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Tagging and capture

Xian-Feng Jiang, Xing-Fu Zhu, Qing-Jun Li
Distyly is a widespread floral polymorphism characterized by the flowers within a population showing reciprocal placement of the anthers and stigma. Darwin hypothesizes that distyly evolves to promote precise pollen transfer between morphs. Primula chungensis exhibits two types of anther heights, and these two types of anthers show pollen of two different size classes. To understand whether the stigma could capture more pollen grains from the anthers of the pollen donor as the separation between the stigma of pollen receiver and the anther of pollen donor decreased, the present research assessed the source of the pollen load in a series of open-pollinated flowers with continuous variation of style lengths...
June 2018: AoB Plants
Yuqing Zhang, Gongwei Sun, Yi Zhang, Biao Huang, Zhi Xing, Sichun Zhang, Xinrong Zhang
Inductively Coupled Plasma Mass Spectrometry (ICP-MS) based immunoassay method has been proposed in multiple immunoassays but has not been used in competitive and sandwich formats immunoassay simultaneously. The two immunoassays were usually conducted separately in clinical field depending on the size and the amount of binding sites of targets. We proposed an immunoassay method based on magnetic beads and ICP-MS detection that could be suitable for both small and large molecules. The functionalized magnetic beads were added to capture the immune complex after immune reaction...
August 1, 2018: Talanta
Tin Ki Tsang, Eric A Bushong, Daniela Boassa, Junru Hu, Benedetto Romoli, Sebastien Phan, Davide Dulcis, Chih-Ying Su, Mark H Ellisman
Electron microscopy (EM) offers unparalleled power to study cell substructures at the nanoscale. Cryofixation by high-pressure freezing offers optimal morphological preservation, as it captures cellular structures instantaneously in their near-native states. However, the applicability of cryofixation is limited by its incompatibilities with diaminobenzidine labeling using genetic EM tags and the high-contrast en bloc staining required for serial block-face scanning electron microscopy (SBEM). In addition, it is challenging to perform correlated light and electron microscopy (CLEM) with cryofixed samples...
May 11, 2018: ELife
Sandra Sáez-Durán, Ángela L Debenedetti, Sandra Sainz-Elipe, M Teresa Galán-Puchades, Màrius V Fuentes
Serra Calderona Natural Park, a Mediterranean ecosystem, has been in post-fire regeneration for 10 years. To elucidate which helminth community component species of the wood mouse, Apodemus sylvaticus, can be considered biological tags of this process, the influence of intrinsic (host density; host sex and age) and extrinsic factors (site, year, and period of capture; vegetation recovery) on their prevalence and abundance has been analysed, comparing a burned and an unburned area. A total of 564 wood mice (408 from the burned and 156 from the unburned area), from the 2nd to the10th post-fire year, was included in this helminthoecological study...
May 10, 2018: Parasitology Research
Andrea Galmozzi, Christopher G Parker, Bernard P Kok, Benjamin F Cravatt, Enrique Saez
Defects in adipocyte function associated with obesity drive the development of systemic insulin resistance and type 2 diabetes. Agents that correct obesity-linked adipocyte dysfunction serve as useful insulin sensitizers in humans, as is exemplified by the thiazolidinediones (TZDs). We have developed a new platform that integrates advanced chemoproteomics with phenotypic screening to identify small molecules that promote differentiation and lipid storage in adipocytes, and, in tandem, their molecular target(s)...
2018: Methods in Molecular Biology
Huong T H Nguyen, Prokopis C Andrikopoulos, Lubomír Rulíšek, Christopher J Shaffer, František Tureček
We report a gas-phase UV photodissociation study investigating non-covalent interactions between neutral hydrophobic pentapeptides and peptide ions incorporating a diazirine-tagged photoleucine residue. Phenylalanine (Phe) and proline (Pro) were chosen as the conformation-affecting residues that were incorporated into a small library of neutral pentapeptides. Gas-phase ion-molecule complexes of these peptides with photo-labeled pentapeptides were subjected to photodissociation. Selective photocleavage of the diazirine ring at 355 nm formed short-lived carbene intermediates that underwent cross-linking by insertion into H-X bonds of the target peptide...
May 7, 2018: Journal of the American Society for Mass Spectrometry
Esther Sánchez-Tirado, Araceli González-Cortés, Paloma Yáñez-Sedeño, José M Pingarrón
This paper reports the development and performance of an electrochemical immunosensor using magnetic multiwalled carbon nanotubes (m-MWCNTs) as nanocarrier tags for the determination of human fetuin A (HFA), a relevant biomarker of obesity, insulin resistance, and type-2 diabetes as well as for pancreatic and liver cancers and inflammatory processes. Screen-printed carbon electrodes were grafted with p-aminobezoic acid and streptavidin was covalently immobilized on the electrode surface. A biotinylated capture antibody was immobilized through streptavidin-biotin interaction and a sandwich assay configuration was implemented using m-MWCNTs conjugated with HRP and anti-HFA antibodies as the detection label...
