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https://www.readbyqxmd.com/read/28811218/wnt-signaling-positively-regulates-endothelial-cell-fate-specification-in-the-fli1a-positive-progenitor-population-via-lef1
#1
Kathleen Hübner, Kathrin S Grassme, Jyoti Rao, Nina K Wenke, Cordula L Zimmer, Laura Korte, Katja Mu Ller, Saulius Sumanas, Boris Greber, Wiebke Herzog
During vertebrate embryogenesis, vascular endothelial cells (ECs) and primitive erythrocytes become specified within close proximity in the posterior lateral plate mesoderm (LPM) from a common progenitor. However, the signaling cascades regulating the specification into either lineage remain largely elusive. Here, we analyze the contribution of β-catenin dependent Wnt signaling to EC and erythrocyte specification during zebrafish embryogenesis. We generated novel β-catenin dependent Wnt signaling reporters which, by using destabilized fluorophores (Venus-Pest, dGFP), specifically allow us to detect Wnt signaling responses in narrow time windows as well as in spatially restricted domains, defined by Cre recombinase expression (Tg(axin2BAC:Venus-Pest)(mu288); Tg(14TCF:loxP-STOP-loxP-dGFP)(mu202))...
August 12, 2017: Developmental Biology
https://www.readbyqxmd.com/read/28811025/simultaneous-detection-of-two-hepatocellar-carcinoma-related-micrornas-using-a-clever-single-labeled-fluorescent-probe
#2
Rong Liao, Shiyu Li, Huan Wang, Chunyan Chen, Xiaoming Chen, Changqun Cai
The simultaneous detection of two important microRNAs (miRNAs) can potentially evaluate pathological states. The simultaneous detection of miRNA-26a and miRNA-122 is proposed in this study using a cleverly designed single-labeled, dual-functional fluorescent probe with a 2-aminopurine as a fluorophore, which is a G-quadruplex single-stranded DNA. The probe can partly complement cDNA-1 and cDNA-2. cDNA-1 and cDNA-2 are complementary strands of miRNA-26a and miRNA-122, respectively. When the target miRNAs are added simultaneously, these cDNA (cDNA-1 and cDNA-2) can be competed off from the cDNA∖probe duplex to form a cDNA∖RNA heteroduplex...
August 29, 2017: Analytica Chimica Acta
https://www.readbyqxmd.com/read/28810739/a-hexa-rhodium-metallopeptide-catalyst-for-site-specific-functionalization-of-natural-antibodies
#3
Jun Ohata, Zachary T Ball
Preparation of antibody-drug conjugates (ADCs), an emerging class of targeted biological hybrid agents, necessitates precise control of conjugation reactivity. Antibodies have complex multi-stranded architectures, and specific modification of natu-ral antibodies has proven quite challenging. Here, we demonstrate that cooperative activity of a multi-metallic metallopeptide enables efficient site-specific antibody functionalization, based on molecular recognition of the constant Fc region. This interplay of multiple metal centers enables introduction of an orthogonal alkyne handle into monoclonal or polyclonal antibodies from different species...
August 16, 2017: Journal of the American Chemical Society
https://www.readbyqxmd.com/read/28807148/acetylcholine-induces-intracellular-ca-2-oscillations-and-nitric-oxide-release-in-mouse-brain-endothelial-cells
#4
Estella Zuccolo, Dmitry Lim, Dlzar Ali Kheder, Angelica Perna, Paolo Catarsi, Laura Botta, Vittorio Rosti, Laura Riboni, Giulio Sancini, Franco Tanzi, Egidio D'Angelo, Germano Guerra, Francesco Moccia
Basal forebrain neurons increase cortical blood flow by releasing acetylcholine (Ach), which stimulates endothelial cells (ECs) to produce the vasodilating gasotransmitter, nitric oxide (NO). Surprisingly, the mechanism whereby Ach induces NO synthesis in brain microvascular ECs is unknown. An increase in intracellular Ca(2+) concentration recruits a multitude of endothelial Ca(2+)-dependent pathways, such as Ca(2+)/calmodulin endothelial NO synthase (eNOS). The present investigation sought to investigate the role of intracellular Ca(2+) signaling in Ach-induced NO production in bEND5 cells, an established model of mouse brain microvascular ECs, by conventional imaging of cells loaded with the Ca(2+)-sensitive dye, Fura-2/AM, and the NO-sensitive fluorophore, DAF-DM diacetate...
