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Pesquisa : D02.455.326.271.665.325 [Categoria DeCS]
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[PMID]:29317633
[Au] Autor:Guo P; Liu D; Subramanyam K; Wang B; Yang J; Huang J; Auguste DT; Moses MA
[Ad] Endereço:Vascular Biology Program, Boston Children's Hospital, 300 Longwood Avenue, Boston, MA, 02115, USA.
[Ti] Título:Nanoparticle elasticity directs tumor uptake.
[So] Source:Nat Commun;9(1):130, 2018 01 09.
[Is] ISSN:2041-1723
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:To date, the role of elasticity in drug delivery remains elusive due to the inability to measure microscale mechanics and alter rheology without affecting chemistry. Herein, we describe the in vitro cellular uptake and in vivo tumor uptake of nanolipogels (NLGs). NLGs are composed of identical lipid bilayers encapsulating an alginate core, with tunable elasticity. The elasticity of NLGs was evaluated by atomic force microscopy, which demonstrated that they exhibit Young's moduli ranging from 45 ± 9 to 19,000 ± 5 kPa. Neoplastic and non-neoplastic cells exhibited significantly greater uptake of soft NLGs (Young's modulus <1.6 MPa) relative to their elastic counterparts (Young's modulus >13.8 MPa). In an orthotopic breast tumor model, soft NLGs accumulated significantly more in tumors, whereas elastic NLGs preferentially accumulated in the liver. Our findings demonstrate that particle elasticity directs tumor accumulation, suggesting that it may be a design parameter to enhance tumor delivery efficiency.
[Mh] Termos MeSH primário: Neoplasias da Mama/metabolismo
Módulo de Elasticidade
Nanopartículas/química
Nanopartículas/metabolismo
[Mh] Termos MeSH secundário: Animais
Neoplasias da Mama/patologia
Linhagem Celular
Linhagem Celular Tumoral
Clorpromazina/farmacologia
Endocitose/efeitos dos fármacos
Filipina/farmacologia
Seres Humanos
Hidrazonas/farmacologia
Fígado/metabolismo
Células MCF-7
Neoplasias Mamárias Experimentais/metabolismo
Camundongos Endogâmicos BALB C
Microscopia de Força Atômica
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, N.I.H., EXTRAMURAL; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (Hydrazones); 0 (N'-(3,4-dihydroxybenzylidene)-3-hydroxy-2-naphthahydrazide); 87Z59R7D14 (Filipin); U42B7VYA4P (Chlorpromazine)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180226
[Lr] Data última revisão:
180226
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:180111
[St] Status:MEDLINE
[do] DOI:10.1038/s41467-017-02588-9


  2 / 729 MEDLINE  
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[PMID]:28813441
[Au] Autor:Gehne N; Lamik A; Lehmann M; Haseloff RF; Andjelkovic AV; Blasig IE
[Ad] Endereço:Leibniz-Forschungsinstitut für Molekulare Pharmakologie, Berlin, Germany.
[Ti] Título:Cross-over endocytosis of claudins is mediated by interactions via their extracellular loops.
[So] Source:PLoS One;12(8):e0182106, 2017.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Claudins (Cldns) are transmembrane tight junction (TJ) proteins that paracellularly seal endo- and epithelial barriers by their interactions within the TJs. However, the mechanisms allowing TJ remodeling while maintaining barrier integrity are largely unknown. Cldns and occludin are heterophilically and homophilically cross-over endocytosed into neighboring cells in large, double membrane vesicles. Super-resolution microscopy confirmed the presence of Cldns in these vesicles and revealed a distinct separation of Cldns derived from opposing cells within cross-over endocytosed vesicles. Colocalization of cross-over endocytosed Cldn with the autophagosome markers as well as inhibition of autophagosome biogenesis verified involvement of the autophagosomal pathway. Accordingly, cross-over endocytosed Cldns underwent lysosomal degradation as indicated by lysosome markers. Cross-over endocytosis of Cldn5 depended on clathrin and caveolin pathways but not on dynamin. Cross-over endocytosis also depended on Cldn-Cldn-interactions. Amino acid substitutions in the second extracellular loop of Cldn5 (F147A, Q156E) caused impaired cis- and trans-interaction, as well as diminished cross-over endocytosis. Moreover, F147A exhibited an increased mobility in the membrane, while Q156E was not as mobile but enhanced the paracellular permeability. In conclusion, the endocytosis of TJ proteins depends on their ability to interact strongly with each other in cis and trans, and the mobility of Cldns in the membrane is not necessarily an indicator of barrier permeability. TJ-remodeling via cross-over endocytosis represents a general mechanism for the degradation of transmembrane proteins in cell-cell contacts and directly links junctional membrane turnover to autophagy.
