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https://www.readbyqxmd.com/read/29156758/development-of-novel-long-noncoding-rna-malat1-near-infrared-optical-probes-for-in-vivo-tumour-imaging
#1
Meng-Jie Dong, Cai-Qin Wang, Guo-Lin Wang, Yue-Hong Wang, Zhen-Feng Liu
With the advent of next-generation sequencing technology, there is rapidly increasing interest in long noncoding RNAs (lncRNAs). The objectives of this study were to develop a novel lncRNA MALAT1 near-infrared optical probe, to evaluate the characteristics of this optical imaging probe in vitro and to determine whether it can be used for imaging MALAT1 expression in malignant tumours in vivo. Conjugation of Cy5.5 to MALAT1 ASO was accomplished using standard NHS (N-hydroxysuccinimide) ester procedures, and the labelled MALAT1 ASO was purified with a Glen-Pak DNA Purification Cartridge and reversed-phase high performance liquid chromatography (HPLC)...
October 17, 2017: Oncotarget
https://www.readbyqxmd.com/read/29156627/multichannel-discriminative-detection-of-explosive-vapors-with-an-array-of-nanofibrous-membranes-loaded-with-quantum-dots
#2
Zhaofeng Wu, Haiming Duan, Zhijun Li, Jixi Guo, Furu Zhong, Yali Cao, Dianzeng Jia
The multichannel fluorescent sensor array based on nanofibrous membranes loaded with ZnS quantum dots (QDs) was created and demonstrated for the discriminative detection of explosives. The synergistic effect of the high surface-to-volume ratio of QDs, the good permeability of nanofibrous membranes and the differential response introduced by surface ligands was played by constructing the sensing array using nanofibrous membranes loaded with ZnS QDs featuring several surface ligands. Interestingly, although the fluorescence quenching of the nanofibrous membranes is not linearly related to the exposure time, the fingerprint of each explosive at different times is very similar in shape, and the fingerprints of the three explosives show different shapes...
November 20, 2017: Sensors
https://www.readbyqxmd.com/read/29156557/the-invasive-region-of-glioblastoma-defined-by-5ala-guided-surgery-has-an-altered-cancer-stem-cell-marker-profile-compared-to-central-tumour
#3
Stuart J Smith, Mohammed Diksin, Saachi Chhaya, Shwetha Sairam, Maria A Estevez-Cebrero, Ruman Rahman
Glioblastoma, a WHO grade IV astrocytoma, is a highly aggressive and heterogeneous tumour that infiltrates deeply into surrounding brain parenchyma, making complete surgical resection impossible. Despite chemo-radiotherapy, the residual cell population within brain parenchyma post-surgery causes inevitable recurrence. Previously, the tumour core has been the focus of research and the basis for targeted therapeutic regimes, which have failed to improve survival in clinical trials. Here, we focus on the invasive margin as defined by the region with 5-aminolevulinic acid (5ALA) (GliolanTM) fluorescence at surgery beyond the T1 enhancing region on magnetic resonance imaging (MRI)...
November 18, 2017: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/29155795/in-vivo-imaging-of-cx3cr1gfp-gfp-reporter-mice-with-spectral-domain-optical-coherence-tomography-and-scanning-laser-ophthalmoscopy
#4
Despina Kokona, Joël Jovanovic, Andreas Ebneter, Martin S Zinkernagel
Spectral domain optical coherence tomography (SD-OCT) and scanning laser ophthalmoscopy (SLO) are extensively used in experimental ophthalmology. In the present protocol, mice expressing green fluorescent protein (gfp) under the promoter of Cx3cr1 (BALB/c-Cx3cr1(gfp/gfp)) were used to image microglia cells in vivo in the retina. Microglia are resident macrophages of the retina and have been implicated in several retinal diseases(1)(,)(2)(,)(3)(,)(4)(,)(5)(,)(6). This protocol provides a detailed approach for generation of retinal B-scans, with SD-OCT, and imaging of microglia cell distribution in Cx3cr1(gfp/gfp) mice with SLO in vivo, using an ophthalmic imaging platform system...
November 11, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/29155792/assessment-of-dictyostelium-discoideum-response-to-acute-mechanical-stimulation
#5
Yulia Artemenko, Peter N Devreotes
Chemotaxis, or migration up a gradient of a chemoattractant, is the best understood mode of directed migration. Studies using social amoeba Dictyostelium discoideum revealed that a complex signal transduction network of parallel pathways amplifies the response to chemoattractants, and leads to biased actin polymerization and protrusion of a pseudopod in the direction of a gradient. In contrast, molecular mechanisms driving other types of directed migration, for example, due to exposure to shear flow or electric fields, are not known...
