Nile Red (nile + red)

Distribution by Scientific Domains

Terms modified by Nile Red

  • nile red staining

  • Selected Abstracts


    Adipogenic Effect of Alcohol on Human Bone Marrow-Derived Mesenchymal Stem Cells

    ALCOHOLISM, Issue 7 2004
    Frederick H. Wezeman
    Background: In addition to a decrease in bone mass in alcoholics their osteopenic skeletons show an increase in bone marrow adiposity. Human bone marrow mesenchymal stem cells (hMSC) in vivo differentiate into several phenotypes including osteogenic and adipogenic cells, both of which remain as resident populations of bone marrow. In vitro, the lineage commitment and differentiation of hMSC toward the adipogenic pathway can be promoted by alcohol. Methods: Human male and female mesenchymal stem cells from joint replacement surgery were cultured. Cells were grouped as: 1) Control (no additions to the culture medium), 2) EtOH (50 mm alcohol added to the culture medium), 3) OS (osteogenic inducers added to the culture medium), and 4) OS + EtOH (osteogenic inducers and 50 mm alcohol added to the culture medium). Cultures stained with Nile Red confirmed the development of differentiated adipocytes. Population analysis was performed using fluorescence-activated cell sorting. Gene expression of early, middle, late, and terminal differentiation stage markers (PPAR),2, lipoprotein lipase, adipsin, leptin, and adipocyte P2 (aP2)] was studied by Northern hybridization, and protein synthesis of aP2 was determined by Western analysis. Results: Nile red staining confirmed increased adipocyte development 10 days after the onset of treatment with 50 mm alcohol and osteogenic induction. By day 21 the number of adipocytes increased to 13.6% of the total population. Alcohol up-regulated the gene expression of PPAR,2 whereas no up-regulation was observed for the other genes. Protein production of aP2 was significantly increased in hMSC cells by culture in the presence of alcohol. Conclusions: The data suggest that alcohol's adipogenic effect on cultured hMSC is through up-regulation of PPAR,2 at the point of lineage commitment as well as through enhancement of lipid transport and storage through increased aP2 synthesis. The alcohol-induced expression and synthesis changes account for the increased Nile red staining of cultured hMSC. [source]


    An investigation of human brain tumour lipids by high-resolution magic angle spinning 1H MRS and histological analysis

    NMR IN BIOMEDICINE, Issue 7 2008
    Kirstie S. Opstad
    Abstract NMR-visible lipid signals detected in vivo by 1H MRS are associated with tumour aggression and believed to arise from cytoplasmic lipid droplets. High-resolution magic angle spinning (HRMAS) 1H MRS and Nile Red staining were performed on human brain tumour biopsy specimens to investigate how NMR-visible lipid signals relate to viable cells and levels of necrosis across different grades of glioma. Presaturation spectra were acquired from 24 adult human astrocytoma biopsy samples of grades II (8), III (2) and IV (14) using HRMAS 1H MRS and quantified using LCModel to determine lipid concentrations. Each biopsy sample was then refrozen, cryostat sectioned, and stained with Nile Red, to determine the number of lipid droplets and droplet size distribution, and with Haematoxylin and Eosin, to determine cell density and percentage necrosis. A strong correlation (R,=,0.92, P,<,0.0001) was found between the number of Nile Red-stained droplets and the ,1.3,ppm lipid proton concentration by 1H MRS. Droplet sizes ranged from 1 to 10,µm in diameter, and the size distribution was constant independent of tumour grade. In the non-necrotic biopsy samples, the number of lipid droplets correlated with cell density, whereas in the necrotic samples, there were greater numbers of droplets that showed a positive correlation with percentage necrosis. The correlation between 1H MRS lipid signals and number of Nile Red-stained droplets, and the presence of lipid droplets in the non-necrotic biopsy specimens provide good evidence that the in vivo NMR-visible lipid signals are cytoplasmic in origin and that formation of lipid droplets precedes necrosis. Copyright © 2008 John Wiley & Sons, Ltd. [source]


    Excited Singlet (S1)-state Interactions of Nile Red with Aromatic Amines,

    PHOTOCHEMISTRY & PHOTOBIOLOGY, Issue 2 2003
    J. Mohanty
    ABSTRACT Both steady-state (SS) and time-resolved (TR) studies show that the fluorescence of the dye Nile red (NR) is quenched by various aromatic amines (ArA). Bimolecular quenching constants (kq) from both SS and TR measurements are observed to match well, indicating that the interaction is dynamic in nature. The quenching interaction in the present systems has been attributed to electron transfer (ET) from ArA to excited NR, based on the variations in the kq values with the oxidation potentials of the amines. The kq values calculated within the framework of Marcus' outer-sphere ET theory at different free-energy changes (,G0) of the ET reactions match well with the experimental ones, supporting the ET mechanism in the systems studied. The reorganization energy (,) estimated from the correlation of the experimental and the calculated kq values is quite similar to the solvent reorganization energy (,s), calculated on the basis of the solvent dielectric continuum model along with the assumption that the reactants are the effective spheres. Although a modest error is involved in this ,s calculation, the similarity in , and ,s values suggests that the solvent reorganization plays a dominant role in governing the ET dynamics in the present systems. [source]