[Mh] Termos MeSH primário: Clatrina/metabolismo
Claudinas/metabolismo
Endocitose/fisiologia
[Mh] Termos MeSH secundário: Animais
Caveolina 1/metabolismo
Linhagem Celular
Clorpromazina/farmacologia
Claudina-3/metabolismo
Claudinas/química
Claudinas/genética
Cães
Endocitose/efeitos dos fármacos
Endocitose/genética
Filipina/farmacologia
Seres Humanos
Imuno-Histoquímica
Camundongos
Ocludina/metabolismo
Ligação Proteica/genética
Ligação Proteica/fisiologia
Transdução de Sinais/efeitos dos fármacos
Junções Íntimas/efeitos dos fármacos
Junções Íntimas/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Caveolin 1); 0 (Clathrin); 0 (Claudin-3); 0 (Claudins); 0 (Occludin); 87Z59R7D14 (Filipin); U42B7VYA4P (Chlorpromazine)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171013
[Lr] Data última revisão:
171013
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170817
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0182106


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[PMID]:28630013
[Au] Autor:Kunz D; Oliveira GB; Uchôa AF; Samuels RI; Macedo MLR; Silva CP
[Ad] Endereço:Departamento de Bioquímica, Centro de Ciências Biológicas, Universidade Federal de Santa Catarina, C.P. 476, 88040-900 Florianópolis, SC, Brazil.
[Ti] Título:Receptor mediated endocytosis of vicilin in Callosobruchus maculatus (Coleoptera: Chrysomelidae) larval midgut epithelial cells.
[So] Source:Comp Biochem Physiol B Biochem Mol Biol;210:39-47, 2017 Aug.
[Is] ISSN:1879-1107
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:The transport of proteins across the intestinal epithelium of insects is still not well understood. There is evidence that vicilin, a major storage protein of cowpea seeds (Vigna unguiculata), is internalized in larvae of the seed-beetle Callosobruchus maculatus. It has been reported that this vicilin interacts with proteins present in the microvillar membranes of columnar cells along the digestive tract of the larvae. In the present work, we studied the cellular pathway involved in endocytosis of vicilin in larval C. maculatus by employing ex vivo experiments. In the ex vivo approach, we incubated FITC-labelled vicilin with isolated midgut wholemounts in the absence or in the presence of endocytosis inhibitors. The fate of labelled or non-labelled globulins was monitored by confocal microscopy and fluorescence measurement. Our results suggest that the internalization of vicilins is due to receptor-mediated endocytosis. Here we report the identity of a microvillar vicilin-binding protein that was purified using affinity chromatography on a vicilin-sepharose column. The putative vicilin receptor showed high homology to proteins with the CRAL-TRIO domain, specifically the Sec14 superfamily member α-tocopherol transfer protein. The precise mechanism involved in vicilin internalization was defined through the use of specific inhibitors of the endocytosis pathway. The inhibitors filipin III and nystatin significantly inhibited the endocytosis of vicilin, while chlorpromazine and phenylarsine oxide had a much lower effect on endocytosis, suggesting that the endocytic pathway is predominantly mediated by caveolin.
