Base de dados : MEDLINE
Pesquisa : D02.092.782.420 [Categoria DeCS]
Referências encontradas : 327 [refinar]
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[PMID]:26634306
[Au] Autor:Song Y; Li J; Li D
[Ad] Endereço:Department of Marine Engineering, Dalian Maritime University, Dalian, P. R. China.
[Ti] Título:Zeta potentials of polydimethylsiloxane surfaces modified by polybrene of different concentrations.
[So] Source:Electrophoresis;37(4):567-72, 2016 Feb.
[Is] ISSN:1522-2683
[Cp] País de publicação:Germany
[La] Idioma:eng
[Ab] Resumo:Zeta potential is an important parameter for characterizing the electrokinetic properties of a solid-liquid interface. In this paper, zeta potentials of polydimethylsiloxane surfaces modified by polybrene (PB) solutions of different concentrations in Phosphate buffer solution and pure water were reported. The zeta potentials were measured by an induction current method. The measurements were validated both by a calibration curve based on the data reported in the published papers and by comparing the zeta potential determined by using the Smoluchowski equation and the measured velocity of the electrokinetic motion of particles in a microchannel.
[Mh] Termos MeSH primário: Dimetilpolisiloxanos/química
Condutividade Elétrica
Técnicas Eletroquímicas/instrumentação
Brometo de Hexadimetrina/química
[Mh] Termos MeSH secundário: Desenho de Equipamento
Propriedades de Superfície
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (Dimethylpolysiloxanes); 4C905MSK4W (Hexadimethrine Bromide); 63148-62-9 (baysilon)
[Em] Mês de entrada:1612
[Cu] Atualização por classe:161230
[Lr] Data última revisão:
161230
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:151205
[St] Status:MEDLINE
[do] DOI:10.1002/elps.201500465


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[PMID]:26555613
[Au] Autor:Pajarinen J; Lin TH; Sato T; Loi F; Yao Z; Konttinen YT; Goodman SB
[Ad] Endereço:Orthopaedic Research Laboratories, Department of Orthopaedic Surgery, Stanford University School of Medicine, Stanford, CA, United States of America.
[Ti] Título:Establishment of Green Fluorescent Protein and Firefly Luciferase Expressing Mouse Primary Macrophages for In Vivo Bioluminescence Imaging.
[So] Source:PLoS One;10(11):e0142736, 2015.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Macrophages play a key role in tissue homeostasis as well as in a range of pathological conditions including atherosclerosis, cancer, and autoimmunity. Many aspects of their in vivo behavior are, however, poorly understood. Bioluminescence imaging (BLI) with green fluorescent protein (GFP) and firefly luciferase (FLUC) labelled autologous reporter macrophages could potentially offer a powerful tool to study macrophage biology, but this approach has been hindered by the relative difficulty of efficient gene transfer into primary macrophages. Here we describe a straightforward method for producing large numbers of GFP/FLUC expressing mouse primary macrophages utilizing lentivirus vector, cyclosporine, and a double infection strategy. Using this method we achieved up to 60% of macrophages to express GFP with correspondingly high FLUC signal. When injected into the circulation using a mouse model of local biomaterial induced inflammation and osteolysis, macrophages were initially detectable within the lungs, followed by systemic homing to the local area of chronic inflammation in the distal femur. In addition, transduced macrophages maintained their ability to assume M1 and M2 phenotypes although the GFP/FLUC expression was altered by the polarizing signals. These reporter macrophages could prove to be valuable tools to study the role of macrophages in health and disease.
[Mh] Termos MeSH primário: Proteínas de Fluorescência Verde/genética
Luciferases de Vaga-Lume/genética
Macrófagos/metabolismo
Imagem Molecular
[Mh] Termos MeSH secundário: Animais
Células Cultivadas
Ciclosporina/farmacologia
Dextranos/farmacologia
Brometo de Hexadimetrina/farmacologia
Luminescência
Macrófagos/citologia
Macrófagos/efeitos dos fármacos
Masculino
Camundongos
Camundongos Endogâmicos BALB C
Transdução Genética
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, N.I.H., EXTRAMURAL; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (Dextrans); 147336-22-9 (Green Fluorescent Proteins); 4C905MSK4W (Hexadimethrine Bromide); 83HN0GTJ6D (Cyclosporine); EC 1.13.12.7 (Luciferases, Firefly)
[Em] Mês de entrada:1606
[Cu] Atualização por classe:170220
[Lr] Data última revisão:
170220
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:151112
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0142736


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[PMID]:26071903
[Au] Autor:Han M; Yu D; Song Q; Wang J; Dong P; He J
[Ad] Endereço:Department of Otorhinolaryngology, Shanghai First People's Hospital, Shanghai Jiao Tong University, Shanghai 200080, China.
