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Pesquisa : E02.319.267.530.620.570.100 [Categoria DeCS]
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  1 / 1020 MEDLINE  
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[PMID]:28255554
[Au] Autor:Akbar S; Tahir M; Wang MB; Liu Q
[Ad] Endereço:Atta-ur-Rahman School of Applied Biosciences (ASAB), National University of Sciences and Technology (NUST), Islamabad 44000, Pakistan.
[Ti] Título:Expression Analysis of Hairpin RNA Carrying (SCMV) Derived Sequences and Transgenic Resistance Development in a Model Rice Plant.
[So] Source:Biomed Res Int;2017:1646140, 2017.
[Is] ISSN:2314-6141
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Developing transgenic resistance in monocotyledonous crops against pathogens remains a challenging area of research. (SCMV) is a serious pathogen of many monocotyledonous crops including sugarcane. The objective of present study was to analyze transgenic expression of hairpin RNA (hpRNA), targeting simultaneously (Coat Protein) and (helper component-proteinase) genes of SCMV, in a model rice plant. Conserved nucleotide sequences, exclusive for DAG (Aspartic acid-Alanine-Glycine) and KITC (Lycine-Isoleucine-Threonine-Cysteine) motifs, derived from SCMV and genes, respectively, were fused together and assembled into the hpRNA cassette under maize ubiquitin promoter to form Ubi-hpCP:Hc-Pro construct. The same CP:Hc-Pro sequence was fused with the -glucuronidase gene (GUS) at the 3' end under CaMV 35S promoter to develop 35S-GUS:CP:Hc-Pro served as a target reporter gene construct. When delivered into rice callus tissues by particle bombardment, the Ubi-hpCP:Hc-Pro construct induced strong silencing of 35S-GUS:CP:Hc-Pro. Transgenic rice plants, containing Ubi-hpCP:Hc-Pro construct, expressed high level of 21-24 nt small interfering RNAs, which induced specific suppression against GUS:CP:Hc-Pro delivered by particle bombardment and conferred strong resistance to mechanically inoculated SCMV. It is concluded that fusion hpRNA approach is an affordable method for developing resistance against SCMV in model rice plant and it could confer SCMV resistance when transformed into sugarcane.
[Mh] Termos MeSH primário: Resistência à Doença/genética
Expressão Gênica
Conformação de Ácido Nucleico
Oryza/imunologia
Oryza/virologia
Doenças das Plantas/virologia
Potyvirus/genética
RNA Viral/genética
[Mh] Termos MeSH secundário: Sequência de Bases
Biolística
Southern Blotting
Cisteína Endopeptidases/genética
Inativação Gênica
Plantas Geneticamente Modificadas
RNA Interferente Pequeno/metabolismo
RNA Viral/química
Transgenes
Proteínas Virais/genética
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (RNA, Small Interfering); 0 (RNA, Viral); 0 (Viral Proteins); EC 3.4.22.- (Cysteine Endopeptidases); EC 3.4.22.- (HC-Pro protein, potyvirus)
[Em] Mês de entrada:1703
[Cu] Atualização por classe:170313
[Lr] Data última revisão:
170313
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170304
[St] Status:MEDLINE
[do] DOI:10.1155/2017/1646140


  2 / 1020 MEDLINE  
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[PMID]:27973750
[Au] Autor:Deodhar GV; Adams ML; Trewyn BG
[Ad] Endereço:Department of Chemistry, Colorado School of Mines, Golden, CO, USA.
[Ti] Título:Controlled release and intracellular protein delivery from mesoporous silica nanoparticles.
[So] Source:Biotechnol J;12(1), 2017 Jan.
[Is] ISSN:1860-7314
[Cp] País de publicação:Germany
[La] Idioma:eng
[Ab] Resumo:Protein therapeutics are promising candidates for disease treatment due to their high specificity and minimal adverse side effects; however, targeted protein delivery to specific sites has proven challenging. Mesoporous silica nanoparticles (MSN) have demonstrated to be ideal candidates for this application, given their high loading capacity, biocompatibility, and ability to protect host molecules from degradation. These materials exhibit tunable pore sizes, shapes and volumes, and surfaces which can be easily functionalized. This serves to control the movement of molecules in and out of the pores, thus entrapping guest molecules until a specific stimulus triggers release. In this review, we will cover the benefits of using MSN as protein therapeutic carriers, demonstrating that there is great diversity in the ways MSN can be used to service proteins. Methods for controlling the physical dimensions of pores via synthetic conditions, applications of therapeutic protein loaded MSN materials in cancer therapies, delivering protein loaded MSN materials to plant cells using biolistic methods, and common stimuli-responsive functionalities will be discussed. New and exciting strategies for controlled release and manipulation of proteins are also covered in this review. While research in this area has advanced substantially, we conclude this review with future challenges to be tackled by the scientific community.
