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[PMID]:29458684
[Au] Autor:Wise MG; Horvath E; Young K; Sahm DF; Kazmierczak KM
[Ad] Endereço:1​International Health Management Associates, Schaumburg, Illinois, USA.
[Ti] Título:Global survey of Klebsiella pneumoniae major porins from ertapenem non-susceptible isolates lacking carbapenemases.
[So] Source:J Med Microbiol;67(3):289-295, 2018 Mar.
[Is] ISSN:1473-5644
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:PURPOSE: To understand the diversity of porin disruption in Klebsiella pneumoniae, the major outer membrane protein (OMP) porins, OmpK35 and OmpK36, were examined in a set of isolates that did not harbour traditional carbapenem-hydrolysing enzymes, but nevertheless tested non-susceptible to ertapenem. METHODS: A world-wide collection of Klebsiella pneumoniae isolates that were part of the Study for Monitoring Antimicrobial Resistance Trends (SMART) surveillance project over the years 2008-2014 were characterised with regard to their ß-lactamase gene carriage and potential permeability defects. Four hundred and eighty-seven isolates that did not carry carbapenemase genes, but were non-susceptible to ertapenem, were investigated by sequence analysis of the genes encoding OmpK35 and OmpK36. Isolates without obvious genetic lesions in either major porin gene were further examined by outer membrane protein SDS-PAGE. RESULTS: The majority of isolates, 83.0 % (404/487), exhibited clear genetic disruption in either or both of the ompK35 and ompK36 genes. Among the proportion of the collection with the highest ertapenem MIC value (>4 mg l ), 60.5 % (115/190) showed mutation in both porin genes. Isolates without obvious genetic mutations were examined by SDS-PAGE, and 90.4 % (75/83) were found to lack or show altered expression of at least one of the major OMPs when compared to an ertapenem sensitive control strain. CONCLUSION: This study illustrates that porin deficiency in Klebsiella pneumoniae is a widespread phenomenon, and in combination with ESBLs and/or AmpC enzymes, likely accounts for the elevated ertapenem MICs observed in this study.
[Mh] Termos MeSH primário: Antibacterianos/farmacologia
Proteínas de Bactérias/genética
Klebsiella pneumoniae/genética
Porinas/genética
beta-Lactamas/farmacologia
[Mh] Termos MeSH secundário: Proteínas de Bactérias/metabolismo
Carbapenêmicos/farmacologia
DNA Bacteriano/genética
Eletroforese em Gel de Poliacrilamida
Seres Humanos
Infecções por Klebsiella/epidemiologia
Infecções por Klebsiella/microbiologia
Klebsiella pneumoniae/efeitos dos fármacos
Klebsiella pneumoniae/isolamento & purificação
Klebsiella pneumoniae/metabolismo
Testes de Sensibilidade Microbiana
Mutação
beta-Lactamases/genética
beta-Lactamases/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Anti-Bacterial Agents); 0 (Bacterial Proteins); 0 (Carbapenems); 0 (DNA, Bacterial); 0 (OmpK35 porin, Klebsiella pneumoniae); 0 (OmpK36 protein, Klebsiella pneumoniae); 0 (Porins); 0 (beta-Lactams); EC 3.5.2.6 (beta-Lactamases); EC 3.5.2.6 (carbapenemase); G32F6EID2H (ertapenem)
[Em] Mês de entrada:1803
[Cu] Atualização por classe:180308
[Lr] Data última revisão:
180308
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:180221
[St] Status:MEDLINE
[do] DOI:10.1099/jmm.0.000691


  2 / 125115 MEDLINE  
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[PMID]:29267499
[Au] Autor:Santos APP; Silva MDS; Costa EVL; Rufino RD; Santos VA; Ramos CS; Sarubbo LA; Porto ALF
[Ad] Endereço:Departamento de Morfologia e Fisiologia Animal, Universidade Federal Rural de Pernambuco, Recife, PE, Brasil.
[Ti] Título:Production and characterization of a biosurfactant produced by Streptomyces sp. DPUA 1559 isolated from lichens of the Amazon region.
[So] Source:Braz J Med Biol Res;51(2):e6657, 2017 Dec 11.