[Ti] Título:Polybrene: Observations on cochlear hair cell necrosis and minimal lentiviral transduction of cochlear hair cells.
[So] Source:Neurosci Lett;600:164-70, 2015 Jul 23.
[Is] ISSN:1872-7972
[Cp] País de publicação:Ireland
[La] Idioma:eng
[Ab] Resumo:Polybrene is widely used to enhance viral transduction; however, little is known about the utility thereof, in enhancing lentiviral transduction of cochlear cells. In the present study, we examined the cytotoxic effects of polybrene, and the further effects thereof, on lentiviral transduction of cochlear cells, especially sensory hair cells. Cochlear basilar membranes of newborn rats were cultured and treated with 0.1-10 µg/mL polybrene for 24h to explore the potential development of ototoxicity. PI staining and TUNEL detection were used to evaluate necrosis or apoptosis of hair cell. Various doses of lentivirus-GFP were added to cochlear organotypic cultures with safe concentrations of polybrene, incubated for 24h, and cultured (in the absence of the virus and polybrene) for a further 48 h. Transduction efficiencies were evaluated. The results showed that polybrene at 0.1 µg/mL was safe to cochlear cells, and 0.5-10 µg/mL concentration induced hair cell necrosis in a dose-dependent manner. However, supporting cells were not damaged. Lentiviral vectors transduced into cochlear cells and 0.1 µg/mL polybrene enhanced transduction efficiency. However, hair cells were hardly transduced with lentiviral vectors either alone or in the presence of 0.1 µg/mL polybrene. The use of polybrene to aid lentiviral transduction of cochlear hair cells requires further attention.
[Mh] Termos MeSH primário: Células Ciliadas Auditivas/efeitos dos fármacos
Brometo de Hexadimetrina/toxicidade
Lentivirus/genética
Transdução Genética
[Mh] Termos MeSH secundário: Animais
Animais Recém-Nascidos
Apoptose
Vetores Genéticos
Proteínas de Fluorescência Verde/genética
Proteínas de Fluorescência Verde/metabolismo
Células Ciliadas Auditivas/metabolismo
Células Ciliadas Auditivas/patologia
Necrose
Ratos Sprague-Dawley
Técnicas de Cultura de Tecidos
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (enhanced green fluorescent protein); 147336-22-9 (Green Fluorescent Proteins); 4C905MSK4W (Hexadimethrine Bromide)
[Em] Mês de entrada:1510
[Cu] Atualização por classe:150718
[Lr] Data última revisão:
150718
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:150615
[St] Status:MEDLINE


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[PMID]:25643596
[Au] Autor:Zhang Z; Miao Y; Zhang Q; Lian L; Yan G
[Ad] Endereço:Shanxi Normal University, Linfen, Shanxi 041000, China.
[Ti] Título:Selective room temperature phosphorescence detection of heparin based on manganese-doped zinc sulfide quantum dots/polybrene self-assembled nanosensor.
[So] Source:Biosens Bioelectron;68:556-62, 2015 Jun 15.
