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https://www.readbyqxmd.com/read/27911496/gram-scale-chemical-synthesis-of-base-modified-ribonucleoside-5-o-triphosphates
#1
Muthian Shanmugasundaram, Annamalai Senthilvelan, Anilkumar R Kore
This unit delineates a simple, reliable, straight-forward, general, and efficient chemical method for the synthesis of modified nucleoside-5'-O-triphosphates such as 5-methylcytidine-5'-O-triphosphate (5-Me-CTP), pseudouridine-5'-O-triphosphate (pseudo-UTP), and N(1) -methylpseudouridine-5'-O-triphosphate (N(1) -methylpseudo-UTP), starting from the corresponding nucleoside. The reaction utilizes an improved protection-free "one-pot, three-step" Ludwig synthetic strategy that involves the monophosphorylation of the nucleoside with phosphorous oxychloride followed by reaction with tributylammonium pyrophosphate and subsequent hydrolysis of the resulting cyclic intermediate to furnish the corresponding ribonucleoside triphosphate (NTP) in moderate yields...
December 1, 2016: Current Protocols in Nucleic Acid Chemistry
https://www.readbyqxmd.com/read/27911492/rolling-circle-amplification-with-chemically-modified-nucleoside-triphosphates
#2
Marcel Hollenstein, Masad J Damha
Modified nucleoside triphosphates (dN*TPs) represent facile and versatile precursors for the introduction of chemical diversity into nucleic acids. While dN*TPs have been utilized in a plethora of practical applications, very little attention has been devoted to the assessment of their compatibility with isothermal amplification strategies. In this context, rolling circle amplification (RCA) is a wide-spread enzymatic replication method in which small single-stranded DNA (ssDNA) circles serve as templates in primer extension reactions yielding very long, ssDNA products...
December 1, 2016: Current Protocols in Nucleic Acid Chemistry
https://www.readbyqxmd.com/read/27911489/palliative-care-interventions-in-advanced-dementia
#3
REVIEW
Edel Murphy, Katherine Froggatt, Sheelah Connolly, Eamon O'Shea, Elizabeth L Sampson, Dympna Casey, Declan Devane
BACKGROUND: Dementia is a chronic, progressive and ultimately fatal neurodegenerative disease. Advanced dementia is characterised by profound cognitive impairment, inability to communicate verbally and complete functional dependence. Usual care of people with advanced dementia is not underpinned universally by a palliative approach. Palliative care has focused traditionally on care of people with cancer but for more than a decade, there have been increased calls worldwide to extend palliative care services to include all people with life-limiting illnesses in need of specialist care, including people with dementia...
December 2, 2016: Cochrane Database of Systematic Reviews
https://www.readbyqxmd.com/read/27911481/photoluminescence-and-thermoluminescence-of-k2-mg-so4-2-eu-and-evaluation-of-its-kinetic-parameters
#4
Archana Deshpande, N S Dhoble, S C Gedam, S J Dhoble
The K2 Mg(SO4 )2 :Eu phosphor, synthesized by a solid-state diffusion method, was studied for its photoluminescence (PL) and thermoluminescence (TL) characteristics. The X-ray diffraction (XRD) pattern of the material was matched with the standard JCPDF No. 36-1499. For PL characteristics, K2 Mg(SO4 )2 :Eu(2)(+) showed an emission peak at 474 nm when excited at 340 nm, while it showed Eu(3)(+) emission at 580 nm, and 594 nm splitting at 613 nm and 618 nm for an excitation of 396 nm wavelength due to radiative transitions from (5) D0 to (7) Fj (j = 0, 1, 2, 3)...
December 2, 2016: Luminescence: the Journal of Biological and Chemical Luminescence
https://www.readbyqxmd.com/read/27911478/modulation-of-whistle-production-related-to-training-sessions-in-bottlenose-dolphins-tursiops-truncatus-under-human-care
#5
Juliana Lopez Marulanda, Olivier Adam, Fabienne Delfour
Bottlenose dolphins are highly social cetaceans with an extensive sound production including clicks, burst-pulsed sounds, and whistles. Some whistles, known as signature whistles, are individually specific. These acoustic signatures are commonly described as being emitted in contexts of stress during forced isolation and as group cohesion calls. Interactions between humans and captive dolphins is largely based on positive reinforcement conditioning within several training/feeding sessions per day. Vocal behavior of dolphins during these interactions might vary...
