enhancing substrate. Then a specificially designed molecular beacon (MB) is attached to the SERS substrate. The 3' end of the MB is decorated with a thiol group to facilitate the attachment of the MB, while the 5' end of the MB is labeled with an organic dye 6-FAM, which is used both as the fluorophore and SERS reporter...
March 10, 2017: Nanotechnology
https://www.readbyqxmd.com/read/28138336/chemical-compositions-chromatographic-fingerprints-and-antioxidant-activities-of-citri-exocarpium-rubrum-juhong
#8
Yang Zhao, Chun-Pin Kao, Chi-Ren Liao, Kun-Chang Wu, Xin Zhou, Yu-Ling Ho, Yuan-Shiun Chang
BACKGROUND: Citri Exocarpium Rubrum (CER), which is known as Juhong in Chinese, is the dried exocarp of Citrus reticulata Blanco and its cultivars (Fam. Rutaceae) and is currently used in Chinese medicine to protect the stomach and eliminate dampness and phlegm. The main aim of this study was to develop a high-performance liquid chromatography ultraviolet mass spectrometry (HPLC-UV-MS) method for determining the chemical compositions and fingerprint of CER. We also evaluated the antioxidant properties of CER based on its 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging activity, ferric ion reducing antioxidant power (FRAP) and trolox equivalent antioxidant capacity (TEAC) assays...
2017: Chinese Medicine
https://www.readbyqxmd.com/read/28131696/investigating-fam-n-pulses-for-signal-enhancement-in-mqmas-nmr-of-quadrupolar-nuclei
#9
Henri Colaux, Daniel M Dawson, Sharon E Ashbrook
Although a popular choice for obtaining high-resolution solid-state NMR spectra of quadrupolar nuclei, the inherently low sensitivity of the multiple-quantum magic-angle spinning (MQMAS) experiment has limited its application for nuclei with low receptivity or when the available sample volume is limited. A number of methods have been introduced in the literature to attempt to address this problem. Recently, we have introduced an alternative, automated approach, based on numerical simulations, for generating amplitude-modulated pulses (termed FAM-N pulses) to enhance the efficiency of the triple- to single-quantum conversion step within MQMAS...
January 18, 2017: Solid State Nuclear Magnetic Resonance
https://www.readbyqxmd.com/read/28131500/resurfacing-the-penis-of-complex-hypospadias-repair-hypospadias-cripples
#10
Mina M Fam, Moneer K Hanna
PURPOSE: After the creation of a neourethra in a "hypospadias cripple," resurfacing the penis with healthy skin is a significant challenge because local tissue is often scarred and unusable. We reviewed our experience with various strategies to resurface the penis of hypospadias cripples. MATERIALS AND METHODS: We retrospectively reviewed the records of 215 patients referred after multiple unsuccessful hypospadias repairs from 1981 to 2014. In 130 of 215 patients we performed resurfacing using local penile flaps using various techniques, including Byars flaps, Z-plasty or double Z-plasty, or a dorsal relaxing incision...
January 25, 2017: Journal of Urology
https://www.readbyqxmd.com/read/28124559/biodegradable-mno2-nanosheet-mediated-signal-amplification-in-living-cells-enables-sensitive-detection-of-down-regulated-intracellular-microrna
#11
Jing Li, Daxiu Li, Ruo Yuan, Yun Xiang
The monitoring of intracellular microRNAs plays important roles in elucidating the biological function and biogenesis of miRNAs in living cells. However, because of their sequence similarity, low abundance and small size, it is of great challenge to detect intracellular miRNAs, especially for those with much lower expression levels. To address this issue, we have developed an in cell signal amplification approach for monitoring down-regulated miRNAs in living cells based on biodegradable MnO2 nanosheet-mediated and target-triggered assembly of hairpins...
January 26, 2017: ACS Applied Materials & Interfaces
https://www.readbyqxmd.com/read/28114894/evolutional-dynamics-of-45s-and-5s-ribosomal-dna-in-ancient-allohexaploid-atropa-belladonna
#12
Roman A Volkov, Irina I Panchuk, Nikolai V Borisjuk, Marta Hosiawa-Baranska, Jolanta Maluszynska, Vera Hemleben
BACKGROUND: Polyploid hybrids represent a rich natural resource to study molecular evolution of plant genes and genomes. Here, we applied a combination of karyological and molecular methods to investigate chromosomal structure, molecular organization and evolution of ribosomal DNA (rDNA) in nightshade, Atropa belladonna (fam. Solanaceae), one of the oldest known allohexaploids among flowering plants. Because of their abundance and specific molecular organization (evolutionarily conserved coding regions linked to variable intergenic spacers, IGS), 45S and 5S rDNA are widely used in plant taxonomic and evolutionary studies...