[Is] ISSN:1414-431X
[Cp] País de publicação:Brazil
[La] Idioma:eng
[Ab] Resumo:Surfactants are amphipathic compounds containing both hydrophilic and hydrophobic groups, capable to lower the surface or interfacial tension. Considering the advantages of the use of biosurfactants produced by microorganisms, the aim of this paper was to develop and characterize a biosurfactant produced by Streptomyces sp. DPUA1559 isolated from lichens of the Amazon region. The microorganism was cultured in a mineral medium containing 1% residual frying soybean oil as the carbon source. The kinetics of biosurfactant production was accompanied by reducing the surface tension of the culture medium from 60 to values around 27.14 mN/m, and by the emulsification index, which showed the efficiency of the biosurfactant as an emulsifier of hydrophobic compounds. The yield of the isolated biosurfactant was 1.74 g/L, in addition to the excellent capability of reducing the surface tension (25.34 mN/m), as observed from the central composite rotational design when the biosurfactant was produced at pH 8.5 at 28°C. The critical micelle concentration of the biosurfactant was determined as 0.01 g/mL. The biosurfactant showed thermal and pH stability regarding the surface tension reduction, and tolerance under high salt concentrations. The isolated biosurfactant showed no toxicity to the micro-crustacean Artemia salina, and to the seeds of lettuce (Lactuca sativa L.) and cabbage (Brassica oleracea L.). The biochemistry characterization of the biosurfactant showed a single protein band, an acid character and a molecular weight around 14.3 kDa, suggesting its glycoproteic nature. The results are promising for the industrial application of this new biosurfactant.
[Mh] Termos MeSH primário: Líquens/microbiologia
Streptomyces/metabolismo
Tensoativos/metabolismo
[Mh] Termos MeSH secundário: Análise de Variância
Contagem de Colônia Microbiana
Meios de Cultura
Eletroforese em Gel de Poliacrilamida
Fermentação
Concentração de Íons de Hidrogênio
Valores de Referência
Sementes/efeitos dos fármacos
Óleo de Soja/química
Espectroscopia de Infravermelho com Transformada de Fourier
Streptomyces/crescimento & desenvolvimento
Streptomyces/isolamento & purificação
Tensão Superficial
Tensoativos/análise
Tensoativos/química
Temperatura Ambiente
Fatores de Tempo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Culture Media); 0 (Surface-Active Agents); 8001-22-7 (Soybean Oil)
[Em] Mês de entrada:1803
[Cu] Atualização por classe:180308
[Lr] Data última revisão:
180308
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171222
[St] Status:MEDLINE


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[PMID]:29205002
[Au] Autor:Ding DX; Ding M
[Ad] Endereço:Department of Forensic Serology, Faculty of Forensic Medicine, China Medical University, Shenyang 110001, China.
[Ti] Título:[Application of Multiple Displacement Amplification in Samples with Inhibitors].
[So] Source:Fa Yi Xue Za Zhi;32(5):342-345, 2016 Oct.
[Is] ISSN:1004-5619
[Cp] País de publicação:China
[La] Idioma:chi
[Ab] Resumo:OBJECTIVES: To explore the ability of inhibition resistibility of multiple displacement amplification (MDA) in samples with inhibitors. To explain the application and value of MDA in forensic medicine by comparing with using magnetic beads methods (MBM) to purify sample. METHODS: Different concentrations of hemoglobin and humid acid (HA) mixed with DNA samples and then divided the samples into MDA group, MBM group and control group. locus was amplified and detected by polyacrylamide gel electrophoresis detection system and AmpFâ„“STR® Identifiler™ Plus Kit-capillary electrophoresis detection system. RESULTS: When hemoglobin concentrations exceed 1 ng/µL or HA concentrations exceed 0.1 ng/µL, amplification products could not be obtained by single-locus system in control group. When hemoglobin concentration exceeds 100 ng/µL or HA concentrations exceed 1 ng/µL, the samples could not be amplified by MBM. Inhibitors in different concentrations were amplified successfully in MDA group without any influence from inhibitors. CONCLUSIONS: MDA has the capability to remove the inhibition of hemoglobin and HA, which is better than MBM and has a certain value in forensic practices.