[Is] ISSN:1873-4235
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:A selective system was developed to detect heparin in aqueous solutions by using MPA(3-Mercaptopropionic Acid)-capped Mn-doped ZnS quantum dots (QDs)/polybrene (hexadimethrine bromide) hybrids as a sensitive room temperature phosphorescence (RTP) nanosensor. In this system, the RTP intensity of QDs was remarkably enhanced via electrostatic self-assembly after the addition of polybrene. The addition of heparin into the system was competitively bound to polybrene and enable to deprive it from the surface of QDs, as a result, the RTP intensity of Mn-doped ZnS QDs/polybrene hybrids was reduced with the increased of heparin concentration. Based on this effect, a selective system was proposed to detect heparin. Under the optimal conditions, the change of RTP intensity was proportional to the heparin concentration from 0.05 to 1.4 U mL(-1) (about 0.38-10.76 µg mL(-1)) and the limit of detection (LOD) was 0.021 U mL(-1) (about 0.16 µg mL(-1)). This proposed nanosensor is simple and relatively free of interference from coexisting substances, which can be applied to detect heparin in heparin injection and human serum. In addition, a new pathway was also provided based on the assembly of QDs with other cationic homopolymers for further design of biosensors and detection of biomolecules.
[Mh] Termos MeSH primário: Técnicas Biossensoriais
Heparina/isolamento & purificação
Medições Luminescentes
[Mh] Termos MeSH secundário: Heparina/química
Brometo de Hexadimetrina/química
Seres Humanos
Manganês/química
Nanoestruturas/química
Pontos Quânticos
Sulfetos/química
Temperatura Ambiente
Compostos de Zinco/química
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (Sulfides); 0 (Zinc Compounds); 42Z2K6ZL8P (Manganese); 4C905MSK4W (Hexadimethrine Bromide); 9005-49-6 (Heparin); KPS085631O (zinc sulfide)
[Em] Mês de entrada:1511
[Cu] Atualização por classe:150302
[Lr] Data última revisão:
150302
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:150204
[St] Status:MEDLINE


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[PMID]:25260343
[Au] Autor:Pei L; Lucy CA
[Ad] Endereço:Department of Chemistry, Gunning/Lemieux Chemistry Centre, University of Alberta, Edmonton, AB, Canada T6G 2G2.
[Ti] Título:Insight into the stability of poly(diallydimethylammoniumchloride) and polybrene poly cationic coatings in capillary electrophoresis.
[So] Source:J Chromatogr A;1365:226-33, 2014 Oct 24.
[Is] ISSN:1873-3778
[Cp] País de publicação:Netherlands
[La] Idioma:eng
[Ab] Resumo:Polycationic polymers are widely used in capillary electrophoresis (CE) as surface coatings to prevent protein adsorption and control electroosmotic flow (EOF). Such semi-permanent coatings are formed by flushing the capillary with a quaternary amine-based polymer such as poly(diallydimethylammonium chloride) (PDADMAC) or polybrene. Compared to covalent capillary coatings, the claimed advantages of adsorptive polycation coatings are their simple preparation and that they are not limited to the pH 2-8 range as are covalent coatings. However, while the latter is commonly claimed, few studies have demonstrated the stability of polycationic coatings at extreme pH. Herein PDADMAC and polybrene are studied as model cationic coatings. PDADMAC with higher molecular weight (M.W.) demonstrated higher EOF stability at pH 9.5, with PDADMAC of M.W. less than 200,000 being unstable at pH 9.5. X-ray photoelectron spectroscopy (XPS) shows that the quaternary amines of PDADMAC and polybrene were slowly converted to tertiary amines in alkaline solution and more rapidly when adsorbed on a silica surface. The degraded polycation deprotonated at pH >7, resulting in loss of polymer from the surface and diminishing EOF. Successive multiple ionic layer (SMIL) coatings show greater alkaline stability by distancing the polycation from the surface. Separations of inorganic anions at pH 9.5 illustrate the degradation behavior and enhanced stability of higher M.W. polycationic coatings.
[Mh] Termos MeSH primário: Eletroforese Capilar/instrumentação
Polietilenos/química
Compostos de Amônio Quaternário/química
[Mh] Termos MeSH secundário: Adsorção
Ânions
Cátions
Eletro-Osmose
Eletroforese Capilar/métodos
Brometo de Hexadimetrina
Concentração de Íons de Hidrogênio
Espectroscopia Fotoeletrônica
Dióxido de Silício/química
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (Anions); 0 (Cations); 0 (Polyethylenes); 0 (Quaternary Ammonium Compounds); 26062-79-3 (poly-N,N-dimethyl-N,N-diallylammonium chloride); 4C905MSK4W (Hexadimethrine Bromide); 7631-86-9 (Silicon Dioxide)
[Em] Mês de entrada:1412
[Cu] Atualização por classe:141006
[Lr] Data última revisão:
141006
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:140928
[St] Status:MEDLINE


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[PMID]:24801942
[Au] Autor:Brandl C; Zimmermann SJ; Milenkovic VM; Rosendahl SM; Grassmann F; Milenkovic A; Hehr U; Federlin M; Wetzel CH; Helbig H; Weber BH
[Ad] Endereço:Institute of Human Genetics, University of Regensburg, Franz-Josef-Strauss-Allee 11, 93053, Regensburg, Germany.