November 9, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/29155766/assessment-of-blood-brain-barrier-permeability-by-intravenous-infusion-of-fitc-labeled-albumin-in-a-mouse-model-of-neurodegenerative-disease
#6
Alba Di Pardo, Salvatore Castaldo, Luca Capocci, Enrico Amico, Maglione Vittorio
Disruption of blood-brain barrier (BBB) integrity is a common feature for different neurological and neurodegenerative diseases. Although the interplay between perturbed BBB homeostasis and the pathogenesis of brain disorders needs further investigation, the development and validation of a reliable procedure to accurately detect BBB alterations may be crucial and represent a useful tool for potentially predicting disease progression and developing targeted therapeutic strategies. Here, we present an easy and efficient procedure for evaluating BBB leakage in a neurodegenerative condition like that occurring in a preclinical mouse model of Huntington disease, in which defects in the permeability of BBB are clearly detectable precociously in the disease...
November 8, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/29155761/advanced-confocal-microscopy-techniques-to-study-protein-protein-interactions-and-kinetics-at-dna-lesions
#7
Soňa Legartová, Jana Suchánková, Jana Krejčí, Alena Kovaříková, Eva Bártová
Local microirradiation with lasers represents a useful tool for studies of DNA-repair-related processes in live cells. Here, we describe a methodological approach to analyzing protein kinetics at DNA lesions over time or protein-protein interactions on locally microirradiated chromatin. We also show how to recognize individual phases of the cell cycle using the Fucci cellular system to study cell-cycle-dependent protein kinetics at DNA lesions. A methodological description of the use of two UV lasers (355 nm and 405 nm) to induce different types of DNA damage is also presented...
November 12, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/29155750/ground-state-depletion-super-resolution-imaging-in-mammalian-cells
#8
Rose E Dixon, Oscar Vivas, Karen I Hannigan, Eamonn J Dickson
Advances in fluorescent microscopy and cell biology are intimately correlated, with the enhanced ability to visualize cellular events often leading to dramatic leaps in our understanding of how cells function. The development and availability of super-resolution microscopy has considerably extended the limits of optical resolution from ~250-20 nm. Biologists are no longer limited to describing molecular interactions in terms of colocalization within a diffraction limited area, rather it is now possible to visualize the dynamic interactions of individual molecules...
November 5, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/29155738/imaging-amyloid-tissues-stained-with-luminescent-conjugated-oligothiophenes-by-hyperspectral-confocal-microscopy-and-fluorescence-lifetime-imaging
#9
Sofie Nyström, Marcus Bäck, K Peter R Nilsson, Per Hammarström
Proteins that deposit as amyloid in tissues throughout the body can be the cause or consequence of a large number of diseases. Among these we find neurodegenerative diseases such as Alzheimer's and Parkinson's disease afflicting primarily the central nervous system, and systemic amyloidosis where serum amyloid A, transthyretin and IgG light chains deposit as amyloid in liver, carpal tunnel, spleen, kidney, heart, and other peripheral tissues. Amyloid has been known and studied for more than a century, often using amyloid specific dyes such as Congo red and Thioflavin T (ThT) or Thioflavin (ThS)...
October 20, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/29155734/structural-characterization-of-mannan-cell-wall-polysaccharides-in-plants-using-pace
#10
Venkataramana R Pidatala, Amir Mahboubi, Jenny C Mortimer
Plant cell wall polysaccharides are notoriously difficult to analyze, and most methods require expensive equipment, skilled operators, and large amounts of purified material. Here, we describe a simple method for gaining detailed polysaccharide structural information, including resolution of structural isomers. For polysaccharide analysis by gel electrophoresis (PACE), plant cell wall material is hydrolyzed with glycosyl hydrolases specific to the polysaccharide of interest (e.g., mannanases for mannan). Large format polyacrylamide gels are then used to separate the released oligosaccharides, which have been fluorescently labeled...
October 16, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/29155726/visualization-and-live-imaging-of-oligodendrocyte-organelles-in-organotypic-brain-slices-using-adeno-associated-virus-and-confocal-microscopy
#11
Lauritz Hagen Kennedy, Johanne Egge Rinholm
Neurons rely on the electric insulation and trophic support of myelinating oligodendrocytes. Despite the importance of oligodendrocytes, the advanced tools currently used to study neurons, have only partly been taken on by oligodendrocyte researchers. Cell type-specific staining by viral transduction is a useful approach to study live organelle dynamics. This paper describes a protocol for visualizing oligodendrocyte mitochondria in organotypic brain slices by transduction with adeno-associated virus (AAV) carrying genes for mitochondrial targeted fluorescent proteins under the transcriptional control of the myelin basic protein promoter...
