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  1 / 7850 MEDLINE  
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PMID:29231937
Autor:Ugrinic M; Zambrano A; Berger S; Mann S; Tang TD; deMello A
Endereço:Department of Chemistry & Applied Biosciences, ETH Zurich, Vladimir Prelog Weg 1, 8093 Zurich, Switzerland. andrew.demello@chem.ethz.ch.
Título:Microfluidic formation of proteinosomes.
Fonte:Chem Commun (Camb); 54(3):287-290, 2018 Jan 02.
ISSN:1364-548X
País de publicação:England
Idioma:eng
Resumo:Herein we describe a novel microfluidic method for the generation of proteinosome micro-droplets, based on bovine serum albumin and glucose oxidase conjugated to PNIPAAm chains. The size of such water-in-oil droplets is regulated via control of the input reagent flow rate, with generated proteinosome populations exhibiting narrower size distributions than those observed when using standard bulk methodologies. Importantly, proteinosomes transferred from an oil to an aqueous-environment remain intact, become fully hydrated and exhibit an increase in average size. Moreover, functional proteinosomes prepared via microfluidics exhibit lower K values and higher enzymatic activities than proteinosomes produced by bulk methodologies.
Tipo de publicação: JOURNAL ARTICLE
Nome de substância:0 (Acrylic Resins); 25189-55-3 (poly-N-isopropylacrylamide); 27432CM55Q (Serum Albumin, Bovine); EC 1.1.3.4 (Glucose Oxidase); EC 1.11.1.- (Horseradish Peroxidase); I223NX31W9 (Fluorescein-5-isothiocyanate)


  2 / 7850 MEDLINE  
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PMID:29374708
Autor:Semkova S; Nikolova B; Zhelev Z; Tsoneva I; Zlateva G; Aoki I; Bakalova R
Endereço:Institute of Biophysics and Biomedical Engineering, Bulgarian Academy of Sciences, Sofia, Bulgaria.
Título:Loading Efficiency of Polymersomes with Contrast Agents and their Intracellular Delivery: Quantum Dots Organic Dyes.
Fonte:Anticancer Res; 38(2):825-831, 2018 02.
ISSN:1791-7530
País de publicação:Greece
Idioma:eng
Resumo:BACKGROUND/AIM: Contrast nanocarriers as drug-delivery systems, capable of selective delivery to cancer cells and solid tumors, are essential for the development of new diagnostic and therapeutic (theranostic) strategies. The present study aimed to investigate the loading efficiency of chitosan-based polymersomes with fluorescent contrast substances [quantum dots (QDs) and conventional organic dyes] and the possibility to control their release from the polymer matrix into cells by chemical modifications and electroporation. MATERIALS AND METHODS: All investigated fluorophores were retained within the polymer globule via electrostatic and hydrophilic-hydrophobic interactions, without conjugation with the polymer. The fluorophore-loaded polymersomes were characterized by dynamic light scattering, zeta-potential titration, and fluorescence spectroscopy. The release of fluorophore from the polymersomes, passively or after electroporation, was detected by 5-step spin-ultrafiltration, combined with fluorescence spectroscopy of the upper phase (supernatant) of the filter unit. Passive intracellular delivery of the nanoparticles to HeLa cells was detected by fluorescence confocal microscopy. RESULTS: The QDs were retained tightly and continuously in the polymer matrix, while the organic fluorophores [fluorescein isothiocyanate (FITC), FITC-dextran and FITC-dextran ] were released rapidly from the polymersomes. The detergent Brij significantly increased the retention of FITC-dextran in the polymer globule. Electroporation up to 1000 V/cm did not induce release of QDs from the polymersomes, but accelerated the release of Brij-treated FITC-dextran B from the polymer matrix. High-voltage pulses (over 750 V/cm) induced also fragmentation or aggregation of the nanoparticles. QD_labeled polymersomes penetrated passively in cancer cells after 24-hour incubation. CONCLUSION: The results suggest that QD-labeled polymersomes are appropriate fluorescent probes and a nano-drug delivery system with high tracing opportunities for in vitro and in vivo applications. Furthermore, loading polymersomes with organic dyes with different molecular weights (such as FITC-dextrans) is a simple model for visualizing and predicting the rate of release of small organic molecules (e.g. conventional drugs, other contrasts, stabilizers, and supplements) from the polymer matrix.
Tipo de publicação: COMPARATIVE STUDY; JOURNAL ARTICLE
Nome de substância:0 (Contrast Media); 0 (Dextrans); 0 (Fluorescent Dyes); 0 (fluorescein isothiocyanate dextran); 9012-76-4 (Chitosan); I223NX31W9 (Fluorescein-5-isothiocyanate)