January 23, 2017: BMC Plant Biology
https://www.readbyqxmd.com/read/28107953/visual-detection-of-nucleic-acids-based-on-lateral-flow-biosensor-and-hybridization-chain-reaction-amplification
#13
Na Ying, Chuanjing Ju, Zhongyi Li, Wensen Liu, Jiayu Wan
In this study, a new lateral flow nucleic acid biosensor (LFNAB) using hybridization chain reaction (HCR) for signal amplification was developed for visual detection of nucleic acids with high sensitivity and low cost. A "sandwich-type" detection strategy was employed in our design. The sandwich system of capture probe (CP)/target DNA/reporter probe (RP)-HCR complexes was fabricated as the sensing platform. As the initiator strand, reporter probe propagated a chain reaction of hybridization events between the two hairpin probes modified with biotin, and determined whether long nicked DNA polymers were formed...
March 1, 2017: Talanta
https://www.readbyqxmd.com/read/28099303/quality-of-life-in-bilateral-vs-unilateral-end-stage-ankle-arthritis-and-outcomes-of-bilateral-vs-unilateral-total-ankle-replacement
#14
Sagar J Desai, Mark Glazebrook, Murray J Penner, Kevin J Wing, Alastair S E Younger, Ellie Pinsker, Timothy R Daniels
BACKGROUND: We performed a retrospective cohort study to compare preoperative health-related quality of life (HRQoL) between patients with bilateral and those with unilateral end-stage ankle arthritis. We also compared midterm outcomes in a subgroup of patients who had undergone staged bilateral total ankle replacement (TAR) with the outcomes in the group treated with unilateral TAR. METHODS: The HRQoL before surgical treatment was compared between 53 patients with bilateral end-stage ankle arthritis identified from the Canadian Orthopaedic Foot and Ankle Society Prospective Ankle Reconstruction Database and 106 patients with unilateral arthritis selected from the same database...
January 18, 2017: Journal of Bone and Joint Surgery. American Volume
https://www.readbyqxmd.com/read/28091705/bacteria-of-the-family-candidatus-midichloriaceae-in-sympatric-zones-of-ixodes-ticks-genetic-evidence-for-vertical-transmission
#15
Tatyana A Mukhacheva, Sergey Y Kovalev
Ixodes ticks transmit infectious agents and also harbor their own parasites and symbionts. The presumptive endosymbiont of Ixodes ricinus, 'Candidatus Midichloria mitochondrii', has a unique ability to invade mitochondria within tick ovarian cells and is transovarially transmitted with 100% efficiency. A closely related bacterium, provisionally named Montezuma (now 'Candidatus Lariskella arthropodarum'), was isolated from the Ixodes persulcatus ticks and human blood in 2004 as well as from Ixodes pavlovskyi in 2015...
January 14, 2017: Microbial Ecology
https://www.readbyqxmd.com/read/28079361/controllable-synthesis-of-a-smart-multifunctional-nanoscale-metal-organic-framework-for-magnetic-resonance-optical-imaging-and-targeted-drug-delivery
#16
Xuechuan Gao, Manjue Zhai, Weihua Guan, Jingjuan Liu, Zhiliang Liu, Alatangaole Damirin
As a result of their extraordinarily large surfaces and well-defined pores, the design of a multifunctional metal-organic framework (MOF) is crucial for drug delivery but has rarely been reported. In this paper, a novel drug delivery system (DDS) based on nanoscale MOF was developed for use in cancer diagnosis and therapy. This MOF-based tumor targeting DDS was fabricated by a simple postsynthetic surface modification process. First, magnetic mesoporous nanomaterial Fe-MIL-53-NH2 was used for encapsulating the drug and served as a magnetic resonance contrast agent...