September 2017: Cell Calcium
https://www.readbyqxmd.com/read/28807010/angiogenic-potency-evaluation-of-cell-therapy-candidates-by-a-novel-application-of-the-in-vitro-aortic-ring-assay
#5
Farwah Iqbal, Peter Szaraz, Matthew Librach, Andrée Gauthier-Fisher, Clifford L Librach
BACKGROUND: Due to limitations of current angiogenesis assays, we aimed to develop a novel application of the rat aortic ring assay to assess the angiogenic potential of mesenchymal stromal cells (MSCs). First-trimester human umbilical cord-derived perivascular cells (FTM HUCPVCs) have multipotent characteristics and previously demonstrated angiogenic potential. We compared the effect of this young source of MSCs and adult bone marrow stromal cells (BMSCs) on ex vivo aortic endothelial network formation...
August 14, 2017: Stem Cell Research & Therapy
https://www.readbyqxmd.com/read/28806521/determination-of-caspase-3-activity-and-its-inhibition-constant-by-combination-of-fluorescence-correlation-spectroscopy-with-a-microwell-chip
#6
Di Su, Xiaocai Hu, Chaoqing Dong, Jicun Ren
Caspase-3 is a key enzyme executing apoptosis during ontogenesis and homeostasis of multicellular organisms, and is a very important and potential drug target in treatment of apoptosis disturbance. So far, no commercial drugs for caspase-3 are available, and it is urgently necessitated to develop an effective method for caspase-3 activity assay and its inhibitor screening. In this paper, we propose a new method for determination of caspase-3 activity and its inhibition constant by combining single molecule fluorescence correlation spectroscopy (FCS) with a microwell chip...
August 14, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/28806519/pluronic-p85-f68-micelles-of-baicalein-could-interfere-with-mitochondria-to-overcome-mrp2-mediated-efflux-and-offer-improved-anti-parkinsonian-activity
#7
Tongkai Chen, Ye Li, Chuwen Li, Xiang Yi, Ruibing Wang, Simon Ming Yuen Lee, Ying Zheng
Overexpression of the drug efflux transporter multidrug resistance-associated protein 2 (MRP2) in the gastrointestinal tract and blood-brain barrier compromises the oral delivery of drugs to the circulation system and brain in the treatment of Parkinson's disease (PD). In this study, we aim to develop small-sized Pluronic P85/F68 micelles loaded with baicalein (B-MCs) to overcome MRP2-mediated efflux and to investigate related mechanism, as well as the anti-Parkinsonian efficacy. Spherical and sustained-release B-MCs have a mean particle size of 40...
August 14, 2017: Molecular Pharmaceutics
https://www.readbyqxmd.com/read/28806473/flavylium-polymethine-fluorophores-for-imaging-in-the-near-and-shortwave-infrared
#8
Emily D Cosco, Justin R Caram, Oliver T Bruns, Daniel Franke, Rachael A Day, Erik P Farr, Moungi G Bawendi, Ellen May Sletten
Bright fluorophores in the near infrared and shortwave infrared (SWIR) are essential for optical imaging in vivo. Herein, we utilize a 7-dimethylamino flavylium heterocycle to construct a panel of novel red-shifted polymethine dyes, with emission from 680 to 1045 nm. Photophysical characterization reveals that the 1- and 3-methine dyes display enhanced photostabilities and the 5- and 7-methine dyes exhibit exceptional brightness for their respective spectral regions. A micelle formulation of the 7-methine facilitates SWIR imaging in mice...
August 14, 2017: Angewandte Chemie
https://www.readbyqxmd.com/read/28806064/conditional-displacement-hybridization-assay-for-multiple-snp-phasing
#9
Tsz Wing Fan, Henson L Lee Yu, I-Ming Hsing
The two chromosomal copies of the human genome are highly polymorphic, and the allelic content on each strand can dictate a person's biological outcomes. While many of the current diagnostic tools are able to detect the presence of multiple mutations at the same time, most cannot determine the phase of these mutations unless long-range PCR or sequencing techniques are used or if templates are compartmentalized into single copies prior to amplification. Here, an enzyme-coupled hybridization assay, named Conditional Displacement Hybridization Assay (CDHA), is described for the concurrent and rapid determination of the presence and phase of SNP variants...