  3 / 7850 MEDLINE  
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PMID:29198882
Autor:Sekiguchi K; Ogawa E; Kurohane K; Konishi H; Mochizuki N; Manabe K; Imai Y
Endereço:Laboratory of Microbiology and Immunology, School of Pharmaceutical Sciences, University of Shizuoka, 52-1 Yada, Suruga-ku, Shizuoka City, Shizuoka 422-8526, Japan.
Título:Adjuvant effect of short chain triacylglycerol tributyrin on a mouse contact hypersensitivity model.
Fonte:Toxicol Lett; 284:56-62, 2018 Mar 01.
ISSN:1879-3169
País de publicação:Netherlands
Idioma:eng
Resumo:Little attention has been paid to chemicals that can enhance hypersensitivity caused by other chemicals. We have demonstrated that phthalate esters with short chain alcohols enhance fluorescein isothiocyanate (FITC)-induced contact hypersensitivity (CHS) in a mouse model. Furthermore, phthalate esters with such an enhancing effect were found to activate transient receptor potential ankyrin 1 (TRPA1) cation channels, which are expressed on a part of sensory neurons, using a TRPA1-expressing cell line. In this study, we examined these activities of esters comprising glycerol and a short chain fatty acid, i.e. dibutyrin and tributyrin. We carried out chemical synthesis of dibutyrin isomers. Each dibutyrin isomer weakly activated TRPA1 and slightly enhanced skin sensitization to FITC. Unexpectedly, TRPA1 activation and enhancement of FITC-CHS were much more evident in the presence of tributyrin. Mechanistically, tributyrin induced increased dendritic cell trafficking from the skin to draining lymph nodes. Tributyrin enhanced interferon-γ (IFN-γ) production by draining lymph nodes, while its effect on interleukin-4 (IL-4) production was relatively less prominent. These results suggested that tributyrin concomitantly caused TRPA1 activation and an adjuvant effect on FITC-CHS.
Tipo de publicação: JOURNAL ARTICLE
Nome de substância:0 (Adjuvants, Immunologic); 0 (TRPA1 Cation Channel); 0 (Triglycerides); 207137-56-2 (Interleukin-4); 82115-62-6 (Interferon-gamma); I223NX31W9 (Fluorescein-5-isothiocyanate); S05LZ624MF (tributyrin)


  4 / 7850 MEDLINE  
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PMID:28456983
Autor:Eriksson I; Öllinger K; Appelqvist H
Endereço:Experimental Pathology, Department of Clinical and Experimental Medicine, Linköping University, SE-58185, Linköping, Sweden.
Título:Analysis of Lysosomal pH by Flow Cytometry Using FITC-Dextran Loaded Cells.
Fonte:Methods Mol Biol; 1594:179-189, 2017.
ISSN:1940-6029
País de publicação:United States
Idioma:eng
Resumo:The acidic environment of the lysosomal lumen provides an optimal milieu for the acid hydrolases and is also essential for fusion/fission of endo-lysosomal compartments and sorting of cargo. Evidence suggests that maintaining lysosomal acidity is essential to avoid disease. In this chapter, we describe a protocol for analyzing the lysosomal pH in cultured cells using the fluorescent probe fluorescein isothiocyanate (FITC)-dextran together with a dual-emission ratiometric technique suitable for flow cytometry. Fluorescence-labeled dextran is endocytosed and accumulated in the lysosomal compartment. FITC shows a pH-dependent variation in fluorescence when analyzed at maximum emission wavelength and no variation when analyzing at the isosbestic point, thereby the ratio can be used to determine the lysosomal pH. A standard curve is obtained by equilibrating intralysosomal pH with extracellular pH using the ionophore nigericin. The protocol also includes information regarding procedures to induce lysosomal alkalinization and lysosomal membrane permeabilization.
Tipo de publicação: JOURNAL ARTICLE
Nome de substância:0 (Dextrans); 0 (Macrolides); 0 (fluorescein isothiocyanate dextran); 116764-51-3 (bafilomycin A); I223NX31W9 (Fluorescein-5-isothiocyanate)