April 30, 2018: Biosensors & Bioelectronics
Alexander Chamessian, Michael Young, Yawar Qadri, Temugin Berta, Ru-Rong Ji, Thomas Van de Ven
The spinal dorsal horn (SDH) is comprised of distinct neuronal populations that process different somatosensory modalities. Somatostatin (SST)-expressing interneurons in the SDH have been implicated specifically in mediating mechanical pain. Identifying the transcriptomic profile of SST neurons could elucidate the unique genetic features of this population and enable selective analgesic targeting. To that end, we combined the Isolation of Nuclei Tagged in Specific Cell Types (INTACT) method and Fluorescence Activated Nuclei Sorting (FANS) to capture tagged SST nuclei in the SDH of adult male mice...
May 1, 2018: Scientific Reports
Fangxu Xing, Jerry L Prince, Maureen Stone, Timothy G Reese, Nazem Atassi, Van J Wedeen, Georges El Fakhri, Jonghye Woo
Amyotrophic Lateral Sclerosis (ALS) is a neurological disease that causes death of neurons controlling muscle movements. Loss of speech and swallowing functions is a major impact due to degeneration of the tongue muscles. In speech studies using magnetic resonance (MR) techniques, diffusion tensor imaging (DTI) is used to capture internal tongue muscle fiber structures in three-dimensions (3D) in a non-invasive manner. Tagged magnetic resonance images (tMRI) are used to record tongue motion during speech. In this work, we aim to combine information obtained with both MR imaging techniques to compare the functionality characteristics of the tongue between normal and ALS subjects...
February 2018: Proceedings of SPIE
Louisa O'Neill, James Harrison, Henry Chubb, John Whitaker, Rahul K Mukherjee, Lars Ølgaard Bloch, Niels Peter Andersen, Høgni Dam, Henrik K Jensen, Steven Niederer, Matthew Wright, Mark O'Neill, Steven E Williams
Aims: Conducting gaps in lesion sets are a major reason for failure of ablation procedures. Voltage mapping and pace-capture have been proposed for intra-procedural identification of gaps. We aimed to compare gap size measured acutely and chronically post-ablation to macroscopic gap size in a porcine model. Methods and results: Intercaval linear ablation was performed in eight Göttingen minipigs with a deliberate gap of ∼5 mm left in the ablation line. Gap size was measured by interpolating ablation contact force values between ablation tags and thresholding at a low force cut-off of 5 g...
April 26, 2018: Europace: European Pacing, Arrhythmias, and Cardiac Electrophysiology
Romaric Bonnet, Carole Farre, Lionel Valera, Ludivine Vossier, Fanny Léon, Typhaine Dagland, Agnès Pouzet, Nicole Jaffrézic-Renault, Jeannette Fareh, Chantal Fournier-Wirth, Carole Chaix
A nanoparticle-based electrochemical sandwich immunoassay was developed for bacteria detection in platelet concentrates. For the assay, magnetic beads were functionalized with antibodies to allow the specific capture of bacteria from the complex matrix, and innovative methylene blue-DNA/nanoparticle assemblies provided the electrochemical response for amplified detection. This nanoparticular system was designed as a temperature-sensitive nano-tool for electrochemical detection. First, oligonucleotide-functionalized nanoparticles were obtained by direct synthesis of the DNA strands on the nanoparticle surface using an automated oligonucleotide synthesizer...
April 24, 2018: Analyst
Jia C Wang, Madison Bolger-Munro, Michael R Gold
B cells that bind to membrane-bound antigens (e.g., on the surface of an antigen-presenting cell) form an immune synapse, a specialized cellular structure that optimizes B-cell receptor (BCR) signaling and BCR-mediated antigen acquisition. Both the remodeling of the actin cytoskeleton and the reorientation of the microtubule network towards the antigen contact site are essential for immune synapse formation. Remodeling of the actin cytoskeleton into a dense peripheral ring of F-actin is accompanied by polarization of the microtubule-organizing center towards the immune synapse...
April 9, 2018: Journal of Visualized Experiments: JoVE
Leann Lerie Matta, Saranraj Karuppuswami, Premjeet Chahal, Evangelyn C Alocilja
Rapid detection techniques of pathogenic bacteria in the liquid food supply chain are of significant research interest due to their pivotal role in preventing foodborne outbreaks, and in maintaining high standards of public health and safety. Milk and dairy products are of particular interest due to their widespread consumption across the globe. In this paper, a biosensor for detecting pathogenic bacteria in milk using dextrin-capped gold nanoparticles (d-AuNP) as labels decoded at microwave frequencies is presented...