[Mh] Termos MeSH primário: DNA/análise
Medicina Legal/métodos
Técnicas de Amplificação de Ácido Nucleico
[Mh] Termos MeSH secundário: Eletroforese em Gel de Poliacrilamida
Hemoglobinas
Seres Humanos
Substâncias Húmicas
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Hemoglobins); 0 (Humic Substances); 9007-49-2 (DNA)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180308
[Lr] Data última revisão:
180308
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171206
[St] Status:MEDLINE
[do] DOI:10.3969/j.issn.1004-5619.2016.05.006


  4 / 125115 MEDLINE  
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[PMID]:29287247
[Au] Autor:Beloborodov SS; Bao J; Krylova SM; Shala-Lawrence A; Johnson PE; Krylov SN
[Ad] Endereço:Department of Chemistry and Centre for Research on Biomolecular Interactions, York University, Toronto, Ontario M3J 1P3, Canada.
[Ti] Título:Aptamer facilitated purification of functional proteins.
[So] Source:J Chromatogr B Analyt Technol Biomed Life Sci;1073:201-206, 2018 Jan 15.
[Is] ISSN:1873-376X
[Cp] País de publicação:Netherlands
[La] Idioma:eng
[Ab] Resumo:DNA aptamers are attractive capture probes for affinity chromatography since, in contrast to antibodies, they can be chemically synthesized and, in contrast to tag-specific capture probes (such as Nickel-NTA or Glutathione), they can be used for purification of proteins free of genetic modifications (such as His or GST tags). Despite these attractive features of aptamers as capture probes, there are only a few reports on aptamer-based protein purification and none of them includes a test of the purified protein's activity, thus, leaving discouraging doubts about method's ability to purify proteins in their active state. The goal of this work was to prove that aptamers could facilitate isolation of active proteins. We refined a complete aptamer-based affinity purification procedure, which takes 4 h to complete. We further applied this procedure to purify two recombinant proteins, MutS and AlkB, from bacterial cell culture: 0.21 mg of 85%-pure AlkB from 4 mL of culture and 0.24 mg of 82%-pure MutS from 0.5 mL of culture. Finally, we proved protein activity by two capillary electrophoresis based assays: an enzymatic assay for AlkB and a DNA-binding assay for MutS. We suggest that in combination with aptamer selection for non-purified protein targets in crude cell lysate, aptamer-based purification provides a means of fast isolation of tag-free recombinant proteins in their native state without the use of antibodies.
[Mh] Termos MeSH primário: Aptâmeros de Nucleotídeos/química
Cromatografia de Afinidade/métodos
Ácidos Nucleicos Imobilizados/química
Proteínas Recombinantes/isolamento & purificação
[Mh] Termos MeSH secundário: Enzimas AlkB/química
Enzimas AlkB/genética
Enzimas AlkB/isolamento & purificação
Enzimas AlkB/metabolismo
Aptâmeros de Nucleotídeos/metabolismo
Eletroforese Capilar
Eletroforese em Gel de Poliacrilamida
Escherichia coli/genética
Ácidos Nucleicos Imobilizados/metabolismo
Metilação
Proteína MutS de Ligação de DNA com Erro de Pareamento/química
Proteína MutS de Ligação de DNA com Erro de Pareamento/genética
Proteína MutS de Ligação de DNA com Erro de Pareamento/isolamento & purificação
Proteína MutS de Ligação de DNA com Erro de Pareamento/metabolismo
Ligação Proteica
Proteínas Recombinantes/química
Proteínas Recombinantes/genética
Proteínas Recombinantes/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Aptamers, Nucleotide); 0 (Immobilized Nucleic Acids); 0 (Recombinant Proteins); EC 1.14.11.33 (AlkB Enzymes); EC 3.6.1.3 (MutS DNA Mismatch-Binding Protein)
[Em] Mês de entrada:1803
[Cu] Atualização por classe:180302
[Lr] Data última revisão:
180302
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171230
[St] Status:MEDLINE


  5 / 125115 MEDLINE  
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[PMID]:29248767
[Au] Autor:G AG; Kamalanathan AS; Vijayalakshmi MA; Venkataraman K
[Ad] Endereço:Centre for Bioseparation Technology, VIT University, Vellore- 632014, Tamil Nadu, India.
[Ti] Título:Efficient purification of Apolipoprotein A1 (ApoA1) from plasma by HEA HyperCel™: An alternative approach.