  5 / 7850 MEDLINE  
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PMID:28973323
Autor:Mathieu E; Gupta N; Ahari A; Zhou X; Hanna J; Yücel YH
Endereço:Keenan Research Centre for Biomedical Science, Li Ka Shing Knowledge Institute, St. Michael's Hospital, Toronto, Ontario, Canada.
Título:Evidence for Cerebrospinal Fluid Entry Into the Optic Nerve via a Glymphatic Pathway.
Fonte:Invest Ophthalmol Vis Sci; 58(11):4784-4791, 2017 Sep 01.
ISSN:1552-5783
País de publicação:United States
Idioma:eng
Resumo:Purpose: The purpose of this study was to determine whether cerebrospinal fluid (CSF) enters the optic nerve via a glymphatic pathway and whether this entry is size-dependent. Methods: Fluorescent dextran tracers (fluorescein isothiocyanate [FITC]) of four different sizes (10, 40, 70, and 500 kDa) and FITC-ovalbumin (45 kDa) were injected into the CSF of 15 adult mice. Tracer distribution in the orbital optic nerve at 1 hour after injection was assessed in tissue sections with confocal microscopy. Tracer distribution within the optic nerve was studied in relation to blood vessels and astrocytes identified by isolectin histochemistry and glial fibrillary acidic protein (GFAP) immunofluorescence, respectively. Aquaporin 4 (AQP4) immunostaining was performed to assess astrocytic endfeet in relation to CSF tracer. Results: One hour following tracer injection into CSF, all tracer sizes (10-500 kDa) were noted in the subarachnoid space surrounding the orbital optic nerve. In all cases, 10 kDa (n = 4/4) and 40 kDa (n = 3/3) tracers were noted within the optic nerve, while 70-kDa tracer was occasionally noted (n = 1/4). Tracer found within the nerve was specifically localized between isolectin-labeled blood vessels and GFAP-positive astrocytes or AQP4-labeled astrocytic endfeet. The 500-kDa tracer was not detected within the optic nerve. Conclusions: To our knowledge, this is the first evidence of a glymphatic pathway in the optic nerve. CSF enters the optic nerve via spaces surrounding blood vessels, bordered by astrocytic endfeet. CSF entry into paravascular spaces of the optic nerve is size-dependent, and this pathway may be highly relevant to optic nerve diseases, including glaucoma.
Tipo de publicação: JOURNAL ARTICLE
Nome de substância:0 (Dextrans); 0 (Fluorescent Dyes); 0 (fluorescein isothiocyanate dextran); I223NX31W9 (Fluorescein-5-isothiocyanate)


  6 / 7850 MEDLINE  
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PMID:28935759
Autor:Karlsen TV; Reikvam T; Tofteberg A; Nikpey E; Skogstrand T; Wagner M; Tenstad O; Wiig H
Endereço:From the Department of Biomedicine, University of Bergen, Norway (T.V.K., T.R., A.T., E.N., T.S., M.W., O.T., H.W.); and Departments of Medicine (E.N.) and Pathology (M.W.), Haukeland University Hospital, Bergen, Norway. tine.karlsen@uib.no helge.wiig@uib.no.
Título:Lymphangiogenesis Facilitates Initial Lymph Formation and Enhances the Dendritic Cell Mobilizing Chemokine CCL21 Without Affecting Migration.
Fonte:Arterioscler Thromb Vasc Biol; 37(11):2128-2135, 2017 Nov.
ISSN:1524-4636
País de publicação:United States
Idioma:eng
Resumo:OBJECTIVE: Lymphatic vessels play an important role in body fluid, as well as immune system homeostasis. Although the role of malfunctioning or missing lymphatics has been studied extensively, less is known on the functional consequences of a chronically expanded lymphatic network or lymphangiogenesis. APPROACH AND RESULTS: To this end, we used K14-VEGF-C (keratin-14 vascular endothelial growth factor-C) transgenic mice overexpressing the vascular endothelial growth factor C in skin and investigated the responses to inflammatory and fluid volume challenges. We also recorded interstitial fluid pressure, a major determinant of lymph flow. Transgenic mice had a strongly enhanced lymph vessel area in skin. Acute inflammation induced by lipopolysaccharide and chronic inflammation by delayed-type hypersensitivity both resulted in increased interstitial fluid pressure and reduced lymph flow, both to the same extent in wild-type and transgenic mice. Hyperplastic lymphatic vessels, however, demonstrated enhanced transport capacity after local fluid overload not induced by inflammation. In this situation, interstitial fluid pressure was increased to a similar extent in the 2 strains, thus, suggesting that the enhanced lymph vessel area facilitated initial lymph formation. The increased lymph vessel area resulted in an enhanced production of the chemoattractant CCL21 that, however, did not result in augmented dendritic cell migration after induction of local skin inflammation by fluorescein isothiocyanate. CONCLUSIONS: An expanded lymphatic network is capable of enhanced chemoattractant production, and lymphangiogenesis will facilitate initial lymph formation favoring increased clearance of fluid in situations of augmented fluid filtration.
Tipo de publicação: JOURNAL ARTICLE
Nome de substância:0 (Chemokine CCL21); 0 (Keratin-14); 0 (Lipopolysaccharides); 0 (Vascular Endothelial Growth Factor C); 15646-46-5 (Oxazolone); I223NX31W9 (Fluorescein-5-isothiocyanate)