[Mh] Termos MeSH primário: Proteínas de Transporte/metabolismo
Coleópteros/metabolismo
Células Epiteliais/metabolismo
Proteínas de Insetos/metabolismo
Larva/metabolismo
Proteínas de Armazenamento de Sementes/metabolismo
[Mh] Termos MeSH secundário: Sequência de Aminoácidos
Animais
Arsenicais/farmacologia
Transporte Biológico
Proteínas de Transporte/genética
Clorpromazina/farmacologia
Coleópteros/efeitos dos fármacos
Coleópteros/genética
Sistema Digestório/efeitos dos fármacos
Sistema Digestório/metabolismo
Sistema Digestório/ultraestrutura
Endocitose/efeitos dos fármacos
Endocitose/genética
Células Epiteliais/efeitos dos fármacos
Células Epiteliais/ultraestrutura
Filipina/farmacologia
Fluoresceína-5-Isotiocianato/química
Corantes Fluorescentes/química
Expressão Gênica
Proteínas de Insetos/genética
Larva/efeitos dos fármacos
Larva/genética
Nistatina/farmacologia
Proteínas de Armazenamento de Sementes/genética
Alinhamento de Sequência
Homologia de Sequência de Aminoácidos
Coloração e Rotulagem
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Arsenicals); 0 (Carrier Proteins); 0 (Fluorescent Dyes); 0 (Insect Proteins); 0 (Seed Storage Proteins); 0 (alpha-tocopherol transfer protein); 0HUR2WY345 (oxophenylarsine); 1400-61-9 (Nystatin); 87Z59R7D14 (Filipin); 9067-60-1 (vicilin protein, plant); I223NX31W9 (Fluorescein-5-isothiocyanate); U42B7VYA4P (Chlorpromazine)
[Em] Mês de entrada:1707
[Cu] Atualização por classe:171116
[Lr] Data última revisão:
171116
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170621
[St] Status:MEDLINE


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[PMID]:27710850
[Au] Autor:Belli V; Guarnieri D; Biondi M; Della Sala F; Netti PA
[Ad] Endereço:Center for Advanced Biomaterial for Health Care (CABHC), Istituto Italiano di Tecnologia, Largo Barsanti e Matteucci 53, Napoli, Italy.
[Ti] Título:Dynamics of nanoparticle diffusion and uptake in three-dimensional cell cultures.
[So] Source:Colloids Surf B Biointerfaces;149:7-15, 2017 Jan 01.
[Is] ISSN:1873-4367
[Cp] País de publicação:Netherlands
[La] Idioma:eng
[Ab] Resumo:This study aims at elucidating the effect of three-dimensional (3D) extracellular matrix on cell behaviour and nanoparticle (NP) diffusion and its consequences on NP cellular uptake mechansims. For this purpose, human dermal fibroblasts (HDF) and human fibrosarcoma (HT1080) cell lines were grown within a 3D collagen gel and exposed to model polystyrene (PS) NPs of controlled size (44 and 100nm). Results indicate that, in 3D, cell morphology dramatically changes compared to standard 2D cultures and NP diffusion within the matrix is hampered by the interaction with the collagen fibres. As a consequence, NP cellular uptake, modeled with equations describing the stoichiometric exchange between NPs and cell membrane, is significantly slowed down in 3D and in the case of 100 nm NPs, in part due to the hampered diffusion of NPs in collagen gel compared to their transport in standard cell culture medium. Furthermore, our outcomes point at a significant contribution of the cytoskeleton assembly, in particular actin microfilaments, in governing the uptake of PS NPs in a 3D environment, and also that the macropinocytosis process is preserved and is mainly involved in the internalization of PS NPs in a 3D environment. However, depending on cell type and nanoparticle size, other endocytic pathways are also implicated when moving from 2D to 3D culture systems. This work highlights the importance of studying the nano-bio interaction in experimental models that resembles in vivo conditions in order to better predict the therapeutic efficacy of drug delivery systems.