February 1, 2017: ACS Applied Materials & Interfaces
https://www.readbyqxmd.com/read/28062406/anatomically-preserved-strobili-and-leaves-from-the-permian-of-china-dorsalistachyaceae-fam-nov-broaden-knowledge-of-noeggerathiales-and-constrain-their-possible-taxonomic-affinities
#17
Shi-Jun Wang, Richard M Bateman, Alan R T Spencer, Jun Wang, Longyi Shao, Jason Hilton
PREMISE OF THE STUDY: Noeggerathiales are an extinct group of heterosporous shrubs and trees that were widespread and diverse during the Pennsylvanian-Permian Epochs (323-252 Ma) but are of controversial taxonomic affinity. Groups proposed as close relatives include leptosporangiate ferns, sphenopsids, progymnosperms, or the extant eusporangiate fern Tmesipteris. Previously identified noeggerathialeans lacked anatomical preservation, limiting taxonomic comparisons to their external morphology and spore structure...
January 6, 2017: American Journal of Botany
https://www.readbyqxmd.com/read/28056380/shedding-light-on-the-polyphyletic-thecate-amoeba-genus-plagiophrys-transition-of-some-of-its-species-to-rhizaspis-tectofilosida-thecofilosea-cercozoa-and-the-establishment-of-sacciforma-gen-nov-and-rhogostomidae-fam-nov-cryomonadida-thecofilosea-cercozoa
#18
Kenneth Dumack, Hüsna Öztoprak, Lioba Rüger, Michael Bonkowski
For over a century testate amoebae have been a favoured group of interest for protistologists, however there is still an endless amount of unanswered questions. The genus Plagiophrys, Claparède and Lachmann 1859, is still one of the unresolved mysteries as it comprises species with high morphological diversity of which no molecular data are available. To shed light on the phylogeny and taxonomy of Plagiophrys we (a) cultured four isolates of three Plagiophrys morphospecies and provided morphological observations (b) obtained three new SSU RNA gene sequences and conducted phylogenetic analyses of the Thecofilosea and (c) did intensive literature research, showing that Plagiophrys is polyphyletic...
December 7, 2016: Protist
https://www.readbyqxmd.com/read/28055181/fluorescent-branched-rnas-for-high-throughput-analysis-of-dbr1-enzyme-kinetics-and-inhibition
#19
Adam Katolik, Nathaniel E Clark, Nobuhiro Tago, Eric J Montemayor, P John Hart, Masad J Damha
We have developed fluorescent 2',5' branched RNAs (bRNA) that permit real time monitoring of RNA lariat (intron) debranching enzyme (Dbr1) kinetics. These compounds contain fluorescein (FAM) on the 5' arm of the bRNA that is quenched by a dabcyl moiety on the 2' arm. Dbr1-mediated hydrolysis of the 2',5' linkage induces a large increase in fluorescence, providing a convenient assay for Dbr1 hydrolysis. We show that unlabeled bRNAs with non-native 2',5'-phosphodiester linkages, such as phosphoramidate or phosphorothioate, can inhibit Dbr1-mediated debranching with IC50 values in the low nanomolar range...
January 18, 2017: ACS Chemical Biology
https://www.readbyqxmd.com/read/28054727/direct-comparison-of-point-by-point-and-rapid-ultra-high-resolution-electroanatomical-mapping-in-patients-scheduled-for-ablation-of-atrial-fibrillation
#20
Laura Rottner, Andreas Metzner, Feifan Ouyang, Christian Heeger, Kentaro Hayashi, Thomas Fink, Christine Lemes, Shibu Mathew, Tilman Maurer, Bruno REIßMANN, Enida Rexha, Johannes Riedl, Ardan M Saguner, Francesco Santoro, Karl-Heinz Kuck, Christian Sohns
INTRODUCTION: Three-dimensional electroanatomical mapping (EAM) is an established tool facilitating catheter ablation. In this context, the novel Rhythmia system sets a new bar in fast high-resolution mapping. The aim of this study was to directly compare point-by-point versus rapid ultra-high-resolution EAM in patients scheduled for ablation of atrial fibrillation (AF) with focus on procedural data, acute success, and midterm clinical outcome. METHODS AND RESULTS: A total number of 74 consecutive patients (48/74 male) with symptomatic AF were scheduled to undergo pulmonary vein isolation (PVI) using either Carto or Rhythmia...
January 5, 2017: Journal of Cardiovascular Electrophysiology
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