[So] Source:J Chromatogr B Analyt Technol Biomed Life Sci;1073:104-109, 2018 Jan 15.
[Is] ISSN:1873-376X
[Cp] País de publicação:Netherlands
[La] Idioma:eng
[Ab] Resumo:HDL-ApoA1 plays a pivotal role in the prevention of atherosclerosis and cardiovascular diseases. ApoA1 purification from blood plasma has always remained tedious, involving multiple steps, large volumes of plasma and substantial loss in the final yield of pure ApoA1. In this study, a two-step method has been developed and optimized for the purification of ApoA1 from plasma. Plasma was first subjected to 60% ammonium sulphate (NH ) SO precipitation and subsequently, ApoA1 was recovered using mixed mode chromatographic sorbent, HEA HyperCel™. ApoA1 was found to be enriched in 60% (NH ) SO supernatant that was dialyzed and injected onto HEA sorbent with 50 mM phosphate buffer pH 7.4. The bound proteins were eluted by decreasing the pH in step-gradient from pH 7.4 to pH 4.0 and subsequently to pH 3.5 using 50 mM sodium acetate buffer. Gel electrophoresis showed elution of homogeneous apoA1 at pH 3.5, with purity and yield of 63%. An interesting feature of this approach is that the purified ApoA1 was monomeric with a mass of 28,079.30 Da as confirmed by MS analysis. This simple and efficient method of purification of apoA1 serves as an alternative method which can be combined with traditional approaches and has a great potential for biochemical and clinical studies.
[Mh] Termos MeSH primário: Aminas/química
Apolipoproteína A-I/sangue
Apolipoproteína A-I/isolamento & purificação
Cromatografia Líquida/métodos
[Mh] Termos MeSH secundário: Sulfato de Amônio
Apolipoproteína A-I/química
Eletroforese em Gel de Poliacrilamida
Seres Humanos
Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (APOA1 protein, human); 0 (Amines); 0 (Apolipoprotein A-I); CI4E002ZV8 (hexylamine); SU46BAM238 (Ammonium Sulfate)
[Em] Mês de entrada:1803
[Cu] Atualização por classe:180302
[Lr] Data última revisão:
180302
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171218
[St] Status:MEDLINE


  6 / 125115 MEDLINE  
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[PMID]:28463693
[Au] Autor:Jeon EY; Choi BH; Jung D; Hwang BH; Cha HJ
[Ad] Endereço:Department of Chemical Engineering, Pohang University of Science and Technology, Pohang 790-784, South Korea.
[Ti] Título:Natural healing-inspired collagen-targeting surgical protein glue for accelerated scarless skin regeneration.
[So] Source:Biomaterials;134:154-165, 2017 Jul.
[Is] ISSN:1878-5905
[Cp] País de publicação:Netherlands
[La] Idioma:eng
[Ab] Resumo:Skin scarring after deep dermal injuries is a major clinical problem due to the current therapies limited to established scars with poor understanding of healing mechanisms. From investigation of aberrations within the extracellular matrix involved in pathophysiologic scarring, it was revealed that one of the main factors responsible for impaired healing is abnormal collagen reorganization. Here, inspired by the fundamental roles of decorin, a collagen-targeting proteoglycan, in collagen remodeling, we created a scar-preventive collagen-targeting glue consisting of a newly designed collagen-binding mussel adhesive protein and a specific glycosaminoglycan. The collagen-targeting glue specifically bound to type I collagen in a dose-dependent manner and regulated the rate and the degree of fibrillogenesis. In a rat skin excisional model, the collagen-targeting glue successfully accelerated initial wound regeneration as defined by effective reepithelialization, neovascularization, and rapid collagen synthesis. Moreover, the improved dermal collagen architecture was demonstrated by uniform size of collagen fibrils, their regular packing, and a restoration of healthy tissue component. Collectively, our natural healing-inspired collagen-targeting glue may be a promising therapeutic option for improving the healing rate with high-quality and effective scar inhibition.