[Mh] Termos MeSH primário: Citoesqueleto de Actina/efeitos dos fármacos
Fibroblastos/efeitos dos fármacos
Nanopartículas/metabolismo
Poliestirenos/farmacologia
[Mh] Termos MeSH secundário: Citoesqueleto de Actina/metabolismo
Citoesqueleto de Actina/ultraestrutura
Técnicas de Cultura de Células
Linhagem Celular
Linhagem Celular Tumoral
Membrana Celular/efeitos dos fármacos
Membrana Celular/metabolismo
Membrana Celular/ultraestrutura
Sobrevivência Celular/efeitos dos fármacos
Colágeno/química
Difusão
Endocitose/efeitos dos fármacos
Fibroblastos/metabolismo
Fibroblastos/ultraestrutura
Filipina/farmacologia
Géis
Seres Humanos
Cinética
Nanopartículas/química
Nocodazol/farmacologia
Tamanho da Partícula
Poliestirenos/química
Sacarose/farmacologia
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Gels); 0 (Polystyrenes); 57-50-1 (Sucrose); 87Z59R7D14 (Filipin); 9007-34-5 (Collagen); SH1WY3R615 (Nocodazole)
[Em] Mês de entrada:1702
[Cu] Atualização por classe:170224
[Lr] Data última revisão:
170224
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:161007
[St] Status:MEDLINE


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[PMID]:27234403
[Au] Autor:Ribas GS; Souza HM; de Mari J; Deon M; Mescka C; Saraiva-Pereira ML; Kessler R; Trapp F; Michelin K; Burin M; Vargas CR; Giugliani R
[Ad] Endereço:Programa de Pós-graduação em Saúde da Criança e do Adolescente, Universidade Federal do Rio Grande do Sul (UFRGS), Porto Alegre, RS, Brazil; Serviço de Genética Médica, Hospital de Clínicas de Porto Alegre, Porto Alegre, RS, Brazil. Electronic address: grazielaribas@yahoo.com.br.
[Ti] Título:Selective screening of Niemann-Pick type C Brazilian patients by cholestane-3ß,5α,6ß-triol and chitotriosidase measurements followed by filipin staining and NPC1/NPC2 gene analysis.
[So] Source:Clin Chim Acta;459:57-62, 2016 Aug 01.
[Is] ISSN:1873-3492
[Cp] País de publicação:Netherlands
[La] Idioma:eng
[Mh] Termos MeSH primário: Proteínas de Transporte/genética
Colestanóis/sangue
Filipina/química
Glicoproteínas/genética
Hexosaminidases/sangue
Glicoproteínas de Membrana/genética
Doença de Niemann-Pick Tipo C/diagnóstico
Coloração e Rotulagem
[Mh] Termos MeSH secundário: 1-Desoxinojirimicina/análogos & derivados
1-Desoxinojirimicina/uso terapêutico
Brasil
Hexosaminidases/metabolismo
Seres Humanos
Doença de Niemann-Pick Tipo C/sangue
Doença de Niemann-Pick Tipo C/tratamento farmacológico
Doença de Niemann-Pick Tipo C/genética
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Carrier Proteins); 0 (Cholestanols); 0 (Glycoproteins); 0 (Membrane Glycoproteins); 0 (NPC1 protein, human); 0 (NPC2 protein, human); 115510-05-9 (cholestane-3,5,6-triol); 19130-96-2 (1-Deoxynojirimycin); 87Z59R7D14 (Filipin); ADN3S497AZ (miglustat); EC 3.2.1.- (Hexosaminidases); EC 3.2.1.- (chitotriosidase)
[Em] Mês de entrada:1702
[Cu] Atualização por classe:170826
[Lr] Data última revisão:
170826
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:160529
[St] Status:MEDLINE


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[PMID]:27181747
[Au] Autor:Hamatani M; Jingami N; Uemura K; Nakasone N; Kinoshita H; Yamakado H; Ninomiya H; Takahashi R
[Ad] Endereço:Department of Neurology, Kyoto University Graduate School of Medicine.