[Ti] Título:In-depth characterisation of Retinal Pigment Epithelium (RPE) cells derived from human induced pluripotent stem cells (hiPSC).
[So] Source:Neuromolecular Med;16(3):551-64, 2014 Sep.
[Is] ISSN:1559-1174
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Induced pluripotent stem cell (iPSC)-derived retinal pigment epithelium (RPE) has widely been appreciated as a promising tool to model human ocular disease emanating from primary RPE pathology. Here, we describe the successful reprogramming of adult human dermal fibroblasts to iPSCs and their differentiation to pure expandable RPE cells with structural and functional features characteristic for native RPE. Fibroblast cultures were established from skin biopsy material and subsequently reprogrammed following polycistronic lentiviral transduction with OCT4, SOX2, KLF4 and L-Myc. Fibroblast-derived iPSCs showed typical morphology, chromosomal integrity and a distinctive stem cell marker profile. Subsequent differentiation resulted in expandable pigmented hexagonal RPE cells. The cells revealed stable RNA expression of mature RPE markers RPE65, RLBP and BEST1. Immunolabelling verified localisation of BEST1 at the basolateral plasma membrane, and scanning electron microscopy showed typical microvilli at the apical side of iPSC-derived RPE cells. Transepithelial resistance was maintained at high levels during cell culture indicating functional formation of tight junctions. Secretion capacity was demonstrated for VEGF-A. Feeding of porcine photoreceptor outer segments revealed the proper ability of these cells for phagocytosis. IPSC-derived RPE cells largely maintained these properties after cryopreservation. Together, our study underlines that adult dermal fibroblasts can serve as a valuable resource for iPSC-derived RPE with characteristics highly reminiscent of true RPE cells. This will allow its broad application to establish cellular models for RPE-related human diseases.
[Mh] Termos MeSH primário: Células Epiteliais/citologia
Células-Tronco Pluripotentes Induzidas/citologia
Epitélio Pigmentado da Retina/citologia
[Mh] Termos MeSH secundário: Adulto
Animais
Antígenos de Diferenciação/análise
Diferenciação Celular
Separação Celular
Células Cultivadas
Técnicas de Cocultura
Criopreservação
Meios de Cultura/farmacologia
Células Epiteliais/metabolismo
Células Epiteliais/secreção
Proteínas do Olho/biossíntese
Proteínas do Olho/genética
Feminino
Fibroblastos/citologia
Fibroblastos/efeitos dos fármacos
Brometo de Hexadimetrina/farmacologia
Seres Humanos
Células-Tronco Pluripotentes Induzidas/efeitos dos fármacos
Cariotipagem
Lentivirus/fisiologia
Camundongos
Microvilosidades/ultraestrutura
Fagocitose
RNA Mensageiro/biossíntese
RNA Mensageiro/genética
Segmento Externo da Célula Bastonete
Pele/citologia
Sus scrofa
Preservação de Tecido
Fator A de Crescimento do Endotélio Vascular/secreção
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (Antigens, Differentiation); 0 (Culture Media); 0 (Eye Proteins); 0 (RNA, Messenger); 0 (VEGFA protein, human); 0 (Vascular Endothelial Growth Factor A); 4C905MSK4W (Hexadimethrine Bromide)
[Em] Mês de entrada:1510
[Cu] Atualização por classe:140804
[Lr] Data última revisão:
140804
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:140508
[St] Status:MEDLINE
[do] DOI:10.1007/s12017-014-8308-8


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[PMID]:24658746
[Au] Autor:Zhao C; Wu N; Deng F; Zhang H; Wang N; Zhang W; Chen X; Wen S; Zhang J; Yin L; Liao Z; Zhang Z; Zhang Q; Yan Z; Liu W; Wu D; Ye J; Deng Y; Zhou G; Luu HH; Haydon RC; Si W; He TC
[Ad] Endereço:Departments of Clinical Hematology, Cell Biology and Oncology, the Affiliated Southwest Hospital of the Third Military Medical University, Chongqing, China; Molecular Oncology Laboratory, Department of Orthopaedic Surgery, The University of Chicago Medical Center, Chicago, Illinois, United States of
[Ti] Título:Adenovirus-mediated gene transfer in mesenchymal stem cells can be significantly enhanced by the cationic polymer polybrene.