November 2016: Zoo Biology
https://www.readbyqxmd.com/read/27911467/a-systematic-investigation-of-the-charging-effect-in-scanning-electron-microscopy-for-metal-nanostructures-on-insulating-substrates
#6
R Flatabø, A Coste, M M Greve
Scanning electron microscopy is perhaps the most important method for investigating and characterizing nanostructures. A well-known challenge in scanning electron microscopy is the investigation of insulating materials. As insulating materials do not provide a path to ground they accumulate charge, evident as image drift and image distortions. In previous work, we have seen that sample charging in arrays of metal nanoparticles on glass substrates leads to a shrinkage effect, resulting in a measurement error in the nanoparticle dimension of up to 15% at 10 kV and a probe current of 80 ± 10 pA...
December 2, 2016: Journal of Microscopy
https://www.readbyqxmd.com/read/27911451/influence-of-specific-intermolecular-interactions-on-the-thermal-and-dielectric-properties-of-bulk-polymers-atomistic-molecular-dynamics-simulations-of-nylon-6
#7
N V Lukasheva, D A Tolmachev, V M Nazarychev, J M Kenny, S V Lyulin
Specific intermolecular interactions, in particular H-bonding, have a strong influence on the structural, thermal and relaxation characteristics of polymers. We report here the results of molecular dynamics simulations of Nylon 6 which provides an excellent example for the investigation of such an influence. To demonstrate the effect of proper accounting for H-bonding on bulk polymer properties, the AMBER99sb force field is used with two different parametrization approaches leading to two different sets of partial atomic charges...
December 2, 2016: Soft Matter
https://www.readbyqxmd.com/read/27911445/long-term-outcomes-of-punch-punctoplasty-with-kelly-punch-and-review-of-literature
#8
E S Wong, E Y Li, H K Yuen
PurposeTo report long-term outcomes of punch punctoplasty utilizing the Kelly punch and to compare the results with other described methods of punctoplasty in literature.Patients and methodsA retrospective, non-comparative interventional case series of patients who underwent punch punctoplasty at the Hong Kong Eye Hospital over an 8-year period. A standard Kelly Descemet's membrane punch was utilized for punctal enlargement in all cases. Patient records and their operative records were reviewed. Anatomical success was defined by well-patent puncta on follow-up...
December 2, 2016: Eye
https://www.readbyqxmd.com/read/27911435/detecting-biomarkers-of-secondhand-marijuana-smoke-in-young-children
#9
Karen M Wilson, Michelle R Torok, Binnian Wei, Lanqing Wang, Michelle Robinson, Connie S Sosnoff, Benjamin C Blount
BACKGROUND: The impact of secondhand marijuana smoke exposure on children is unknown. New methods allow detection of secondhand marijuana smoke in children. METHODS: We studied children ages 1 month to 2 years hospitalized with bronchiolitis in Colorado from 2013-2015. Parents completed a survey, and urine samples were analyzed for cotinine using LC/MS/MS (LOD 0.03 ng/ml) and marijuana metabolites including COOH-THC (LOD 0.015 ng/ml). RESULTS: A total of 43 subjects had urine samples available for analysis...
December 2, 2016: Pediatric Research
https://www.readbyqxmd.com/read/27911427/-modern-approach-to-early-diagnosis-and-pathogenetic-treatment-of-diabetic-retinopathy
#10
I V Vorob'eva
AIM: To improve the approach to pathogenetic treatment of diabetic retinopathy (DR) through early diagnosis and a new method for predicting disease progression. MATERIAL AND METHODS: The study enrolled 330 type 2 diabetes patients with DR (660 eyes), of whom women constituted 64.6%, men - 35.4%. The mean patient age was 62.3±2.3 years. Three groups were formed: the controls - 30 healthy volunteers (60 eyes) and 30 type 2 diabetes patients without ocular involvement (DR 0, 60 eyes); group 1 - 30 type 2 diabetes patients with DR I but no diabetic macular edema (DR I without DME, 60 eyes) that were treated with calcium dobesilate; group 2 - 240 type 2 diabetes patients, who had diabetic retinopathy of different stages (DR I, II, or III with DME, 480 eyes) and received laser retinal photocoagulation (LRP)...