[Ti] Título:A case of variant biochemical phenotype of Niemann-Pick disease type C accompanying savant syndrome.
[So] Source:Rinsho Shinkeigaku;56(6):424-9, 2016 06 22.
[Is] ISSN:1882-0654
[Cp] País de publicação:Japan
[La] Idioma:jpn
[Ab] Resumo:A 40-year-old man was referred to our hospital because of vertical supranuclear gaze palsy, frequent sudden loss of muscle tonus and ataxia for several years. He had a history of prolonged neonatal jaundice. He was given a diagnosis of autism in his childhood, followed by a diagnosis of schizophrenia in his teenage. He also developed a savant skill of calendar calculating. (123)I-IMP-SPECT showed decreased cerebral blood flow in the left frontotemporal lobe as often seen in savant syndrome. Although genetic analysis of NPC1 and NPC2 revealed no pathogenic mutation, filipin staining of cultured fibroblasts from his biopsied skin revealed a certain amount of intracellular cholesterol storage pattern, indicating a variant biochemical phenotype of Niemann-Pick disease type C (NPC). The diagnosis of adulthood onset NPC is difficult and challenging, especially for neurologists, because the symptoms and signs are not as clear as those in the classical childhood onset NPC and this subtype is not yet widely known. However, the diagnosis can be made by a combination of filipin staining of fibroblast and/or gene analysis. As a disease-specific therapy for NPC has been approved in Japan, the diagnosis of NPC is of significance.
[Mh] Termos MeSH primário: Transtorno Autístico/complicações
Transtorno Autístico/diagnóstico
Doença de Niemann-Pick Tipo C/complicações
Doença de Niemann-Pick Tipo C/diagnóstico
Fenótipo
Esquizofrenia/complicações
Esquizofrenia/diagnóstico
[Mh] Termos MeSH secundário: Adulto
Transtorno Autístico/genética
Transtorno Autístico/fisiopatologia
Fibroblastos
Filipina
Testes Genéticos
Seres Humanos
Masculino
Doença de Niemann-Pick Tipo C/genética
Doença de Niemann-Pick Tipo C/fisiopatologia
Esquizofrenia/genética
Esquizofrenia/fisiopatologia
Coloração e Rotulagem
Síndrome
Tomografia Computadorizada de Emissão de Fóton Único
[Pt] Tipo de publicação:CASE REPORTS; JOURNAL ARTICLE
[Nm] Nome de substância:
87Z59R7D14 (Filipin)
[Em] Mês de entrada:1703
[Cu] Atualização por classe:171012
[Lr] Data última revisão:
171012
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:160517
[St] Status:MEDLINE
[do] DOI:10.5692/clinicalneurol.cn-000846


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[PMID]:27033412
[Au] Autor:Rezgui R; Blumer K; Yeoh-Tan G; Trexler AJ; Magzoub M
[Ad] Endereço:Biology Program, New York University Abu Dhabi, Abu Dhabi, United Arab Emirates.
[Ti] Título:Precise quantification of cellular uptake of cell-penetrating peptides using fluorescence-activated cell sorting and fluorescence correlation spectroscopy.
[So] Source:Biochim Biophys Acta;1858(7 Pt A):1499-506, 2016 Jul.