[Mh] Termos MeSH primário: Sistemas de Liberação de Medicamentos/métodos
Nanopartículas
Proteínas/administração & dosagem
[Mh] Termos MeSH secundário: Animais
Biolística/métodos
Preparações de Ação Retardada
Portadores de Fármacos
Seres Humanos
Concentração de Íons de Hidrogênio
Magnetismo
Nanopartículas/administração & dosagem
Nanopartículas/química
Porosidade
Proteínas/química
Proteínas/farmacocinética
Dióxido de Silício
[Pt] Tipo de publicação:JOURNAL ARTICLE; REVIEW
[Nm] Nome de substância:
0 (Delayed-Action Preparations); 0 (Drug Carriers); 0 (Proteins); 7631-86-9 (Silicon Dioxide)
[Em] Mês de entrada:1701
[Cu] Atualização por classe:170123
[Lr] Data última revisão:
170123
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:161216
[St] Status:MEDLINE
[do] DOI:10.1002/biot.201600408


  3 / 1020 MEDLINE  
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[PMID]:27826796
[Au] Autor:Chee MJ; Lycett GW; Khoo TJ; Chin CF
[Ad] Endereço:Faculty of Science, School of Biosciences, University of Nottingham, Malaysia Campus, 43500, Semenyih, Selangor Darul Ehsan, Malaysia.
[Ti] Título:Bioengineering of the Plant Culture of Capsicum frutescens with Vanillin Synthase Gene for the Production of Vanillin.
[So] Source:Mol Biotechnol;59(1):1-8, 2017 Jan.
[Is] ISSN:1559-0305
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Production of vanillin by bioengineering has gained popularity due to consumer demand toward vanillin produced by biological systems. Natural vanillin from vanilla beans is very expensive to produce compared to its synthetic counterpart. Current bioengineering works mainly involve microbial biotechnology. Therefore, alternative means to the current approaches are constantly being explored. This work describes the use of vanillin synthase (VpVAN), to bioconvert ferulic acid to vanillin in a plant system. The VpVAN enzyme had been shown to directly convert ferulic acid and its glucoside into vanillin and its glucoside, respectively. As the ferulic acid precursor and vanillin were found to be the intermediates in the phenylpropanoid biosynthetic pathway of Capsicum species, this work serves as a proof-of-concept for vanillin production using Capsicum frutescens (C. frutescens or hot chili pepper). The cells of C. frutescens were genetically transformed with a codon optimized VpVAN gene via biolistics. Transformed explants were selected and regenerated into callus. Successful integration of the gene cassette into the plant genome was confirmed by polymerase chain reaction. High-performance liquid chromatography was used to quantify the phenolic compounds detected in the callus tissues. The vanillin content of transformed calli was 0.057% compared to 0.0003% in untransformed calli.
[Mh] Termos MeSH primário: Benzaldeídos/metabolismo
Biolística/métodos
Capsicum/crescimento & desenvolvimento
Hidroliases/metabolismo
[Mh] Termos MeSH secundário: Bioengenharia/métodos
Vias Biossintéticas
Capsicum/genética
Ácidos Cumáricos/metabolismo
Hidroliases/genética
Proteínas de Plantas/genética
Proteínas de Plantas/metabolismo
Plantas Geneticamente Modificadas/crescimento & desenvolvimento
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Benzaldehydes); 0 (Coumaric Acids); 0 (Plant Proteins); AVM951ZWST (ferulic acid); CHI530446X (vanillin); EC 4.2.1.- (Hydro-Lyases)
[Em] Mês de entrada:1705
[Cu] Atualização por classe:171108
[Lr] Data última revisão:
171108
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:161110
[St] Status:MEDLINE
[do] DOI:10.1007/s12033-016-9986-2


  4 / 1020 MEDLINE  
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[PMID]:27796719
[Au] Autor:Zienkiewicz M; Krupnik T; Drozak A; Golke A; Romanowska E
[Ad] Endereço:Department of Molecular Plant Physiology, Faculty of Biology, University of Warsaw, ul. Miecznikowa 1, 02-096, Warsaw, Poland. maximus@biol.uw.edu.pl.