  7 / 7850 MEDLINE  
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PMID:28800593
Autor:Mussawy H; Viezens L; Hauenherm G; Schroeder M; Schaefer C
Endereço:Department of Orthopaedic Surgery, University Medical Centre Hamburg-Eppendorf, Hamburg, Germany.
Título:In vivo functional and morphological characterization of bone and striated muscle microcirculation in NSG mice.
Fonte:PLoS One; 12(8):e0183186, 2017.
ISSN:1932-6203
País de publicação:United States
Idioma:eng
Resumo:Organ-specific microcirculation plays a central role in tumor growth, tumor cell homing, tissue engineering, and wound healing. Mouse models are widely used to study these processes; however, these mouse strains often possess unique microhemodynamic parameters, making it difficult to directly compare experiments. The full functional characterization of bone and striated muscle microcirculatory parameters in non-obese diabetic-severe combined immunodeficiency/y-chain; NOD-Prkds IL2rg (NSG) mice has not yet been reported. Here, we established either a dorsal skinfold chamber or femur window in NSG mice (n = 23), allowing direct analysis of microcirculatory parameters in vivo by intravital fluorescence microscopy at 7, 14, 21, and 28 days after chamber preparation. Organ-specific differences were observed. Bone had a significantly lower vessel density but a higher vessel diameter than striated muscle. Bone also showed higher effective vascular permeability than striated muscle. The centerline velocity values were similar in the femur window and dorsal skinfold chamber, with a higher volumetric blood flow in bone. Interestingly, bone and striated muscle showed similar tissue perfusion rates. Knowledge of physiological microhemodynamic values of bone and striated muscle in NSG mice makes it possible to analyze pathophysiological processes at these anatomic sites, such as tumor growth, tumor metastasis, and tumor microcirculation, as well as the response to therapeutic agents.
Tipo de publicação: JOURNAL ARTICLE
Nome de substância:0 (Fluorescent Dyes); 0 (fluorescein isothiocyanate bovine serum albumin); 27432CM55Q (Serum Albumin, Bovine); I223NX31W9 (Fluorescein-5-isothiocyanate)


  8 / 7850 MEDLINE  
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PMID:28768154
Autor:Sun YQ; Dai CM; Zheng Y; Shi SD; Hu HY; Chen DW
Endereço:School of Pharmacy, Jinzhou Medical University, Jinzhou, PR China; School of Pharmacy, Shenyang Pharmaceutical University, Shenyang, PR China.
Título:Binding effect of fluorescence labeled glycyrrhetinic acid with GA receptors in hepatocellular carcinoma cells.
Fonte:Life Sci; 188:186-191, 2017 Nov 01.
ISSN:1879-0631
País de publicação:Netherlands
Idioma:eng
Resumo:Glycyrrhetinic acid (GA) is a natural active component from licorice, which is broadly used in traditional Chinese medicine. Lots of glycyrrhetinic acid receptors (GA-R) are proved to locate on the surface of liver cells. Many reports about the hepatocellular carcinoma (HCC) treatment were dependent on GA modified carriers. However, the reality of GA-R in HCC cells was not clear. In this paper, 18ß-glycyrrhetinic acid (18ß-GA) was labeled with fluorescence (FITC) by chemical synthesis. Together with the binding effect of fluorescence labeled glycyrrhetinic acid (FITC-GA), the competitive action of 18ß-GA with GA-R was investigated in HCC cells. The results showed that in HepG2 cells, 18ß-GA and FITC-GA presented similar cytotoxicity. The specific binding saturation of GA showed the dissociation constant (K ) was 7.457±2.122pmol/L and the maximum binding counts (B ) was 2.385±0.175pmol/2.5×10 cells, respectively. FITC-GA bound to cytomembrane specifically and 18ß-GA competed to bind the sites significantly in HepG2 cells. Therefore, there is binding effect between fluorescence labeled GA and GA-R. The GA-R on HCC cells is confirmed as expected, which provides a useful reference of active target modified by GA and a novel approach for receptors and ligands study.
Tipo de publicação: JOURNAL ARTICLE
Nome de substância:0 (Ligands); 1449-05-4 (18alpha-glycyrrhetinic acid); I223NX31W9 (Fluorescein-5-isothiocyanate); P540XA09DR (Glycyrrhetinic Acid)