[Is] ISSN:0006-3002
[Cp] País de publicação:Netherlands
[La] Idioma:eng
[Ab] Resumo:Cell-penetrating peptides (CPPs) have emerged as a potentially powerful tool for drug delivery due to their ability to efficiently transport a whole host of biologically active cargoes into cells. Although concerted efforts have shed some light on the cellular internalization pathways of CPPs, quantification of CPP uptake has proved problematic. Here we describe an experimental approach that combines two powerful biophysical techniques, fluorescence-activated cell sorting (FACS) and fluorescence correlation spectroscopy (FCS), to directly, accurately and precisely measure the cellular uptake of fluorescently-labeled molecules. This rapid and technically simple approach is highly versatile and can readily be applied to characterize all major CPP properties that normally require multiple assays, including amount taken up by cells (in moles/cell), uptake efficiency, internalization pathways, intracellular distribution, intracellular degradation and toxicity threshold. The FACS-FCS approach provides a means for quantifying any intracellular biochemical entity, whether expressed in the cell or introduced exogenously and transported across the plasma membrane.
[Mh] Termos MeSH primário: Membrana Celular/metabolismo
Peptídeos Penetradores de Células/análise
Coloração e Rotulagem/métodos
[Mh] Termos MeSH secundário: Cloreto de Amônio/farmacologia
Biotina/química
Membrana Celular/efeitos dos fármacos
Permeabilidade da Membrana Celular/efeitos dos fármacos
Peptídeos Penetradores de Células/metabolismo
Clorpromazina/farmacologia
Citocalasina D/farmacologia
Endocitose/efeitos dos fármacos
Filipina/farmacologia
Citometria de Fluxo
Corantes Fluorescentes/química
Células HeLa
Seres Humanos
Cinética
Transporte Proteico/efeitos dos fármacos
Espectrometria de Fluorescência/métodos
Estreptavidina/química
Succinimidas/química
beta-Ciclodextrinas/farmacologia
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (Alexa Fluor 488 carboxylic acid succinimidyl ester); 0 (Cell-Penetrating Peptides); 0 (Fluorescent Dyes); 0 (Succinimides); 0 (beta-Cyclodextrins); 0 (methyl-beta-cyclodextrin); 01Q9PC255D (Ammonium Chloride); 22144-77-0 (Cytochalasin D); 6SO6U10H04 (Biotin); 87Z59R7D14 (Filipin); 9013-20-1 (Streptavidin); U42B7VYA4P (Chlorpromazine)
[Em] Mês de entrada:1608
[Cu] Atualização por classe:161126
[Lr] Data última revisão:
161126
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:160402
[St] Status:MEDLINE


  8 / 729 MEDLINE  
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[PMID]:26878859
[Au] Autor:Wang H; Liu W; Yu F; Lu L
[Ad] Endereço:National Pathogen Collection Center for Aquatic Animals, Shanghai Ocean university, Shanghai, 201306, China. haowang1000@163.com.
[Ti] Título:Disruption of clathrin-dependent trafficking results in the failure of grass carp reovirus cellular entry.
[So] Source:Virol J;13:25, 2016 Feb 16.
[Is] ISSN:1743-422X
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:BACKGROUND: Grass carp reovirus (GCRV) is responsible for viral hemorrhagic disease in cultured grass carp (Ctenopharyngon idellus). GCRV is a non-enveloped, double-stranded RNA virus in the genus Aquareovirus, of the family Reoviridae, which encodes seven structural proteins (VP1-VP7) and five nonstructural proteins (NS80, NS38, NS31, NS26, and NS16). To date, the mechanism of GCRV entry into CIK Ctenopharyngon idellus kidney (CIK) cells remains poorly understood. RESULTS: Here, we present a study of the GCRV internalization mechanism in CIK cells. Our results indicated that GCRV infection was inhibited by chlorpromazine, the specific inhibitor for clathrin-mediated endocytosis. Colocalization of GCRV virions with endogenous clathrin was observed during early infection by confocal microscopy. Moreover, GCRV infection of CIK cells depended on acidification of the endosome. This was indicated by significant inhibition of viral infection following prophylactic treatment with the lysosomotropic drugs chloroquine or ammonium chloride. In addition, the disturbance of dynamin activity blocked GCRV entry, which confirmed the dynamin-dependent nature of clathrin-mediated endocytosis. CONCLUSION: Our findings suggest that GCRV might enter CIK cells via clathrin-mediated endocytosis in a pH-dependent manner. Additionally, dynamin is critical for efficient viral entry.