[Ti] Título:Transformation of the Cyanidioschyzon merolae chloroplast genome: prospects for understanding chloroplast function in extreme environments.
[So] Source:Plant Mol Biol;93(1-2):171-183, 2017 Jan.
[Is] ISSN:1573-5028
[Cp] País de publicação:Netherlands
[La] Idioma:eng
[Ab] Resumo:KEY MESSAGE: We have successfully transformed an exthemophilic red alga with the chloramphenicol acetyltransferase gene, rendering this organism insensitive to its toxicity. Our work paves the way to further work with this new modelorganism. Here we report the first successful attempt to achieve a stable, under selectable pressure, chloroplast transformation in Cyanidioschizon merolae-an extremophilic red alga of increasing importance as a new model organism. The following protocol takes advantage of a double homologous recombination phenomenon in the chloroplast, allowing to introduce an exogenous, selectable gene. For that purpose, we decided to use chloramphenicol acetyltransferase (CAT), as chloroplasts are particularly vulnerable to chloramphenicol lethal effects (Zienkiewicz et al. in Protoplasma, 2015, doi: 10.1007/s00709-015-0936-9 ). We adjusted two methods of DNA delivery: the PEG-mediated delivery and the biolistic bombardment based delivery, either of these methods work sufficiently with noticeable preference to the former. Application of a codon-optimized sequence of the cat gene and a single colony selection yielded C. merolae strains, capable of resisting up to 400 µg/mL of chloramphenicol. Our method opens new possibilities in production of site-directed mutants, recombinant proteins and exogenous protein overexpression in C. merolae-a new model organism.
[Mh] Termos MeSH primário: Cloroplastos/genética
Genoma de Cloroplastos
Rodófitas/genética
[Mh] Termos MeSH secundário: Biolística
Cloranfenicol O-Acetiltransferase/genética
Cloroplastos/fisiologia
Recombinação Homóloga
Plantas Geneticamente Modificadas
Transformação Genética
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
EC 2.3.1.28 (Chloramphenicol O-Acetyltransferase)
[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:161101
[St] Status:MEDLINE
[do] DOI:10.1007/s11103-016-0554-8


  5 / 1020 MEDLINE  
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[PMID]:27787577
[Au] Autor:Eby JM; Barse L; Henning SW; Rabelink MJ; Klarquist J; Gilbert ER; Hammer AM; Fernandez MF; Yung N; Khan S; Miller HG; Kessler ER; Garrett-Mayer E; Dilling DF; Hoeben RC; Le Poole IC
[Ad] Endereço:Oncology Research Institute, Loyola University Medical Center, Loyola University Chicago, Rm 203, 2160 S. 1st Avenue, Maywood, IL, 60153, USA.
[Ti] Título:Alpha-N-acetyl-neuraminide alpha-2,8-sialyltransferase 1 can support immune responses toward tumors overexpressing ganglioside D3 in mice.
[So] Source:Cancer Immunol Immunother;66(1):63-75, 2017 Jan.
[Is] ISSN:1432-0851
[Cp] País de publicação:Germany
[La] Idioma:eng
[Ab] Resumo:An immunotherapeutic strategy is discussed supporting anti-tumor activity toward malignancies overexpressing ganglioside D3. GD3 can be targeted by NKT cells when derived moieties are presented in the context of CD1d. NKT cells can support anti-tumor responses by secreting inflammatory cytokines and through cytotoxicity toward CD1d GD3 tumors. To overexpress GD3, we generated expression vector DNA and an adenoviral vector encoding the enzyme responsible for generating GD3 from its ubiquitous precursor GM3. We show that DNA encoding α-N-acetyl-neuraminide α-2,8-sialyltransferase 1 (SIAT8) introduced by gene gun vaccination in vivo leads to overexpression of GD3 and delays tumor growth. Delayed tumor growth is dependent on CD1d expression by host immune cells, as shown in experiments engaging CD1d knockout mice. A trend toward greater NKT cell populations among tumor-infiltrating lymphocytes is associated with SIAT8 vaccination. A single adenoviral vaccination introduces anti-tumor activity similarly to repeated vaccination with naked DNA. Here, greater NKT tumor infiltrates were accompanied by marked overexpression of IL-17 in the tumor, later switching to IL-4. Our results suggest that a single intramuscular adenoviral vaccination introduces overexpression of GD3 by antigen-presenting cells at the injection site, recruiting NKT cells that provide an inflammatory anti-tumor environment. We propose adenoviral SIAT8 (AdV-SIAT8) can slow the growth of GD3 expressing tumors in patients.