August 14, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/28803543/bodipy-based-dye-for-no-wash-live-cell-staining-and-imaging
#10
Alexey A Pakhomov, Igor E Deyev, Natalia M Ratnikova, Stepan P Chumakov, Veronika B Mironiuk, Yuriy N Kononevich, Aziz M Muzafarov, Vladimir I Martynov
In nonpolar solvents, hydrophobic organic fluorophores often show bright fluorescence, whereas in polar media, they usually suffer from aggregation-caused quenching (ACQ). Here, we harnessed this solvatochromic behavior of a 1,3,5,7-tetramethyl-BODIPY derivative for cell staining and applied it to live-cell imaging and flow cytometry. As opposed to commercially available dyes, this BODIPY derivative showed excellent contrast immediately after staining and did not require any wash-off.
August 1, 2017: BioTechniques
https://www.readbyqxmd.com/read/28802172/interaction-of-phenazinium-based-photosensitizers-with-the-n-and-b-isoforms-of-human-serum-albumin-effect-of-methyl-substitution
#11
Swagata Sen, Riya Sett, Bijan K Paul, Nikhil Guchhait
The present work is focused on exploring the interaction of two phenazinium-based biological photosensitizers, phenosafranin (PSF) and safranin-O (SO), with human serum albumin (HSA), with particular emphasis on the physiologically significant NB conformational transition of the protein on the dye:HSA interaction. In addition, the presence of methyl substitution on the planar phenazinium ring in SO paves way for looking into the effect of simple chemical manipulation (that is, methyl substitution on the dye nucleus) on the dye:protein interaction behavior as a function of various (pH-induced) isoforms of HSA...
August 3, 2017: Journal of Photochemistry and Photobiology. B, Biology
https://www.readbyqxmd.com/read/28800466/monitoring-and-modulating-ion-traffic-in-hybrid-lipid-polymer-vesicles
#12
Walter F Paxton, Patrick T McAninch, Komandoor E Achyuthan, Sun Hae Ra Shin, Haley L Monteith
Controlling the traffic of molecules and ions across membranes is a critical feature in a number of biologically relevant processes and highly desirable for the development of technologies based on membrane materials. In this paper, ion transport behavior of hybrid lipid/polymer membranes was studied in the absence and presence of ion transfer agents. A pH-sensitive fluorophore was used to investigate ion (H(+)/OH(-)) permeability across hybrid lipid/polymer membranes as a function of the fraction of amphiphilic block copolymer...
August 1, 2017: Colloids and Surfaces. B, Biointerfaces
https://www.readbyqxmd.com/read/28799205/effects-of-fluorophore-attachment-on-protein-conformation-and-dynamics-studied-by-spfret-and-nmr
#13
Carolina Sanchez, Lena Voith-von-Voithenberg, Lisa Warner, Don C Lamb, Michael Sattler
Fluorescence-based techniques are widely used to study biomolecular conformations, intra- and intermolecular interactions, and conformational dynamics of macromolecules. Especially for fluorescent-based single-molecule experiments, the choice of the fluorophore and labeling position are highly important. Here, we have studied the biophysical and structural effects that are associated with the conjugation of fluorophores to cysteines in the splicing factor U2AF65 by using single pair Förster resonance energy transfer (FRET) and nuclear magnetic resonance (NMR) spectroscopy...
August 10, 2017: Chemistry: a European Journal
https://www.readbyqxmd.com/read/28795702/cascade-photocaging-of-diazeniumdiolate-a-novel-strategy-for-one-and-two-photon-triggered-uncaging-with-real-time-reporting
#14
Krishna Kalyani Behara, Y Rajesh, Yarra Venkatesh, Bhaskar Rao Pinninti, Mahitosh Mandal, N D Pradeep Singh
We report a new strategy, viz. cascade photocaging, for protecting diethylamine diazeniumdiolate (O(2)-position), a light sensitive molecule. Upon photolysis, the cascade photocage at first releases the light activatable linker (latent fluorophore) O(2)-caged diazeniumdiolate, which undergoes spontaneous 1,8-elimination, triggering the release of the diazeniumdiolate anion and the fluorophore.
August 10, 2017: Chemical Communications: Chem Comm
https://www.readbyqxmd.com/read/28792742/sequential-release-of-proteins-from-structured-multishell-microcapsules
#15
Ulyana Shimanovich, Thomas C T Michaels, Erwin J de Genst, Dijana Matak-Vinkovic, Christopher M Dobson, Tuomas P J Knowles
In nature, a wide range of functional materials is based on proteins. Increasing attention is also turning to the use of proteins as artificial biomaterials in the form of films, gels, particles and fibrils that offer great potential for applications in areas ranging from molecular medicine to materials science. To date, however, most such applications have been limited to single component materials despite the fact that their natural analogues are composed of multiple types of proteins with a variety of functionalities that are co-assembled in a highly organized manner on the micron scale, a process which is currently challenging to achieve in the laboratory...