[Mh] Termos MeSH primário: Clatrina/metabolismo
Reoviridae/fisiologia
Internalização do Vírus
[Mh] Termos MeSH secundário: Animais
Antivirais/farmacologia
Linhagem Celular
Dinaminas/metabolismo
Inibidores Enzimáticos/farmacologia
Filipina/farmacologia
Concentração de Íons de Hidrogênio
Transporte Proteico
Reoviridae/ultraestrutura
Infecções por Reoviridae/metabolismo
Infecções por Reoviridae/virologia
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (Antiviral Agents); 0 (Clathrin); 0 (Enzyme Inhibitors); 87Z59R7D14 (Filipin); EC 3.6.5.5 (Dynamins)
[Em] Mês de entrada:1610
[Cu] Atualização por classe:161230
[Lr] Data última revisão:
161230
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:160217
[St] Status:MEDLINE
[do] DOI:10.1186/s12985-016-0485-7


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[PMID]:26801962
[Au] Autor:El-Mounadi K; Islam KT; Hernández-Ortiz P; Read ND; Shah DM
[Ad] Endereço:Donald Danforth Plant Science Center, St Louis, MO, 63132, USA.
[Ti] Título:Antifungal mechanisms of a plant defensin MtDef4 are not conserved between the ascomycete fungi Neurospora crassa and Fusarium graminearum.
[So] Source:Mol Microbiol;100(3):542-59, 2016 May.
[Is] ISSN:1365-2958
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Defensins play an important role in plant defense against fungal pathogens. The plant defensin, MtDef4, inhibits growth of the ascomycete fungi, Neurospora crassa and Fusarium graminearum, at micromolar concentrations. We have reported that MtDef4 is transported into the cytoplasm of these fungi and exerts its antifungal activity on intracellular targets. Here, we have investigated whether the antifungal mechanisms of MtDef4 are conserved in these fungi. We show that N. crassa and F. graminearum respond differently to MtDef4 challenge. Membrane permeabilization is required for the antifungal activity of MtDef4 against F. graminearum but not against N. crassa. We find that MtDef4 is targeted to different subcellular compartments in each fungus. Internalization of MtDef4 in N. crassa is energy-dependent and involves endocytosis. By contrast, MtDef4 appears to translocate into F. graminearum autonomously using a partially energy-dependent pathway. MtDef4 has been shown to bind to the phospholipid phosphatidic acid (PA). We provide evidence that the plasma membrane localized phospholipase D, involved in the biosynthesis of PA, is needed for entry of this defensin in N. crassa, but not in F. graminearum. To our knowledge, this is the first example of a defensin which inhibits the growth of two ascomycete fungi via different mechanisms.
[Mh] Termos MeSH primário: Antifúngicos/metabolismo
Defensinas/metabolismo
Endocitose/fisiologia
Fusarium/crescimento & desenvolvimento
Neurospora crassa/crescimento & desenvolvimento
[Mh] Termos MeSH secundário: Transporte Biológico Ativo/efeitos dos fármacos
Transporte Biológico Ativo/fisiologia
Brefeldina A/farmacologia
Permeabilidade da Membrana Celular/fisiologia
Endocitose/efeitos dos fármacos
Filipina/farmacologia
Ácidos Fosfatídicos/química
Fosfolipase D/química
Fosfolipase D/genética
Doenças das Plantas/imunologia
Doenças das Plantas/microbiologia
Proteínas de Plantas/metabolismo
Plantas/microbiologia
Esporos Fúngicos/efeitos dos fármacos
Esporos Fúngicos/crescimento & desenvolvimento
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Antifungal Agents); 0 (Defensins); 0 (Phosphatidic Acids); 0 (Plant Proteins); 20350-15-6 (Brefeldin A); 87Z59R7D14 (Filipin); EC 3.1.4.4 (Phospholipase D); EC 3.1.4.4 (phospholipase D1)
[Em] Mês de entrada:1707
[Cu] Atualização por classe:170713
[Lr] Data última revisão:
170713
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:160124
[St] Status:MEDLINE
[do] DOI:10.1111/mmi.13333


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[PMID]:26246267
[Au] Autor:Payero TD; Vicente CM; Rumbero Á; Barreales EG; Santos-Aberturas J; de Pedro A; Aparicio JF
[Ad] Endereço:Area of Microbiology, Faculty of Biology, Universidad de León, 24071, León, Spain. tdiep@unileon.es.