2016: Vestnik Oftalmologii
https://www.readbyqxmd.com/read/27911424/-effectiveness-of-soft-contact-lenses-in-severe-keratoconjunctivitis-sicca-treated-with-local-immunosuppressive-therapy
#11
T N Safonova, O V Gladkova, V I Boev, A A Fedorov
AIM: To study the effectiveness of soft contact lenses in severe keratoconjunctivitis sicca (KCS) treated with local immunosuppressive therapy. MATERIAL AND METHODS: We have examined 41 patients (60 eyes) with severe KCS, including 3 men (4 eyes) and 38 women (56 eyes) aged from 27 to 76 years. Group 1 consisted of 22 persons (30 eyes) who were prescribed cyclosporine А 0.05% instillations 2 times daily, artificial tears, and soft contact lenses. Group 2 consisted of 19 patients (30 eyes) who received instillations of cyclosporine А 0...
2016: Vestnik Oftalmologii
https://www.readbyqxmd.com/read/27911419/methods-of-soil-resampling-to-monitor-changes-in-the-chemical-concentrations-of-forest-soils
#12
Gregory B Lawrence, Ivan J Fernandez, Paul W Hazlett, Scott W Bailey, Donald S Ross, Thomas R Villars, Angelica Quintana, Rock Ouimet, Michael R McHale, Chris E Johnson, Russell D Briggs, Robert A Colter, Jason Siemion, Olivia L Bartlett, Olga Vargas, Michael R Antidormi, Mary M Koppers
Recent soils research has shown that important chemical soil characteristics can change in less than a decade, often the result of broad environmental changes. Repeated sampling to monitor these changes in forest soils is a relatively new practice that is not well documented in the literature and has only recently been broadly embraced by the scientific community. The objective of this protocol is therefore to synthesize the latest information on methods of soil resampling in a format that can be used to design and implement a soil monitoring program...
November 25, 2016: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/27911418/synthesis-and-characterization-of-fe-doped-aluminosilicate-nanotubes-with-enhanced-electron-conductive-properties
#13
Ehsan Shafia, Serena Esposito, Elnaz Bahadori, Marco Armandi, Maela Manzoli, Barbara Bonelli
The goal of the protocol is to synthesize Fe-doped aluminosilicate nanotubes of the imogolite type with the formula (OH)3Al2-xFexO3SiOH. Doping with Fe aims at lowering the band gap of imogolite, an insulator with the chemical formula (OH)3Al2O3SiOH, and at modifying its adsorption properties towards azo-dyes, an important class of organic pollutants of both wastewater and groundwater. Fe-doped nanotubes are obtained in two ways: by direct synthesis, where FeCl3 is added to an aqueous mixture of the Si and Al precursors, and by post-synthesis loading, where preformed nanotubes are put in contact with a FeCl3•6H2O aqueous solution...
November 15, 2016: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/27911414/isolation-and-culture-of-primary-endothelial-cells-from-canine-arteries-and-veins
#14
Loes A Oosterhoff, Hedwig S Kruitwagen, Bart Spee, Frank G van Steenbeek
Cardiovascular disease is studied in both human and veterinary medicine. Endothelial cells have been used extensively as an in vitro model to study vasculogenesis, (tumor) angiogenesis, and atherosclerosis. The current standard for in vitro research on human endothelial cells (ECs) is the use of Human Umbilical Vein Endothelial Cells (HUVECs) and Human Umbilical Artery Endothelial Cells (HUAECs). For canine endothelial research, only one cell line (CnAOEC) is available, which is derived from canine aortic endothelium...
November 18, 2016: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/27911413/fabricating-a-uv-vis-and-raman-spectroscopy-immunoassay-platform
#15
Cynthia Hanson, Nathan D Israelsen, Michael Sieverts, Elizabeth Vargis
Immunoassays are used to detect proteins based on the presence of associated antibodies. Because of their extensive use in research and clinical settings, a large infrastructure of immunoassay instruments and materials can be found. For example, 96- and 384-well polystyrene plates are available commercially and have a standard design to accommodate ultraviolet-visible (UV-Vis) spectroscopy machines from various manufacturers. In addition, a wide variety of immunoglobulins, detection tags, and blocking agents for customized immunoassay designs such as enzyme-linked immunosorbent assays (ELISA) are available...