[Mh] Termos MeSH primário: Gangliosídeos/biossíntese
Melanoma Experimental/imunologia
Melanoma/imunologia
Sialiltransferases/imunologia
[Mh] Termos MeSH secundário: Animais
Biolística
Linhagem Celular Tumoral
Gangliosídeos/imunologia
Células HEK293
Seres Humanos
Melanoma/enzimologia
Melanoma/terapia
Melanoma Experimental/enzimologia
Melanoma Experimental/terapia
Camundongos
Camundongos Endogâmicos C57BL
Camundongos Knockout
Sialiltransferases/genética
Vacinas de DNA/imunologia
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Gangliosides); 0 (Vaccines, DNA); 62010-37-1 (ganglioside, GD3); EC 2.4.99.- (Sialyltransferases); EC 2.4.99.8 (alpha-N-acetylneuraminate alpha-2,8-sialyltransferase); EC 3.4.99.- (ST8SIA5 protein, human)
[Em] Mês de entrada:1709
[Cu] Atualização por classe:170919
[Lr] Data última revisão:
170919
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:161028
[St] Status:MEDLINE
[do] DOI:10.1007/s00262-016-1920-8


  6 / 1020 MEDLINE  
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[PMID]:27277128
[Au] Autor:Wu H; Acanda Y; Jia H; Wang N; Zale J
[Ad] Endereço:Plant Pathology Department, Institute of Food and Agricultural Sciences, Citrus Research and Education Center, University of Florida, Lake Alfred, FL, 33850, USA. whao@ufl.edu.
[Ti] Título:Biolistic transformation of Carrizo citrange (Citrus sinensis Osb. × Poncirus trifoliata L. Raf.).
[So] Source:Plant Cell Rep;35(9):1955-62, 2016 Sep.
[Is] ISSN:1432-203X
[Cp] País de publicação:Germany
[La] Idioma:eng
[Ab] Resumo:KEY MESSAGE: The development of transgenic citrus plants by the biolistic method. A protocol for the biolistic transformation of epicotyl explants and transgenic shoot regeneration of immature citrange rootstock, cv. Carrizo (Citrus sinensis Osb. × Poncirus trifoliata L. Raf.) and plant regeneration is described. Immature epicotyl explants were bombarded with a vector containing the nptII selectable marker and the gfp reporter. The number of independent, stably transformed tissues/total number of explants, recorded by monitoring GFP fluorescence 4 weeks after bombardment was substantial at 18.4 %, and some fluorescing tissues regenerated into shoots. Fluorescing GFP, putative transgenic shoots were micro-grafted onto immature Carrizo rootstocks in vitro, confirmed by PCR amplification of nptII and gfp coding regions, followed by secondary grafting onto older rootstocks grown in soil. Southern blot analysis indicated that all the fluorescing shoots were transgenic. Multiple and single copies of nptII integrations were confirmed in five regenerated transgenic lines. There is potential to develop a higher throughput biolistics transformation system by optimizing the tissue culture medium to improve shoot regeneration and narrowing the window for plant sampling. This system will be appropriate for transformation with minimal cassettes.
[Mh] Termos MeSH primário: Biolística/métodos
Citrus sinensis/genética
Cruzamentos Genéticos
Poncirus/genética
Transformação Genética
[Mh] Termos MeSH secundário: Southern Blotting
Fenótipo
Brotos de Planta/genética
Plantas Geneticamente Modificadas
Plântulas/genética
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Em] Mês de entrada:1703
[Cu] Atualização por classe:171004
[Lr] Data última revisão:
171004
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:160610
[St] Status:MEDLINE
[do] DOI:10.1007/s00299-016-2010-2


  7 / 1020 MEDLINE  
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[PMID]:27130499
[Au] Autor:Aps LRMM; Tavares MB; Rozenfeld JHK; Lamy MT; Ferreira LCS; Diniz MO
[Ad] Endereço:Vaccine Development Laboratory, Institute of Biomedical Sciences, University of São Paulo, São Paulo, Brazil.