[So] Source:PLoS One;9(3):e92908, 2014.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Mesenchymal stem cells (MSCs) are multipotent progenitors, which can undergo self-renewal and give rise to multi-lineages. A great deal of attentions have been paid to their potential use in regenerative medicine as potential therapeutic genes can be introduced into MSCs. Genetic manipulations in MSCs requires effective gene deliveries. Recombinant adenoviruses are widely used gene transfer vectors. We have found that although MSCs can be infected in vitro by adenoviruses, high virus titers are needed to achieve high efficiency. Here, we investigate if the commonly-used cationic polymer Polybrene can potentiate adenovirus-mediated transgene delivery into MSCs, such as C2C12 cells and iMEFs. Using the AdRFP adenovirus, we find that AdRFP transduction efficiency is significantly increased by Polybrene in a dose-dependent fashion peaking at 8 µg/ml in C2C12 and iMEFs cells. Quantitative luciferase assay reveals that Polybrene significantly enhances AdFLuc-mediated luciferase activity in C2C12 and iMEFs at as low as 4 µg/ml and 2 µg/ml, respectively. FACS analysis indicates that Polybrene (at 4 µg/ml) increases the percentage of RFP-positive cells by approximately 430 folds in AdRFP-transduced iMEFs, suggesting Polybrene may increase adenovirus infection efficiency. Furthermore, Polybrene can enhance AdBMP9-induced osteogenic differentiation of MSCs as early osteogenic marker alkaline phosphatase activity can be increased more than 73 folds by Polybrene (4 µg/ml) in AdBMP9-transduced iMEFs. No cytotoxicity was observed in C2C12 and iMEFs at Polybrene up to 40 µg/ml, which is about 10-fold higher than the effective concentration required to enhance adenovirus transduction in MSCs. Taken together, our results demonstrate that Polybrene should be routinely used as a safe, effective and inexpensive augmenting agent for adenovirus-mediated gene transfer in MSCs, as well as other types of mammalian cells.
[Mh] Termos MeSH primário: Adenoviridae/genética
Técnicas de Transferência de Genes
Vetores Genéticos/genética
Brometo de Hexadimetrina/farmacologia
Células Mesenquimais Estromais/efeitos dos fármacos
Células Mesenquimais Estromais/metabolismo
Transdução Genética
[Mh] Termos MeSH secundário: Animais
Linhagem Celular
Relação Dose-Resposta a Droga
Citometria de Fluxo
Expressão Gênica
Genes Reporter
Seres Humanos
Camundongos
Transgenes
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, N.I.H., EXTRAMURAL; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
4C905MSK4W (Hexadimethrine Bromide)
[Em] Mês de entrada:1411
[Cu] Atualização por classe:170220
[Lr] Data última revisão:
170220
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:140325
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0092908


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[PMID]:24626396
[Au] Autor:Haselberg R; Oliveira S; van der Meel R; Somsen GW; de Jong GJ
[Ad] Endereço:Biomolecular Analysis, Utrecht University, Universiteitsweg 99, 3584 CG Utrecht, The Netherlands; Division of BioAnalytical Chemistry, AIMMS research group BioMolecular Analysis, VU University Amsterdam, De Boelelaan 1083, 1081 HV Amsterdam, The Netherlands. Electronic address: r.haselberg@vu.nl.
[Ti] Título:Capillary electrophoresis-based assessment of nanobody affinity and purity.