October 23, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/29155237/a-standardized-toolkit-for-genetic-engineering-of-ctg-clade-yeasts
#12
Tatiana A Defosse, Vincent Courdavault, Alix T Coste, Marc Clastre, Thomas Dugé de Bernonville, Charlotte Godon, Patrick Vandeputte, Arnaud Lanoue, Antoine Touzé, Tomas Linder, Samir Droby, Carlos A Rosa, Dominique Sanglard, Christophe d'Enfert, Jean-Philippe Bouchara, Nathalie Giglioli-Guivarc'h, Nicolas Papon
We have developed a series of synthetic constructs suitable to genetically manipulate a broad range of yeast species belonging to the fungal CTG clade. This molecular toolbox notably allows heterologous gene expression, single or dual fluorescence labeling and construction of luciferase-expressing strains for bioluminescence imaging.
November 16, 2017: Journal of Microbiological Methods
https://www.readbyqxmd.com/read/29155042/imaging-flow-cytometry-to-assess-chromosomal-abnormalities-in-chronic-lymphocytic-leukaemia
#13
Henry Hui, Kathryn A Fuller, Hun Chuah, James Liang, Hasib Siddiqi, Dejan Radeski, Wendy N Erber
Chronic Lymphocytic Leukaemia (CLL), the most common leukaemia in the Western world, has a characteristic phenotype and prognosis largely defined by the presence of cytogenetic aberrations. The gold standard for detecting these cytogenetic abnormalities is interphase fluorescence in situ hybridisation (FISH) performed on cell smears or tissue sections on glass slides. Fluorescently labelled DNA probes bind to specific chromosomal regions and the signal detected by fluorescent microscopy. Generally only 200 cells are assessed and the limit of sensitivity is 3% positive cells...
November 15, 2017: Methods: a Companion to Methods in Enzymology
https://www.readbyqxmd.com/read/29155038/a-novel-modernized-golgi-cox-stain-optimized-for-clarity-cleared-tissue
#14
Mustafa S Kassem, Sandra Y Y Fok, Kristie L Smith, Michael Kuligowski, Bernard W Balleine
BACKGROUND: High resolution neuronal information is extraordinarily useful in understanding the brain's functionality. The development of the Golgi-Cox stain allowed observation of the neuron in its entirety with unrivalled detail. Recently tissue clearing techniques, e.g., CLARITY and CUBIC, provide the potential to observe entire neuronal circuits intact within tissue and without previous restrictions with regard to section thickness. NEW METHOD: Here we describe an improved Golgi-Cox stain method, optimized for use with CLARITY and CUBIC, that stains within 48hrs and that can be used in both fresh and fixed tissue...
November 15, 2017: Journal of Neuroscience Methods
https://www.readbyqxmd.com/read/29154961/inhibition-of-cyclooxygenase-cox-2-suppresses-the-recruitment-of-endothelial-progenitor-cells-in-the-microvasculature-of-endometriotic-lesions
#15
Jeannette Rudzitis-Auth, Ruth M Nickels, Michael D Menger, Matthias W Laschke
The incorporation of endothelial progenitor cells (EPCs) into newly developing blood vessels contributes to the vascularization of endometriotic lesions. here, we analyzed whether cyclooxygenase (COX)-2 signaling regulates this vasculogenic process. Endometriotic lesions were surgically induced in irradiated FVB/N mice, which were reconstituted with bone marrow from FVB/N-TgN (Tie2/green fluorescent protein (GFP)) 287 Sato mice. The animals received β-estradiol 17-valerate once a week and were daily treated with the selective COX-2 inhibitor parecoxib (25 mg/kg) or vehicle (control) over 7 and 28 days...
November 15, 2017: American Journal of Pathology
https://www.readbyqxmd.com/read/29154543/fluorogenic-targeting-of-voltage-sensitive-dyes-to-neurons
#16
Pei Liu, Vincent Grenier, Wootack Hong, Vikram R Muller, Evan W Miller
We present a method to target voltage-sensitive fluorescent dyes to specified cells using an enzyme-catalyzed fluorogenic reaction on cell surfaces. The dye/enzyme hybrids are composed of a photoinduced electron transfer (PeT)-based fluorescent voltage indicator and a complementary enzyme expressed on the cell surface. Action of the exogenous enzyme on the dye results in fluorogenic activation of the dye, enabling fast voltage imaging in defined neurons with sensitivity surpassing those of purely genetically encoded approaches...