    Soil-borne wheat mosaic virus inclusion bodies: structural, compositional and staining properties

    ANNALS OF APPLIED BIOLOGY, Issue 2 2003
    L J LITTLEFIELD
    Summary Anatomy and cytochemistry of inclusion bodies induced by Soil-borne wheat mosaic virus infection were studied in roots and leaves to learn more about the nature of inclusions and their roles in pathogenesis. Acid Fuchsin, Giemsa stain, Toluidine Blue and Trypan Blue stains facilitated visualization of inclusion bodies. Combined, simultaneous staining with Acid Fuchsin and Toluidine Blue clearly differentiated inclusion bodies from host nuclei. The overall anatomy, composition and structure of virus inclusions in leaves and roots were generally similar, as shown by phase contrast, differential interference contrast, epifluorescence, laser scanning confocal and transmission electron microscopy. Both were often closely associated with host nuclei; both were comprised of intertwined masses of tubular material, presumably endoplasmic reticulum, and in which varied numbers and sizes of vacuolar cavities occurred. Leaf inclusions, however, were typically larger and more vacuolate than those in roots. Lipids were found to be significant constituents of both the tubular and vacuolar components of inclusions, indicated by positive staining with Nile Red and Sudan Black. Inclusion bodies in both leaves and roots lost their structural and compositional integrity, eventually becoming disorganized and devoid of clearly identifiable components as host tissue aged and symptom expression advanced. Significant results of this study include the first published examination of virus inclusion bodies in root tissue, the degree of structural detail of inclusion body anatomy revealed by laser scanning confocal microscopy and the presence of an extensive lipid component in virus inclusion bodies. [source]


    Activation of Sirt1 Decreases Adipocyte Formation During Osteoblast Differentiation of Mesenchymal Stem Cells,

    JOURNAL OF BONE AND MINERAL RESEARCH, Issue 7 2006
    Carl-Magnus Bäckesjö PhD
    Abstract In vitro, mesenchymal stem cells differentiate to osteoblasts when exposed to bone-inducing medium. However, adipocytes are also formed. We showed that activation of the nuclear protein deacetylase Sirt1 reduces adipocyte formation and promotes osteoblast differentiation. Introduction: Mesenchymal stem cells (MSCs) can differentiate into osteoblasts, adipocytes, chondrocytes, and myoblasts. It has been suggested that a reciprocal relationship exists between the differentiation of MSCs into osteoblasts and adipocytes. Peroxisome proliferator-activated receptor ,2 (PPAR,2) is a key element for the differentiation into adipocytes. Activation of Sirt1 has recently been shown to decrease adipocyte development from preadipocytes through inhibition of PPAR,2. Materials and Methods: We used the mouse mesenchymal cell line C3H10T1/2 and primary rat bone marrow cells cultured in osteoblast differentiation medium with or without reagents affecting Sirt1 activity. Adipocyte levels were analyzed by light microscopy and flow cytometry (FACS) after staining with Oil red O and Nile red, respectively. Osteoblast and adipocyte markers were studied with quantitative real-time PCR. Mineralization in cultures of primary rat bone marrow stromal cells was studied by von Kossa and alizarin red staining. Results: We found that Sirt1 is expressed in the mesenchymal cell line C3H10T1/2. Treatment with the plant polyphenol resveratrol as well as isonicotinamide, both of which activate Sirt1, blocked adipocyte development and increased the expression of osteoblast markers. Nicotinamide, which inhibits Sirt1, increased adipocyte number and increased expression of adipocyte markers. Furthermore, activation of Sirt1 prevented the increase in adipocytes caused by the PPAR,-agonist troglitazone. Finally, activation of Sirt1 in rat primary bone marrow stromal cells increased expression of osteoblast markers and also mineralization. Conclusions: In this study, we targeted Sirt1 to control adipocyte development during differentiation of MSCs into osteoblasts. The finding that resveratrol and isonicotinamide markedly inhibited adipocyte and promoted osteoblast differentiation may be relevant in the search for new treatment regimens of osteoporosis but also important for the evolving field of cell-based tissue engineering. [source]


    Excited Singlet (S1)-state Interactions of Nile Red with Aromatic Amines,

    PHOTOCHEMISTRY & PHOTOBIOLOGY, Issue 2 2003
    J. Mohanty
    ABSTRACT Both steady-state (SS) and time-resolved (TR) studies show that the fluorescence of the dye Nile red (NR) is quenched by various aromatic amines (ArA). Bimolecular quenching constants (kq) from both SS and TR measurements are observed to match well, indicating that the interaction is dynamic in nature. The quenching interaction in the present systems has been attributed to electron transfer (ET) from ArA to excited NR, based on the variations in the kq values with the oxidation potentials of the amines. The kq values calculated within the framework of Marcus' outer-sphere ET theory at different free-energy changes (,G0) of the ET reactions match well with the experimental ones, supporting the ET mechanism in the systems studied. The reorganization energy (,) estimated from the correlation of the experimental and the calculated kq values is quite similar to the solvent reorganization energy (,s), calculated on the basis of the solvent dielectric continuum model along with the assumption that the reactants are the effective spheres. Although a modest error is involved in this ,s calculation, the similarity in , and ,s values suggests that the solvent reorganization plays a dominant role in governing the ET dynamics in the present systems. [source]