[Mh] Termos MeSH primário: Colágeno/química
Hidrogel de Polietilenoglicol-Dimetacrilato/química
Adesivos Teciduais/química
Adesivos Teciduais/uso terapêutico
Cicatrização/efeitos dos fármacos
[Mh] Termos MeSH secundário: Animais
Colágeno Tipo I/química
Colágeno Tipo I/uso terapêutico
Decorina/química
Decorina/uso terapêutico
Eletroforese em Gel de Poliacrilamida
Feminino
Glicosaminoglicanos
Seres Humanos
Camundongos
Microscopia Eletrônica de Transmissão
Células NIH 3T3
Proteínas/química
Proteínas/uso terapêutico
Proteoglicanas/química
Proteoglicanas/uso terapêutico
Ratos
Ratos Sprague-Dawley
Pele/efeitos dos fármacos
Pele/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Collagen Type I); 0 (Decorin); 0 (Glycosaminoglycans); 0 (Proteins); 0 (Proteoglycans); 0 (Tissue Adhesives); 0 (adhesive protein, mussel); 25852-47-5 (Hydrogel, Polyethylene Glycol Dimethacrylate); 9007-34-5 (Collagen)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180228
[Lr] Data última revisão:
180228
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170503
[St] Status:MEDLINE


  7 / 125115 MEDLINE  
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[PMID]:29315346
[Au] Autor:Carvalho SB; Moreira AS; Gomes J; Carrondo MJT; Thornton DJ; Alves PM; Costa J; Peixoto C
[Ad] Endereço:iBET, Instituto de Biologia Experimental e Tecnológica, Oeiras, Portugal.
[Ti] Título:A detection and quantification label-free tool to speed up downstream processing of model mucins.
[So] Source:PLoS One;13(1):e0190974, 2018.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Mucins are high-molecular weight glycoproteins (0.25-20 MDa) containing one or more domains that are heavily O-glycosylated. Their implications as targets for cancer treatment have increased the interest in these glycoproteins, mainly in the fields of vaccines and antibodies. However, mucins present high heterogeneity, posing challenges that affect purification processes and quality control analysis. In that sense, it is necessary to develop and improve downstream processes and analytical methods to characterize these products. Here a tool based on biolayer interferometry analysis to improve mucin's detection and quantification in a fast, simple and label free-way is presented. Taking advantage of lectin recognition of mucins' carbohydrate structures, several lectins were evaluated and immobilized on streptavidin biosensors. Different assay conditions were optimized and the most suitable lectin, Aleuria aurantia lectin (AAL), was selected. Bovine Submaxillary Gland and human MUC5B mucins were used as proof of concept and were successfully detected and quantified at different stages of purification. High sensitivity levels were achieved with LOD and LOQ of 3.8 µg mL-1 and 11.7 µg mL-1 for BSM, and 0.2 µg mL-1 and 0.6 µg mL-1 for MUC5B. AAL binding specificity was also confirmed with fucose competition assays. Our method represents an advance on mucins detection and quantification since the existing methods present several disadvantages for process development. Hereafter, it can be applied to the optimization of new or already established downstream processes for mucins' purification.
[Mh] Termos MeSH primário: Modelos Moleculares
Mucinas/metabolismo
[Mh] Termos MeSH secundário: Cromatografia em Gel
Eletroforese em Gel de Poliacrilamida
Glicosilação
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (Mucins)
[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:180110
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0190974


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[PMID]:29324899
[Au] Autor:Kisidayová S; Pristas P; Zimovcáková M; Blanár Wencelová M; Homol'ová L; Mihaliková K; Cobanová K; Gresáková L; Váradyová Z
[Ad] Endereço:Institute of Animal Physiology, Slovak Academy of Sciences, Kosice, Slovakia.
[Ti] Título:The effects of high dose of two manganese supplements (organic and inorganic) on the rumen microbial ecosystem.