[Ti] Título:Bacterial spores as particulate carriers for gene gun delivery of plasmid DNA.
[So] Source:J Biotechnol;228:58-66, 2016 Jun 20.
[Is] ISSN:1873-4863
[Cp] País de publicação:Netherlands
[La] Idioma:eng
[Ab] Resumo:Bacillus subtilis spores represent a suitable platform for the adsorption of proteins, enzymes and viral particles at physiological conditions. In the present work, we demonstrate that purified spores can also adsorb DNA on their surface after treatment with cationic molecules. In addition, we demonstrate that DNA-coated B. subtilis spores can be used as particulate carriers and act as an alternative to gold microparticles for the biolistic (gene gun) administration of plasmid DNA in mice. Gene gun delivery of spores pre-treated with DODAB (dioctadecyldimethylammonium bromide) allowed efficient plasmid DNA absorption and induced protein expression levels similar to those obtained with gold microparticles. More importantly, we demonstrated that a DNA vaccine adsorbed on spores can be loaded into biolistic cartridges and efficiently delivered into mice, which induced specific cellular and antibody responses. Altogether, these data indicate that B. subtilis spores represent a simple and low cost alternative for the in vivo delivery of DNA vaccines by the gene gun technology.
[Mh] Termos MeSH primário: Biolística/métodos
Portadores de Fármacos/química
Esporos Bacterianos/química
Vacinas de DNA/química
[Mh] Termos MeSH secundário: Adsorção
Animais
Bacillus subtilis/química
Portadores de Fármacos/administração & dosagem
Ouro/química
Masculino
Camundongos
Camundongos Endogâmicos C57BL
Proteínas Oncogênicas Virais/genética
Proteínas Oncogênicas Virais/imunologia
Compostos de Amônio Quaternário/química
Esporos Bacterianos/imunologia
Vacinas de DNA/administração & dosagem
Vacinas de DNA/imunologia
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Drug Carriers); 0 (Oncogene Proteins, Viral); 0 (Quaternary Ammonium Compounds); 0 (Vaccines, DNA); 251IW5I21C (dimethyldioctadecylammonium); 7440-57-5 (Gold)
[Em] Mês de entrada:1702
[Cu] Atualização por classe:170806
[Lr] Data última revisão:
170806
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:160501
[St] Status:MEDLINE


  8 / 1020 MEDLINE  
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[PMID]:27085173
[Au] Autor:McBurney SP; Sunshine JE; Gabriel S; Huynh JP; Sutton WF; Fuller DH; Haigwood NL; Messer WB
[Ad] Endereço:Division of Pathobiology & Immunology, Oregon National Primate Research Center, Oregon Health & Science University, 505 NW 185th Ave., Beaverton, OR 97006, USA.
[Ti] Título:Evaluation of protection induced by a dengue virus serotype 2 envelope domain III protein scaffold/DNA vaccine in non-human primates.
[So] Source:Vaccine;34(30):3500-7, 2016 Jun 24.
[Is] ISSN:1873-2518
[Cp] País de publicação:Netherlands
[La] Idioma:eng
[Ab] Resumo:We describe the preclinical development of a dengue virus vaccine targeting the dengue virus serotype 2 (DENV2) envelope domain III (EDIII). This study provides proof-of-principle that a dengue EDIII protein scaffold/DNA vaccine can protect against dengue challenge. The dengue vaccine (EDIII-E2) is composed of both a protein particle and a DNA expression plasmid delivered simultaneously via intramuscular injection (protein) and gene gun (DNA) into rhesus macaques. The protein component can contain a maximum of 60 copies of EDIII presented on a multimeric scaffold of Geobacillus stearothermophilus E2 proteins. The DNA component is composed of the EDIII portion of the envelope gene cloned into an expression plasmid. The EDIII-E2 vaccine elicited robust antibody responses to DENV2, with neutralizing antibody responses detectable following the first boost and reaching titers of greater than 1:100,000 following the second and final boost. Vaccinated and naïve groups of macaques were challenged with DENV2. All vaccinated macaques were protected from detectable viremia by infectious assay, while naïve animals had detectable viremia for 2-7 days post-challenge. All naïve macaques had detectable viral RNA from day 2-10 post-challenge. In the EDIII-E2 group, three macaques were negative for viral RNA and three were found to have detectable viral RNA post challenge. Viremia onset was delayed and the duration was shortened relative to naïve controls. The presence of viral RNA post-challenge corresponded to a 10-30-fold boost in neutralization titers 28 days post challenge, whereas no boost was observed in the fully protected animals. Based on these results, we determine that pre-challenge 50% neutralization titers of >1:6000 correlated with sterilizing protection against DENV2 challenge in EDIII-E2 vaccinated macaques. Identification of the critical correlate of protection for the EDIII-E2 platform in the robust non-human primate model lays the groundwork for further development of a tetravalent EDIII-E2 dengue vaccine.