    Investigation of binary polymer/surfactant or ternary polymer/surfactant/Cu2+ complexes in aqueous solution through Nile red probing

    POLYMER INTERNATIONAL, Issue 8 2010
    Zacharoula Iatridi
    Abstract The optical properties (absorption and emission) of Nile red have been widely used for staining or probing purposes in diverse aqueous systems. However, the applications of Nile red for the determination of the critical aggregation concentration of polymer/surfactant complexes or for the investigation of ternary polymer/surfactant/Cu2+ systems are very limited. The interactions between anionic polyelectrolytes and the oppositely charged surfactant N,N,N,N -dodecyltrimethylammonium chloride were investigated in dilute aqueous solution, exploiting the optical properties of Nile red. It is shown that the emission properties of Nile red present better sensitivity than its absorption properties, concerning the detection of the hydrophobic polymer/surfactant complexes formed in aqueous solution. Moreover, it is found that the formation of ternary polymer/surfactant/Cu2+ complexes leads to a pronounced quenching of the luminescence of Nile red. The corresponding Stern,Volmer plots indicate that quenching is more favourable when coordination of Cu2+ ions with poly(sodium acrylate) takes place, as compared to simple electrostatic binding of these ions with poly(sodium styrene sulfonate). Nile red is a sensitive and accurate tool, as an alternative to pyrene, for the characterization of binary polymer/surfactant complexes or for obtaining information on the local arrangement of ternary polymer/surfactant/Cu2+ systems in aqueous solution. Copyright © 2010 Society of Chemical Industry [source]


    A Convergent Approach to Biocompatible Polyglycerol "Click" Dendrons for the Synthesis of Modular Core,Shell Architectures and Their Transport Behavior

    CHEMISTRY - A EUROPEAN JOURNAL, Issue 30 2008
    Monika Wyszogrodzka Dipl.-Ing.
    Abstract Dendrimers are an important class of polymeric materials for a broad range of applications in which monodispersity and multivalency are of interest. Here we report on a highly efficient synthetic route towards bifunctional polyglycerol dendrons on a multigram scale. Commercially available triglycerol (1), which is highly biocompatible, was used as starting material. By applying Williamson ether synthesis followed by an ozonolysis/reduction procedure, glycerol-based dendrons up to the fourth generation were prepared. The obtained products have a reactive core, which was further functionalized to the corresponding monoazido derivatives. By applying copper(I)-catalyzed 1,3-dipolar cycloaddition, so-called "click" coupling, a library of core,shell architectures was prepared. After removal of the 1,2-diol protecting groups, water-soluble core,shell architectures 24,27 of different generations were obtained in high yields. In the structure,transport relationship with Nile red we observe a clear dependence on core size and generation of the polyglycerol dendrons. [source]


    Non-invasive Transdermal Delivery Route Using Electrostatically Interactive Biocompatible Nanocapsules

    ADVANCED MATERIALS, Issue 6 2010
    Jinseob Shin
    A robust means of fabricating skin-penetrating and compatible nanocarriers comprising poly(D, L -lactide- co -glycolide) is reported. The resultant nanocapsules have the ability to load biologically active ingredients and selectively release them through the epidermis lipid layer. Their release of genistein (left figure) is determined to be diffusion-controlled, and they are shown to successfully deliver molecules, such as nile red (right figure), into biopsied skin samples. [source]


    Skin permeability enhancement by low frequency sonophoresis: Lipid extraction and transport pathways

    JOURNAL OF PHARMACEUTICAL SCIENCES, Issue 6 2003
    R. Alvarez-Román
    Abstract The objective of this study was to shed light on the mechanism(s) by which low-frequency ultrasound (20 KHz) enhances the permeability of the skin. The physical effects on the barrier and the transport pathway, in particular, were examined. The amount of lipid removed from the intercellular domains of the stratum corneum following sonophoresis was determined by infrared spectroscopy. Transport of the fluorescent probes nile red and calcein, under the influence of ultrasound, was evaluated by laser-scanning confocal microscopy. The results were compared with the appropriate passive control data and with data obtained from experiments in which the skin was exposed simply to the thermal effects induced by ultrasound treatment. A significant fraction (,30%) of the intercellular lipids of the stratum corneum, which are principally responsible for skin barrier function, were removed during the application of low-frequency sonophoresis. Although the confocal images from the nile red experiments were not particularly informative, ultrasound clearly and significantly (again, relative to the corresponding controls) facilitated transport of the hydrophilic calcein via discrete permeabilized regions, whereas other areas of the barrier were apparently unaffected. Lipid removal from the stratum corneum is implicated as a factor contributing the observed permeation enhancement effects of low-frequency ultrasound. However, microscopic observations imply that sonophoresis induces localized (aqueous?) permeation pathways at discrete sites. © 2003 Wiley-Liss, Inc. and the American Pharmaceutical Association J Pharm Sci 92:1138,1146, 2003 [source]