[So] Source:PLoS One;13(1):e0191158, 2018.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Little is known about the effects of the high dose and types of manganese supplements on rumen environment at manganese intake level close above the limit of 150 mg/kg of dry feed matter. The effects of high dose of two manganese supplements (organic and inorganic) on rumen microbial ecosystem after four months of treatment of 18 lambs divided into three treatment groups were studied. We examined the enzyme activities (α-amylase, xylanase, and carboxymethyl cellulase), total and differential microscopic counts of rumen ciliates, total microscopic counts of bacteria, and fingerprinting pattern of the eubacterial and ciliates population analyzed by PCR-DGGE. Lambs were fed a basal diet with a basal Mn content (34.3 mg/kg dry matter; control) and supplemented either with inorganic manganous sulfate or organic Mn-chelate hydrate (daily 182.7, 184 mg/kg dry matter of feed, respectively). Basal diet, offered twice daily, consisted of ground barley and hay (268 and 732 g/kg dry matter per animal and day). The rumens of the lambs harbored ciliates of the genera of Entodinium, Epidinium, Diplodinium, Eudiplodinium, Dasytricha, and Isotricha. No significant differences between treatment groups were observed in the total ciliate number, the number of ciliates at the genus level, as well as the total number of bacteria. Organic Mn did decrease the species richness and diversity of the eubacterial population examined by PCR-DGGE. No effects of type of Mn supplement on the enzyme activities were observed. In comparison to the control, α-amylase specific activities were decreased and carboxymethyl-cellulase specific activities were increased by the Mn supplements. Xylanase activities were not influenced. In conclusion, our results suggested that the intake of tested inorganic and organic manganese supplements in excess may affect the specific groups of eubacteria. More studies on intake of Mn supplements at a level close to the limit can reveal if the changes in microbial population impact remarkably the other rumen enzymatic activities.
[Mh] Termos MeSH primário: Ecossistema
Manganês/administração & dosagem
Microbiota
Rúmen/microbiologia
[Mh] Termos MeSH secundário: Ração Animal
Animais
Eletroforese em Gel de Poliacrilamida
Reação em Cadeia da Polimerase
Rúmen/metabolismo
Ovinos
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
42Z2K6ZL8P (Manganese)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180223
[Lr] Data última revisão:
180223
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:180112
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0191158


  9 / 125115 MEDLINE  
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[PMID]:29247504
[Au] Autor:Gallegos-Tabanico A; Sarabia-Sainz JA; Sarabia-Sainz HM; Carrillo Torres R; Guzman-Partida AM; Monfort GR; Silva-Campa E; Burgara-Estrella AJ; Angulo-Molina A; Acosta-Elias M; Pedroza-Montero M; Vazquez-Moreno L
[Ad] Endereço:Departamento de Física, Universidad de Sonora, Hermosillo, Sonora, 83000, México.
[Ti] Título:Molecular recognition of glyconanoparticles by RCA and E. coli K88 - designing transports for targeted therapy.
[So] Source:Acta Biochim Pol;64(4):671-677, 2017.
[Is] ISSN:1734-154X
[Cp] País de publicação:Poland
[La] Idioma:eng
[Ab] Resumo:The targeted drug delivery has been studied as one of the main methods in medicine to ensure successful treatments of diseases. Pharmaceutical sciences are using micro or nano carriers to obtain a controlled delivery of drugs, able to selectively interact with pathogens, cells or tissues. In this work, we modified bovine serum albumin (BSA) with lactose, obtaining a neoglycan (BSA-Lac). Subsequently, we synthesized glyconanoparticles (NPBSA-Lac) with the premise that it would be recognized by microbial galactose specific lectins. NPBSA-Lac were tested for bio-recognition with adhesins of E. coli K88 and Ricinus communis agglutinin I (RCA). Glycation of BSA with lactose was analyzed by electrophoresis, infrared spectroscopy and fluorescence. Approximately 41 lactoses per BSA molecule were estimated. Nanoparticles were obtained using water in oil emulsion method and spheroid morphology with a range size of 300-500 nm was observed. Specific recognition of NPBSA-Lac by RCA and E. coli K88 was displayed by aggregation of nanoparticles analyzed by dynamic light scattering and atomic force microscopy. The results indicate that the lactosylated nanovectors could be targeted at the E. coli K88 adhesin and potentially could be used as a transporter for an antibacterial drug.