August 9, 2017: Biomacromolecules
https://www.readbyqxmd.com/read/28792728/an-environment-activatable-nanoprodrug-two-step-surveillance-in-the-anticancer-drug-release
#16
Sandipan Biswas, Joyjyoti Das, Shrabani Barman, Bhaskara Rao Pinninti, Tapas K Maiti, N D Pradeep Singh
Remedial cancer therapy deals with a large number of theranostic applications. However, systems, so far known, are only capable of single surveillance for both diagnostic and therapeutic modes of actions. A nanosystem, which can be localized to the cancer and deliver the chemotherapeutic agent on demand, will provide effective therapeutic activity. Herein, we designed a single component nanoprodrug ANPD-X (Activatable Nano Pro-Drug-X) which indentified the tumor sites by fluorescent colour change (signal 1, blue to green fluorescence) using H2O2-mediated oxidation of boronate fluorophore...
August 9, 2017: ACS Applied Materials & Interfaces
https://www.readbyqxmd.com/read/28791760/efficient-red-emissive-organic-crystals-with-amplified-spontaneous-emissions-based-on-a-single-benzene-framework
#17
Hongyu Zhang, Baolei Tang, Chenguang Wang, Yue Wang
Red-emissive fluorophores generally consist of large π-extended systems and thus encounter the problem of serious fluorescence quenching in solid state. Herein we reported a series of structurally simple compounds 2,5-bis(alkylamino)terephthalates 1a-c which consist of a very small π-system (only single benzene) but display efficient red emissions in crystals. The unique emission property of crystal 1a is mainly because of (1) the planarization of skeleton dominated by the strong intramolecular hydrogen bonds as well as (2) the unique 3-dimentional J-type molecular packing contributed by the mini π-system and the small dipole moment in excited state...
August 9, 2017: Angewandte Chemie
https://www.readbyqxmd.com/read/28789897/functionalization-of-peptide-nucleolipid-bioconjugates-and-their-structure-anti-cancer-activity-relationship-studies
#18
Niki Rana, Christopher Cultrara, Mariana Phillips, David Sabatino
In the search for more potent peptide-based anti-cancer conjugates the generation of new, functionally diverse nucleolipid derived D-(KLAKLAK)2-AK sequences has enabled a structure and anti-cancer activity relationship study. A reductive amination approach was key for the synthesis of alkylamine, diamine and polyamine derived nucleolipids as well as those incorporating heterocyclic functionality. The carboxy-derived nucleolipids were then coupled to the C-terminus of the D-(KLAKLAK)2-AK killer peptide sequence and produced with and without the FITC fluorophore for investigating biological activity in cancer cells...
July 26, 2017: Bioorganic & Medicinal Chemistry Letters
https://www.readbyqxmd.com/read/28789556/optical-determination-of-the-electronic-coupling-and-intercalation-geometry-of-thiazole-orange-homodimer-in-dna
#19
Paul D Cunningham, William P Bricker, Sebastián A Díaz, Igor L Medintz, Mark Bathe, Joseph S Melinger
Sequence-selective bis-intercalating dyes exhibit large increases in fluorescence in the presence of specific DNA sequences. This property makes this class of fluorophore of particular importance to biosensing and super-resolution imaging. Here we report ultrafast transient anisotropy measurements of resonance energy transfer (RET) between thiazole orange (TO) molecules in a complex formed between the homodimer TOTO and double-stranded (ds) DNA. Biexponential homo-RET dynamics suggest two subpopulations within the ensemble: 80% intercalated and 20% non-intercalated...
August 7, 2017: Journal of Chemical Physics
https://www.readbyqxmd.com/read/28783249/photophysics-electrochemistry-morphology-and-bioimaging-applications-of-new-1-8-naphthalimide-derivatives-containing-different-chromophores
#20
Ankita Saini, K R Justin Thomas, Abhay Sachdev, Packrisamy Gopinath
A series of 1,8-naphthalimide-based fluorophores containing different chromophores with varying conjugation and electron richness at imidic nitrogen are synthesized and characterized. These amine-functionalized naphthalimides are bipolar in nature and exhibit interesting optical and morphological variations attributable to the nature of N-substituents. Despite the dyes are structurally different due to the substituent variation in the imidic nitrogen, their electronic characteristics are similar originating from the 4-aminonaphthalimide segment...
August 7, 2017: Chemistry, An Asian Journal
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