[Mh] Termos MeSH primário: Vacinas contra Dengue/imunologia
Vírus da Dengue/classificação
Dengue/prevenção & controle
Vacinas de DNA/imunologia
[Mh] Termos MeSH secundário: Animais
Anticorpos Neutralizantes/sangue
Anticorpos Antivirais/sangue
Biolística
Macaca mulatta
RNA Viral/sangue
Proteínas Recombinantes de Fusão/imunologia
Vacinas Sintéticas/imunologia
Viremia/prevenção & controle
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Antibodies, Neutralizing); 0 (Antibodies, Viral); 0 (Dengue Vaccines); 0 (RNA, Viral); 0 (Recombinant Fusion Proteins); 0 (Vaccines, DNA); 0 (Vaccines, Synthetic)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171026
[Lr] Data última revisão:
171026
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:160417
[St] Status:MEDLINE


  9 / 1020 MEDLINE  
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[PMID]:26908053
[Au] Autor:Hansen DT; Robida MD; Craciunescu FM; Loskutov AV; Dörner K; Rodenberry JC; Wang X; Olson TL; Patel H; Fromme P; Sykes KF
[Ad] Endereço:Center for Innovations in Medicine, Biodesign Institute, Arizona State University, Tempe, Arizona, USA.
[Ti] Título:Polyclonal Antibody Production for Membrane Proteins via Genetic Immunization.
[So] Source:Sci Rep;6:21925, 2016 Feb 24.
[Is] ISSN:2045-2322
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Antibodies are essential for structural determinations and functional studies of membrane proteins, but antibody generation is limited by the availability of properly-folded and purified antigen. We describe the first application of genetic immunization to a structurally diverse set of membrane proteins to show that immunization of mice with DNA alone produced antibodies against 71% (n = 17) of the bacterial and viral targets. Antibody production correlated with prior reports of target immunogenicity in host organisms, underscoring the efficiency of this DNA-gold micronanoplex approach. To generate each antigen for antibody characterization, we also developed a simple in vitro membrane protein expression and capture method. Antibody specificity was demonstrated upon identifying, for the first time, membrane-directed heterologous expression of the native sequences of the FopA and FTT1525 virulence determinants from the select agent Francisella tularensis SCHU S4. These approaches will accelerate future structural and functional investigations of therapeutically-relevant membrane proteins.