[Mh] Termos MeSH primário: Antígenos de Bactérias/metabolismo
Portadores de Fármacos/metabolismo
Proteínas de Escherichia coli/metabolismo
Proteínas de Fímbrias/metabolismo
Nanopartículas/química
Lectinas de Plantas/metabolismo
[Mh] Termos MeSH secundário: Portadores de Fármacos/química
Eletroforese em Gel de Poliacrilamida
Escherichia coli/metabolismo
Lactose/química
Microscopia de Força Atômica
Peso Molecular
Tamanho da Partícula
Soroalbumina Bovina/química
Espectrofotometria Infravermelho
Espectroscopia de Infravermelho com Transformada de Fourier
Triptofano/química
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Antigens, Bacterial); 0 (Drug Carriers); 0 (Escherichia coli Proteins); 0 (K88 antigen, E coli); 0 (Plant Lectins); 0 (Ricinus communis agglutinin-1); 147680-16-8 (Fimbriae Proteins); 27432CM55Q (Serum Albumin, Bovine); 8DUH1N11BX (Tryptophan); J2B2A4N98G (Lactose)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180223
[Lr] Data última revisão:
180223
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171217
[St] Status:MEDLINE
[do] DOI:10.18388/abp.2017_1639


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[PMID]:29202487
[Au] Autor:Sarabia-Sainz HM; Mata Haro V; Sarabia Sainz JA; Vázquez-Moreno L; Montfort GR
[Ad] Endereço:Laboratorio de Bioquímica de Proteínas y Glicanos Coordinación de Ciencia de los Alimentos, Centro de Investigación en Alimentación y Desarrollo A.C., Hermosillo, Sonora 83304, México.
[Ti] Título:Maillard neoglycans as inhibitors for in vitro adhesion of F4 enterotoxigenic Escherichia coli to piglet intestinal cells.
[So] Source:Acta Biochim Pol;64(4):679-686, 2017.
[Is] ISSN:1734-154X
[Cp] País de publicação:Poland
[La] Idioma:eng
[Ab] Resumo:Adhesion of enterotoxigenic (ETEC) E. coli to host intestinal cells is mediated by lectin-like fimbriae that bind to specific glycan moieties on the surfaces of enterocytes. To prevent in vitro binding of E. coli F4 fimbriae (F4 ETEC ) to piglet enterocytes, neoglycans were synthesized by the Maillard reaction conjugating lactose (Lac), galacto-oligosaccharides (GOS) or chitin oligosaccharides (Ochit) to porcine serum albumin (PSA). Neoglycans were characterized by SDS-PAGE, intrinsic tryptophan fluorescence and recognition by plant lectins, as well as by F4 ETEC variants. Electrophoretic patterns suggested the binding to PSA of 63, 13 and 2 molecules of Lac, GOS and Ochit, respectively. All neoglycans displayed quenching of tryptophan fluorescence consistent with the degree of glycation estimated by SDS-PAGE. Plant lectins recognized the neoglycans according to their specificity, whereas antigenic variants of F4 ETEC (ab, ac and ad) recognized PSA-Ochit and PSA-Lac with higher affinity than that for GOS. Neoglycans partially hindered the in vitro binding of F4 ETEC to piglet enterocytes in a dose-dependent manner. The most effective blocking was observed with PSA-Lac that partially inhibited the adhesion of bacteria to enterocytes in a dose dependent manner, as quantified by flow cytometry. Increased production of the cytokines IL-6 and TNF-α was observed in response to F4 ETEC infection of enterocytes and production was reduced in the presence of PSA-Ochit and PSA-GOS. These results suggest that neoglycans synthesized by the Maillard reaction could be useful in the prophylaxis of diarrhea in piglets.
[Mh] Termos MeSH primário: Enterócitos/efeitos dos fármacos
Enterócitos/microbiologia
Escherichia coli Enterotoxigênica/efeitos dos fármacos
Polissacarídeos/química
Polissacarídeos/farmacologia
[Mh] Termos MeSH secundário: Animais
Antibacterianos/química
Antibacterianos/farmacologia
Aderência Bacteriana/efeitos dos fármacos
Eletroforese em Gel de Poliacrilamida
Escherichia coli Enterotoxigênica/patogenicidade
Infecções por Escherichia coli/tratamento farmacológico
Infecções por Escherichia coli/microbiologia
Infecções por Escherichia coli/veterinária
Produtos Finais de Glicação Avançada/química
Intestinos/citologia
Intestinos/microbiologia
Suínos
Doenças dos Suínos/virologia
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Anti-Bacterial Agents); 0 (Glycation End Products, Advanced); 0 (Polysaccharides)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180223
[Lr] Data última revisão:
180223
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171205
[St] Status:MEDLINE
[do] DOI:10.18388/abp.2017_2199



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