[Ti] Título:Functional analysis of filipin tailoring genes from Streptomyces filipinensis reveals alternative routes in filipin III biosynthesis and yields bioactive derivatives.
[So] Source:Microb Cell Fact;14:114, 2015 Aug 07.
[Is] ISSN:1475-2859
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:BACKGROUND: Streptomyces filipinensis is the industrial producer of filipin, a pentaene macrolide, archetype of non-glycosylated polyenes, and widely used for the detection and the quantitation of cholesterol in biological membranes and as a tool for the diagnosis of Niemann-Pick type C disease. Genetic manipulations of polyene biosynthetic pathways have proven useful for the discovery of products with improved properties. Here, we describe the late biosynthetic steps for filipin III biosynthesis and strategies for the generation of bioactive filipin III derivatives at high yield. RESULTS: A region of 13,778 base pairs of DNA from the S. filipinensis genome was isolated, sequenced, and characterized. Nine complete genes and two truncated ORFs were located. Disruption of genes proved that this genomic region is part of the biosynthetic cluster for the 28-membered ring of the polyene macrolide filipin. This set of genes includes two cytochrome P450 monooxygenase encoding genes, filC and filD, which are proposed to catalyse specific hydroxylations of the macrolide ring at C26 and C1' respectively. Gene deletion and complementation experiments provided evidence for their role during filipin III biosynthesis. Filipin III derivatives were accumulated by the recombinant mutants at high yield. These have been characterized by mass spectrometry and nuclear magnetic resonance following high-performance liquid chromatography purification thus revealing the post-polyketide steps during polyene biosynthesis. Two alternative routes lead to the formation of filipin III from the initial product of polyketide synthase chain assembly and cyclization filipin I, one trough filipin II, and the other one trough 1'-hydroxyfilipin I, all filipin III intermediates being biologically active. Moreover, minimal inhibitory concentration values against Candida utilis and Saccharomyces cerevisiae were obtained for all filipin derivatives, finding that 1'-hydroxyfilipin and especially filipin II show remarkably enhanced antifungal bioactivity. Complete nuclear magnetic resonance assignments have been obtained for the first time for 1'-hydroxyfilipin I. CONCLUSIONS: This report reveals the existence of two alternative routes for filipin III formation and opens new possibilities for the generation of biologically active filipin derivatives at high yield and with improved properties.
[Mh] Termos MeSH primário: Antibacterianos/biossíntese
Proteínas de Bactérias/genética
Sistema Enzimático do Citocromo P-450/genética
Filipina/biossíntese
Streptomyces/genética
[Mh] Termos MeSH secundário: Antibacterianos/química
Proteínas de Bactérias/metabolismo
Vias Biossintéticas
Sistema Enzimático do Citocromo P-450/metabolismo
Filipina/análogos & derivados
Dados de Sequência Molecular
Streptomyces/enzimologia
Streptomyces/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (Anti-Bacterial Agents); 0 (Bacterial Proteins); 87Z59R7D14 (Filipin); 9035-51-2 (Cytochrome P-450 Enzyme System)
[Em] Mês de entrada:1603
[Cu] Atualização por classe:170220
[Lr] Data última revisão:
170220
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:150807
[St] Status:MEDLINE
[do] DOI:10.1186/s12934-015-0307-4



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