[Mh] Termos MeSH primário: Anticorpos/isolamento & purificação
Proteínas da Membrana Bacteriana Externa/imunologia
DNA Bacteriano/imunologia
Francisella tularensis/imunologia
Imunoconjugados/administração & dosagem
Tularemia/prevenção & controle
Fatores de Virulência/imunologia
[Mh] Termos MeSH secundário: Animais
Anticorpos/metabolismo
Especificidade de Anticorpos
Antígenos de Bactérias/genética
Antígenos de Bactérias/imunologia
Proteínas da Membrana Bacteriana Externa/genética
Biolística
DNA Bacteriano/genética
Feminino
Francisella tularensis/genética
Francisella tularensis/patogenicidade
Regulação da Expressão Gênica
Vetores Genéticos/administração & dosagem
Vetores Genéticos/química
Vetores Genéticos/metabolismo
Ouro/química
Proteínas de Fluorescência Verde/genética
Proteínas de Fluorescência Verde/imunologia
Imunização/instrumentação
Imunização/métodos
Imunoconjugados/genética
Nanopartículas de Magnetita/química
Camundongos
Camundongos Endogâmicos BALB C
Biossíntese de Proteínas
Proteínas Recombinantes de Fusão/genética
Proteínas Recombinantes de Fusão/imunologia
Tularemia/imunologia
Tularemia/microbiologia
Fatores de Virulência/genética
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, N.I.H., EXTRAMURAL
[Nm] Nome de substância:
0 (Antibodies); 0 (Antigens, Bacterial); 0 (Bacterial Outer Membrane Proteins); 0 (DNA, Bacterial); 0 (FopA protein, Francisella tularensis); 0 (Immunoconjugates); 0 (Magnetite Nanoparticles); 0 (Recombinant Fusion Proteins); 0 (Virulence Factors); 147336-22-9 (Green Fluorescent Proteins); 7440-57-5 (Gold)
[Em] Mês de entrada:1612
[Cu] Atualização por classe:170220
[Lr] Data última revisão:
170220
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:160225
[St] Status:MEDLINE
[do] DOI:10.1038/srep21925


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[PMID]:26764230
[Au] Autor:Tsai SW; Tung YT; Chen HL; Yang SH; Liu CY; Lu M; Pai HJ; Lin CC; Chen CM
[Ad] Endereço:Department of Life Sciences, Agricultural Biotechnology Center, National Chung Hsing University, Taichung 402, Taiwan; Department of Physical Medicine and Rehabilitation, Taichung Tzu Chi Hospital, Buddhist Tzu Chi Medical Foundation, Taichung 404, Taiwan; Department of Physical Medicine and Rehabil
[Ti] Título:Myostatin propeptide gene delivery by gene gun ameliorates muscle atrophy in a rat model of botulinum toxin-induced nerve denervation.
[So] Source:Life Sci;146:15-23, 2016 Feb 01.
[Is] ISSN:1879-0631
[Cp] País de publicação:Netherlands
[La] Idioma:eng
[Ab] Resumo:AIM: Muscle atrophy is a common symptom after nerve denervation. Myostatin propeptide, a precursor of myostatin, has been documented to improve muscle growth. However, the mechanism underlying the muscle atrophy attenuation effects of myostatin propeptide in muscles and the changes in gene expression are not well established. We investigated the possible underlying mechanisms associated with myostatin propeptide gene delivery by gene gun in a rat denervation muscle atrophy model, and evaluated gene expression patterns. MAIN METHODS: In a rat botulinum toxin-induced nerve denervation muscle atrophy model, we evaluated the effects of wild-type (MSPP) and mutant-type (MSPPD75A) of myostatin propeptide gene delivery, and observed changes in gene activation associated with the neuromuscular junction, muscle and nerve. KEY FINDINGS: Muscle mass and muscle fiber size was moderately increased in myostatin propeptide treated muscles (p<0.05). And enhancement of the gene expression of the muscle regulatory factors, neurite outgrowth factors (IGF-1, GAP43) and acetylcholine receptors was observed. Our results demonstrate that myostatin propeptide gene delivery, especially the mutant-type of MSPPD75A, attenuates muscle atrophy through myogenic regulatory factors and acetylcholine receptor regulation. SIGNIFICANCE: Our data concluded that myostatin propeptide gene therapy may be a promising treatment for nerve denervation induced muscle atrophy.
[Mh] Termos MeSH primário: Biolística/métodos
Toxinas Botulínicas
Atrofia Muscular/terapia
Miostatina/genética
[Mh] Termos MeSH secundário: Animais
Expressão Gênica/efeitos dos fármacos
Técnicas de Transferência de Genes
Denervação Muscular
Desenvolvimento Muscular/efeitos dos fármacos
Fibras Musculares Esqueléticas/metabolismo
Músculo Esquelético/inervação
Músculo Esquelético/metabolismo
Atrofia Muscular/induzido quimicamente
Atrofia Muscular/patologia
Regeneração Nervosa
Junção Neuromuscular/metabolismo
Plasmídeos/genética
Ratos
Ratos Sprague-Dawley
Receptores Colinérgicos/biossíntese
Receptores Colinérgicos/genética
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (Gdf8 protein, rat); 0 (Myostatin); 0 (Receptors, Cholinergic); EC 3.4.24.69 (Botulinum Toxins)
[Em] Mês de entrada:1606
[Cu] Atualização por classe:160208
[Lr] Data última revisão:
160208
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:160115
[St] Status:MEDLINE



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