November 10, 2016: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/27911412/reproducible-arterial-denudation-injury-by-infrarenal-abdominal-aortic-clamping-in-a-murine-model
#16
Aditya S Shirali, Austin I McDonald, Julia J Mack, M Luisa Iruela-Arispe
Percutaneous vascular interventions uniformly result in arterial denudation injuries that subsequently lead to thrombosis and restenosis. These complications can be attributed to impairments in re-endothelialization within the wound margins. Yet, the cellular and molecular mechanisms of re-endothelialization remain to be defined. While several animal models to study re-endothelialization after arterial denudation are available, few are performed in the mouse because of surgical limitations. This undermines the opportunity to exploit transgenic mouse lines and investigate the contribution of specific genes to the process of re-endothelialization...
November 24, 2016: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/27911411/preparation-and-in-vivo-use-of-an-activity-based-probe-for-n-acylethanolamine-acid-amidase
#17
Elisa Romeo, Silvia Pontis, Stefano Ponzano, Fabiola Bonezzi, Marco Migliore, Simona Di Martino, Maria Summa, Daniele Piomelli
Activity-based protein profiling (ABPP) is a method for the identification of an enzyme of interest in a complex proteome through the use of a chemical probe that targets the enzyme's active sites. A reporter tag introduced into the probe allows for the detection of the labeled enzyme by in-gel fluorescence scanning, protein blot, fluorescence microscopy, or liquid chromatography-mass spectrometry. Here, we describe the preparation and use of the compound ARN14686, a click chemistry activity-based probe (CC-ABP) that selectively recognizes the enzyme N-acylethanolamine acid amidase (NAAA)...
November 23, 2016: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/27911408/defining-substrate-specificities-for-lipase-and-phospholipase-candidates
#18
Diana X Sahonero-Canavesi, Maritza Zavaleta-Pastor, Lourdes Martínez-Aguilar, Isabel M López-Lara, Otto Geiger
Microorganisms produce a wide spectrum of (phospho)lipases that are secreted in order to make external substrates available for the organism. Alternatively, other (phospho)lipases may be physically associated with the producing organism causing a turnover of intrinsic lipids and frequently giving rise to a remodeling of the cellular membranes. Although potential (phospho)lipases can be predicted with a number of algorithms when the gene/protein sequence is available, experimental proof of the enzyme activities, substrate specificities, and potential physiological functions has frequently not been obtained...
November 23, 2016: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/27911407/preparation-of-cd4-t-cells-for-analysis-of-gd3-and-gd2-ganglioside-membrane-expression-by-microscopy
#19
Tania M Villanueva-Cabello, Iván Martinez-Duncker
The methods described herein for activation of naïve CD4(+) T cells in suspension and their adherence in coverslips for confocal microscopy analysis allow the spatial localization and visualization of gangliosides involved in CD4(+) T cell activation, that complement expression profiling experiments such as flow cytometry, western blotting or real-time PCR. The quantification of ganglioside expression through flow cytometry and their cellular localization through microscopy can be obtained by the use of anti-ganglioside antibodies with high affinity and specificity...
November 8, 2016: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/27911406/ultrasound-guided-botulinum-toxin-a-injections-a-method-of-treating-sialorrhea
#20
Pierangelo Barbero, Marco Busso, Carlo Alberto Artusi, Stefania De Mercanti, Marco Tinivella, Andrea Veltri, Luca Durelli, Marinella Clerico
Neurological diseases can be complicated by sialorrhea, an excessive flow of saliva. Patients suffering from moderate to severe sialorrhea have an impaired quality of life, often worsened by correlated complications such as aspiration pneumonia, oral infections, dental caries, and maceration of the skin. Diverse therapeutic approaches have been proposed for the treatment of sialorrhea, including surgery and the use of anticholinergic agents, with limited results and the possible occurrence of serious adverse events...
November 9, 2016: Journal of Visualized Experiments: JoVE
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