[So] Source:Anal Chim Acta;818:1-6, 2014 Mar 25.
[Is] ISSN:1873-4324
[Cp] País de publicação:Netherlands
[La] Idioma:eng
[Ab] Resumo:Drug purity and affinity are essential attributes during development and production of therapeutic proteins. In this work, capillary electrophoresis (CE) was used to determine both the affinity and composition of the biotechnologically produced "nanobody" EGa1, the binding fragment of a heavy-chain-only antibody. EGa1 is an antagonist of the epidermal growth factor receptor (EGFR), which is overexpressed on the surface of tumor cells. Using a background electrolyte (BGE) of 50mM sodium phosphate (pH 8.0) in combination with a polybrene-poly(vinylsulfonic acid) capillary coating, CE analysis of EGa1 showed the presence of at least three components. Affinity of the EGa1 components towards the extracellular domain of EGFR was assessed by adding different concentrations (0-12 nM) of the receptor to the BGE while measuring the effective electrophoretic mobility of the respective EGa1 components. Binding curves obtained by plotting electrophoretic mobility shifts as a function of receptor concentration, yielded dissociation constants (Kd) of 1.65, 1.67, and 1.75 nM for the three components, respectively; these values were comparable to the Kd of 2.1 nM obtained for the bulk EGa1 product using a cellular assay. CE with mass spectrometry (MS) detection using a BGE of 25 mM ammonium acetate (pH 8.0) revealed that the EGa1 sample comprised of significant amounts of deamidated, bisdeamidated and N-terminal pyroglutamic acid products. CE-MS using a BGE of 100mM acetic acid (pH 2.8) in combination with a polybrene-dextran sulfate-polybrene capillary coating demonstrated the additional presence of minor products related to incomplete removal of the signal peptide from the produced nanobody. Combining the results obtained from affinity CE and CE-MS, it is concluded that the EGa1 nanobody product is heterogeneous, comprising highly-related proteins that exhibit very similar affinity towards EGFR.
[Mh] Termos MeSH primário: Técnicas de Química Analítica/métodos
Eletroforese Capilar
Anticorpos de Domínio Único/análise
[Mh] Termos MeSH secundário: Acetatos/química
Ácido Acético/química
Sequência de Aminoácidos
Sulfato de Dextrana/química
Brometo de Hexadimetrina/química
Cinética
Espectrometria de Massas
Dados de Sequência Molecular
Fosfatos/química
Polivinil/química
Receptor do Fator de Crescimento Epidérmico/antagonistas & inibidores
Receptor do Fator de Crescimento Epidérmico/imunologia
Anticorpos de Domínio Único/imunologia
Anticorpos de Domínio Único/metabolismo
Ácidos Sulfônicos/química
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (Acetates); 0 (Phosphates); 0 (Polyvinyls); 0 (Single-Domain Antibodies); 0 (Sulfonic Acids); 26101-52-0 (lyapolate); 4C905MSK4W (Hexadimethrine Bromide); 9042-14-2 (Dextran Sulfate); EC 2.7.10.1 (Receptor, Epidermal Growth Factor); Q40Q9N063P (Acetic Acid); RRE756S6Q2 (ammonium acetate); SE337SVY37 (sodium phosphate)
[Em] Mês de entrada:1411
[Cu] Atualização por classe:140314
[Lr] Data última revisão:
140314
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:140315
[St] Status:MEDLINE


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[PMID]:24281865
[Au] Autor:Ramirez VP; Aneskievich BJ
[Ad] Endereço:Graduate Program in Pharmacology & Toxicology, Department of Pharmaceutical Sciences, School of Pharmacy, University of Connecticut, Storrs, CT, USA.
[Ti] Título:Transgene delivery to cultured keratinocytes via replication-deficient adenovirus vectors.
[So] Source:Methods Mol Biol;1195:43-8, 2014.
[Is] ISSN:1940-6029
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Transient transgene expression can facilitate investigation of that gene-product function or effect on keratinocyte biology. Several chemical and biologic delivery systems are available, and among them adenoviruses offer particular advantages in efficiency and transgene capacity. Here we describe the advantages of bicistronic adenovirus and inclusion of the polycation hexadimethrine bromide to aid in the detection of positively transduced cells and enhance transduction efficiency.