April 11, 2018: Biosensors & Bioelectronics
J R Ellis, G J Burt, G Grilli, S R McCully Phillips, T L Catchpole, D L Maxwell
Data on the vigour and at-vessel mortality (AVM) of 6798 skates (comprising Raja clavata n = 6295; R. brachyura n = 208; R. undulata n = 185, R. montagui n = 98 and R. microocellata n = 12) captured by commercial fishing vessels in the inshore waters of the southern North Sea and English Channel were recorded. AVM in longline fisheries averaged 0·44% across five vessels (0-1·47%), although skates were usually unhooked manually and did not usually pass through a bait-stripper. AVM in otter trawls averaged 0·76% (0-2·35%), from four vessels fishing with tow durations of <1·5 h (southern North Sea) or 1-4 h (English Channel)...
April 19, 2018: Journal of Fish Biology
Linglin Xie, Xia Yang, Yi He, Ruo Yuan, Yaqin Chai
In this work, we adopted polyacrylamide gel contained the zinc finger peptide(PZF)as a "Lock" of Raman signal and zinc ions (Zn2+) as a sensitive "key" which was converted from target-captured ZnO NPs to achieve the ultrasensitive detection of prostate-specific antigen (PSA). Owing to the "lock" effect from PZF, the SERS tag toluidine blue (TB) connected on AgNPs-coating silica wafer was sheltered leading to low Raman response. Meanwhile, target PSA can specifically connect with antibody 2 coupled ZnO nanocomplexes (ZnO@Au@Ab2) and antibody 1 coupled magnetic (CoFe2O4@Au@Ab1) through sandwich immunoassay...
April 16, 2018: ACS Applied Materials & Interfaces
Asako Isogawa, Robert P Fuchs, Shingo Fujii
The goal of present paper is to develop a reliable DNA-based method for isolation of protein complexes bound to DNA (Isolation of DNA Associated Proteins: IDAP). We describe a robust and versatile procedure to pull-down chromatinized DNA sequences-of-interest by formation of a triple helix between a sequence tag present in the DNA and a complementary triple helix forming oligonucleotide (TFO) coupled to a desthiobiotin residue. Following optimization to insure efficient recovery of native plasmids via TFO probe in vitro, the procedure is shown to work under various experimental situations...
April 12, 2018: Scientific Reports
Yong Gu, Zhiyuan Zhao, Dongmei Miao, Hilary High, Tao Yang, Liping Yu
Pinpointing islet autoantibodies associated with type 1 diabetes (T1D) leads the way to project and deter this disease in the general population. A novel ECL assay is a nonradioactive fluid phase assay for islet autoantibodies with higher sensitivity and specificity than the current 'gold' standard radio-binding assay (RBA). ECL assays can more precisely define the onset of presymptomatic T1D by distinguishing the high-risk, high-affinity autoantibodies from the low-risk, low-affinity autoantibodies generated in RBAs, and conventional enzyme-linked immunosorbent assays (ELISA)...
March 23, 2018: Journal of Visualized Experiments: JoVE
Kaizhu Zeng, Qian Li, Jing Wang, Guowei Yin, Yajun Zhang, Chaoni Xiao, Taiping Fan, Xinfeng Zhao, Xiaohui Zheng
Protein immobilization techniques play an important role in the development of assays for disease diagnosis and drug discovery. However, many of these approaches are not applicable to transmembrane proteins. G protein-coupled receptors (GPCRs) are the largest protein superfamily encoded by the human genome and are targeted by a quarter of all prescription drugs. GPCRs are highly dynamic and sensitive to changes in the ambient environment, and current immobilization methodologies are not suitable for GPCRs. We used haloalkane dehalogenase (Halo) as an immobilization tag fused to the β2 -adrenoceptor (β2 -AR), angiotensin II type 1 (AT1 ) and angiotensin II type 2 (AT2 ) receptors...
January 14, 2018: Chemical Science
Rina Komatsu, Tamayo Yamaguchi, Naohiro Kobayashi, Yoshihiro Ozeki, Kaori Sakurai
Sortin1 is an inhibitor of vesicular biogenesis and transport, which is shared among eukaryotes and plants with an unknown mode of action. Toward exploration of its target proteins, we developed alkyne as well as biotin conjugated photoaffinity probes derived from Sortin1. Due to the presence of phenylketone moiety, Sortin1 was anticipated to serve as a photoreactive group in a similar manner to a commonly used photoreactive group, benzophenone. The core structure based on 5-oxo-1,4-dihydroindenopyridine was constructed in one step using three-component Hantzsch dihydropyridine synthesis...
March 23, 2018: Bioorganic & Medicinal Chemistry Letters
Alexandra Gros, Szu-Han Wang
Decline in cognitive functions, including hippocampus-dependent spatial memory, is commonly observed at a later stage of aging (e.g., >20 months old in rodents) and typically studied after a discrete learning event. How normal aging, particularly at an early stage, affects the modulatory aspect of memory persistence is underinvestigated. Previous studies in young animals show that weak, fading memories can last longer if a modulating event, such as spatial novelty, is introduced around memory encoding. This is known as behavioral tagging and capture (BTC)...
March 10, 2018: Neurobiology of Aging
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