November 20, 2017: Journal of the American Chemical Society
https://www.readbyqxmd.com/read/29154217/a-novel-turn-on-fluorescent-probe-for-al-3-and-fe-3-in-aqueous-solution-and-its-imaging-in-living-cells
#17
Yanpeng Dai, Jiaxin Fu, Kun Yao, Qianqian Song, Kuoxi Xu, Xiaobin Pang
A quinoline-based fluorescence probe has been prepared and characterized. Probe 1 showed a selective sensing ability for Al(3+) and Fe(3+) ions through fluorescence enhancement response at 515nm when it was excited at 360nm. In the presence of Fe(3+) ion, probe 1 exhibited a detection limit of 2.10×10(-6)M. As for Al(3+), its detection limit of 3.58×10(-7)M was significantly lower than the highest limit of Al(3+) in drinking water recommended by the WHO (7.41μM), representing a rare example in reported fluorescent probe for Al(3+) ion...
November 7, 2017: Spectrochimica Acta. Part A, Molecular and Biomolecular Spectroscopy
https://www.readbyqxmd.com/read/29154213/liposomally-formulated-phospholipid-conjugated-novel-near-infrared-fluorescence-probe-for-particle-size-effect-on-cellular-uptake-and-biodistribution-in-vivo
#18
Jing Xing, Dong Liu, Gaoxin Zhou, Yuan Li, Peng Wang, Ke Hu, Ning Gu, Min Ji
Lipid based nanoparticles (LBNs) with excellent biocompatibility and versatility have received much attention from the drug delivery community recently. A detailed understanding of in vitro and vivo fate of LBNs is important for developing different types of LBNs with improved selectivity and low cytotoxicity. We developed a novel near-infrared (NIR) probe with high fluorescence, designated as DSPE-ir623 (iDSPE). Then, we prepared iDSPE-embeded liposomes (iLPs) with two different hydrodynamic sizes (∼100nm and ∼400nm) to evaluate the effect of particle size on cellular uptake and biodistribution of nanoliposomes in vivo...
November 16, 2017: Colloids and Surfaces. B, Biointerfaces
https://www.readbyqxmd.com/read/29154189/novel-theranostic-zinc-phthalocyanine-phospholipid-complex-self-assembled-nanoparticles-for-imaging-guided-targeted-photodynamic-treatment-with-controllable-ros-production-and-shape-assisted-enhanced-cellular-uptake
#19
Jinyuan Ma, Yang Li, Guihua Liu, Ai Li, Yilin Chen, Xinyi Zhou, Dengyue Chen, Zhenqing Hou, Xuan Zhu
The novel drug delivery system based on self-assembly of zinc phthalocyanine-soybean phosphatidylcholine (ZnPc-SPC) complex was developed by a co-solvent method followed by a nanoprecipitaion technique. DSPE-PEG-methotrexate (DSPE-PEG-MTX) was introduced on the surface of ZnPc-SPC self-assembled nanoparticles (ZS) to endow them with folate receptor-targeting property. NMR, XRD, FTIR, and UV-vis-NIR analysis demonstrated the weak molecular interaction between ZnPc and SPC. The ZS functionalized with DSPE-PEG-MTX (ZSPM) was successfully constructed with an average particle size of ∼170nm, a narrow size distribution, and could remain physiologically stable for at least 7days...
October 24, 2017: Colloids and Surfaces. B, Biointerfaces
https://www.readbyqxmd.com/read/29153902/robotic-buccal-mucosal-graft-ureteroplasty-for-complex-ureteral-stricture
#20
Sohrab Arora, Logan Campbell, Mouafak Tourojman, Daniel Pucheril, Lamont R Jones, Craig Rogers
OBJECTIVE: To demonstrate robot-assisted ureterolysis and buccal mucosal graft (BMG) ureteroplasty for the management of a complex, long recurrent ureteral stricture developing after ureterolysis, and also to demonstrate the use of near-infrared fluorescence (NIRF) imaging and intraoperative ureteroscopy during this procedure. METHODS: A 58-year-old man with a history of cabergoline treatment and a cardiac catheterization through the left groin presented with left flank pain and hydronephrosis...
December 2017: Urology
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