  9 / 7850 MEDLINE  
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PMID:28759570
Autor:Rezaee F; Harford TJ; Linfield DT; Altawallbeh G; Midura RJ; Ivanov AI; Piedimonte G
Endereço:Pediatric Research Center and Pediatric Institute, Cleveland Clinic Children's, Cleveland, Ohio, United States of America.
Título:cAMP-dependent activation of protein kinase A attenuates respiratory syncytial virus-induced human airway epithelial barrier disruption.
Fonte:PLoS One; 12(7):e0181876, 2017.
ISSN:1932-6203
País de publicação:United States
Idioma:eng
Resumo:Airway epithelium forms a barrier to the outside world and has a crucial role in susceptibility to viral infections. Cyclic adenosine monophosphate (cAMP) is an important second messenger acting via two intracellular signaling molecules: protein kinase A (PKA) and the guanidine nucleotide exchange factor, Epac. We sought to investigate effects of increased cAMP level on the disruption of model airway epithelial barrier caused by RSV infection and the molecular mechanisms underlying cAMP actions. Human bronchial epithelial cells were infected with RSV-A2 and treated with either cAMP releasing agent, forskolin, or cAMP analogs. Structure and functions of the Apical Junctional Complex (AJC) were evaluated by measuring transepithelial electrical resistance and permeability to FITC-dextran, and determining localization of AJC proteins by confocal microscopy. Increased intracellular cAMP level significantly attenuated RSV-induced disassembly of AJC. These barrier-protective effects of cAMP were due to the activation of PKA signaling and did not involve Epac activity. Increased cAMP level reduced RSV-induced reorganization of the actin cytoskeleton, including apical accumulation of an essential actin-binding protein, cortactin, and inhibited expression of the RSV F protein. These barrier-protective and antiviral-function of cAMP signaling were evident even when cAMP level was increased after the onset of RSV infection. Taken together, our study demonstrates that cAMP/PKA signaling attenuated RSV-induced disruption of structure and functions of the model airway epithelial barrier by mechanisms involving the stabilization of epithelial junctions and inhibition of viral biogenesis. Improving our understanding of the mechanisms involved in RSV-induced epithelial dysfunction and viral pathogenesis will help to develop novel anti-viral therapeutic approaches.
Tipo de publicação: JOURNAL ARTICLE
Nome de substância:0 (Dextrans); 0 (Guanine Nucleotide Exchange Factors); 0 (Microfilament Proteins); 0 (fluorescein isothiocyanate dextran); 1F7A44V6OU (Colforsin); E0399OZS9N (Cyclic AMP); EC 2.7.11.11 (Cyclic AMP-Dependent Protein Kinases); I223NX31W9 (Fluorescein-5-isothiocyanate)


  10 / 7850 MEDLINE  
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PMID:28737767
Autor:Fernandez-Carrera A; Vigo E; Regueiro-Rodríguez C; González-Fernández Á; Olivieri D; Aroeira LS
Endereço:Immunology, Biomedical Research Center (CINBIO) (Centro Singular de Investigación de Galicia 2016-2019) and Galicia-Sur Health Research Institute (IIS-GS), University Campus, University of Vigo, Vigo, Spain.
Título:Sensitive and non-invasive method for the in vivo analysis of membrane permeability in small animals.
Fonte:Lab Invest; 97(9):1114-1120, 2017 Sep.
ISSN:1530-0307
País de publicação:United States
Idioma:eng
Resumo:Tissue membranes are boundaries that isolate organs or cavities in the body. These semi-permeable membranes are responsible for passive protection that acts through the regulation of nutrient absorption, secretion and filtration of small molecules. These functions could be altered as a consequence of inflammation or trauma, which in turn could lead to changes in permeability, allowing the entrance of toxins, antigens, proteins or facilitating the spread of tumors. Membrane permeability therefore plays an important role in numerous diseases. However, current experimental techniques that are available to quantify membrane permeability in small animals have limited precision and temporal specificity. Improvements in such measurements would lead to a deeper understanding of disease pathogenesis and this may accelerate the development of specific therapies. The study reported here concerns the efficacy of a novel, non-invasive imaging analysis-based measurement method that significantly improves the quantification of tissue membrane permeability in small animals, while at the same time mitigating the adverse effects experienced by the animals under study.
Tipo de publicação: JOURNAL ARTICLE
Nome de substância:0 (Dextrans); 0 (Dialysis Solutions); 0 (fluorescein isothiocyanate dextran); I223NX31W9 (Fluorescein-5-isothiocyanate)



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