[Mh] Termos MeSH primário: Adenoviridae/genética
Vetores Genéticos/genética
Queratinócitos/citologia
Queratinócitos/metabolismo
Transdução Genética/métodos
Transgenes/genética
[Mh] Termos MeSH secundário: Linhagem Celular
Análise Custo-Benefício
Meios de Cultura
Genes Reporter/genética
Brometo de Hexadimetrina/metabolismo
Seres Humanos
Transdução Genética/economia
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, N.I.H., EXTRAMURAL; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (Culture Media); 4C905MSK4W (Hexadimethrine Bromide)
[Em] Mês de entrada:1502
[Cu] Atualização por classe:140712
[Lr] Data última revisão:
140712
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:131128
[St] Status:MEDLINE
[do] DOI:10.1007/7651_2013_43


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[PMID]:24406242
[Au] Autor:Nanba D; Matsushita N; Toki F; Higashiyama S
[Ti] Título:Efficient expansion of human keratinocyte stem/progenitor cells carrying a transgene with lentiviral vector.
[So] Source:Stem Cell Res Ther;4(5):127, 2013 Oct 18.
[Is] ISSN:1757-6512
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:INTRODUCTION: The development of an appropriate procedure for lentiviral gene transduction into keratinocyte stem cells is crucial for stem cell biology and regenerative medicine for genetic disorders of the skin. However, there is little information available on the efficiency of lentiviral transduction into human keratinocyte stem/progenitor cells and the effects of gene transduction procedures on growth potential of the stem cells by systematic assessment. METHODS: In this study, we explored the conditions for efficient expansion of human keratinocyte stem/progenitor cells carrying a transgene with a lentiviral vector, by using the culture of keratinocytes on a feeder layer of 3 T3 mouse fibroblasts. The gene transduction and expansion of keratinocytes carrying a transgene were analyzed by Western blotting, quantitative PCR, and flow cytometry. RESULTS: Polybrene (hexadiamine bromide) markedly enhanced the efficiency of lentiviral gene transduction, but negatively affected the maintenance of the keratinocyte stem/progenitor cells at a concentration higher than 5 µg/ml. Rho-assiciated kinase (ROCK) inhibitor Y-27632, a small molecule which enhanced keratinocyte proliferation, significantly interfered with the lentiviral transduction into cultured human keratinocytes. However, a suitable combination of polybrene and Y-27632 effectively expanded keratinocytes carrying a transgene. CONCLUSIONS: This study provides information for effective expansion of cultured human keratinocyte stem/progenitor cells carrying a transgene. This point is particularly significant for the application of genetically modified keratinocyte stem/progenitor stem cells in regenerative medicine.
[Mh] Termos MeSH primário: Vetores Genéticos/metabolismo
Queratinócitos/citologia
Lentivirus/genética
Células-Tronco/citologia
Transgenes/genética
[Mh] Termos MeSH secundário: Células 3T3
Amidas/farmacologia
Animais
Diferenciação Celular/efeitos dos fármacos
Proliferação Celular/efeitos dos fármacos
Inibidores Enzimáticos/farmacologia
Vetores Genéticos/genética
Proteínas de Fluorescência Verde/genética
Proteínas de Fluorescência Verde/metabolismo
Antagonistas de Heparina/farmacologia
Brometo de Hexadimetrina/farmacologia
Seres Humanos
Camundongos
Piridinas/farmacologia
Células-Tronco/metabolismo
Transfecção
Quinases Associadas a rho/antagonistas & inibidores
Quinases Associadas a rho/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (Amides); 0 (Enzyme Inhibitors); 0 (Heparin Antagonists); 0 (Pyridines); 0 (enhanced green fluorescent protein); 138381-45-0 (Y 27632); 147336-22-9 (Green Fluorescent Proteins); 4C905MSK4W (Hexadimethrine Bromide); EC 2.7.11.1 (rho-Associated Kinases)
[Em] Mês de entrada:1409
[Cu] Atualização por classe:150515
[Lr] Data última revisão:
150515
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:140111
[St] Status:MEDLINE
[do] DOI:10.1186/scrt338



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