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[PMID]:28455252
[Au] Autor:Kwak HW; Shin M; Lee JY; Yun H; Song DW; Yang Y; Shin BS; Park YH; Lee KH
[Ad] Endereço:Research Institute of Agriculture and Life Sciences, Seoul National University, Seoul 151-921, Republic of Korea.
[Ti] Título:Fabrication of an ultrafine fish gelatin nanofibrous web from an aqueous solution by electrospinning.
[So] Source:Int J Biol Macromol;102:1092-1103, 2017 Sep.
[Is] ISSN:1879-0003
[Cp] País de publicação:Netherlands
[La] Idioma:eng
[Ab] Resumo:Electrospinning of aqueous gelatin solution obtained from bovine or porcine sources has been difficult to achieve without additional facilities, such as a temperature control oven or heating cover. Gelatin from cold-water fish has low contents of proline (Pro) and hydroxyproline (Hyp) compared with mammalian-derived gelatin. For this reason, the fish-derived gelatin maintains a sol state without showing gelation behavior at room temperature. In the present study, we prepared an ultrafine fish gelatin nanofibrous web by electrospinning from aqueous solutions without any additive polymers or temperature control facilities. The concentration and viscosity of fish gelatin are the most important factor in determining the electrospinnability and fiber diameter. Electrospinning of aqueous fish gelatin has the highest nanofiber productivity compared to other organic solvent systems. Using glutaraldehyde vapor (GTA), the water stability was improved and substantial enhancement was achieved in the mechanical properties. Finally, the cytotoxicity of a fish gelatin nanofibrous scaffold was evaluated based on a cell proliferation study by culturing human dermal fibroblasts (HDFs) compared with a fish gelatin film and nanofibrous mat from mammalian gelatin. The result shows better initial cell attachment and proliferation compared with the fish gelatin film and no significant difference compared with mammalian-derived gelatin nanofibrous mat. We expect that electrospinning of aqueous fish gelatin could be an effective alternative mammalian gelatin source.
[Mh] Termos MeSH primário: Eletricidade
Peixes
Gelatina/química
Nanofibras/química
Nanotecnologia
Água/química
[Mh] Termos MeSH secundário: Animais
Materiais Biocompatíveis/química
Materiais Biocompatíveis/farmacologia
Bovinos
Adesão Celular/efeitos dos fármacos
Proliferação Celular/efeitos dos fármacos
Fibroblastos/citologia
Fibroblastos/efeitos dos fármacos
Gelatina/farmacologia
Glutaral/química
Seres Humanos
Hidrólise
Reologia
Soluções
Viscosidade
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Biocompatible Materials); 0 (Solutions); 059QF0KO0R (Water); 9000-70-8 (Gelatin); T3C89M417N (Glutaral)
[Em] Mês de entrada:1803
[Cu] Atualização por classe:180309
[Lr] Data última revisão:
180309
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170430
[St] Status:MEDLINE


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[PMID]:29281685
[Au] Autor:Luebbert CC; Clarke TM; Pointet R; Frahm GE; Tam S; Lorbetskie B; Sauvé S; Johnston MJW
[Ad] Endereço:Centre for Biologics Evaluation, Biologics and Genetic Therapies Directorate, Health Canada, Ottawa, Ontario, Canada.
[Ti] Título:Nanoparticle size and production efficiency are affected by the presence of fatty acids during albumin nanoparticle fabrication.
[So] Source:PLoS One;12(12):e0189814, 2017.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:We have previously identified extensive glycation, bound fatty acids and increased quantities of protein aggregates in commercially available recombinant HSA (rHSA) expressed in Oryza sativa (Asian rice) (OsrHSA) when compared to rHSA from other expression systems. We propose these differences may alter some attributes of nanoparticles fabricated with OsrHSA, as studies have associated greater quantities of aggregates with increased nanoparticle diameters. To determine if this is the case, nanoparticles were fabricated with OsrHSA from various suppliers using ethanol desolvation and subsequent glutaraldehyde cross-linking. All nanoparticles fabricated with OsrHSA showed larger diameters of approximately 20 to 90nm than particles fabricated with either defatted bovine serum albumin (DF-BSA) (100.9 ± 2.8nm) or human plasma albumin (pHSA) (112.0 ± 4.0nm). It was hypothesized that the larger nanoparticle diameters were due to the presence of bound fatty acids and this was confirmed through defatting OsrHSA prior to particle fabrication which yielded particles with diameters similar to those fabricated with pHSA. For additional conformation, DF-BSA was incubated with dodecanoic acid prior to desolvation yielding particles with significantly larger diameters. Further studies showed the increased nanoparticle diameters were due to the bound fatty acids modulating electrostatic interactions between albumin nanoparticles during the desolvation and not changes in protein structure, stability or generation of additional albumin oligomers. Finally the presence of dodecanoic acid was shown to improve doxorubicin loading efficiency onto preformed albumin nanoparticles.
[Mh] Termos MeSH primário: Ácidos Graxos/química
Nanopartículas
Albumina Sérica/química
[Mh] Termos MeSH secundário: Cromatografia em Gel
Dicroísmo Circular
Etanol/química
Glutaral/química
Seres Humanos
Microscopia Eletrônica de Varredura
Tamanho da Partícula
Proteínas Recombinantes/química
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (Fatty Acids); 0 (Recombinant Proteins); 0 (Serum Albumin); 3K9958V90M (Ethanol); T3C89M417N (Glutaral)
[Em] Mês de entrada:1801
[Cu] Atualização por classe:180129
[Lr] Data última revisão:
180129
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171228
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0189814


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[PMID]:27770419
[Au] Autor:Milosavic NB; Prodanovic RM; Velickovic D; Dimitrijevic A
[Ad] Endereço:Division of Experimental Therapeutics, Department of Medicine, Columbia University, 630 West 168th Street, P&S 8-436, New York, NY, 10032, USA. nm2729@columbia.edu.
[Ti] Título:Macroporous Poly(GMA-co-EGDMA) for Enzyme Stabilization.
[So] Source:Methods Mol Biol;1504:139-147, 2017.
[Is] ISSN:1940-6029
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:One of the most used procedures for enzyme stabilization is immobilization. Although immobilization on solid supports has been pursued since the 1950s, there are no general rules for selecting the best support for a giving application. A macroporous copolymer of ethylene glycol dimethacrylate and glycidyl methacrylate (poly (GMA-co-EGDMA)) is a carrier consisting of macroporous beads for immobilizing enzymes of industrial potential for the production of fine chemicals and pharmaceuticals.
[Mh] Termos MeSH primário: Aspergillus niger/enzimologia
Reagentes para Ligações Cruzadas/química
Enzimas Imobilizadas/química
Etilenoglicóis/química
Glucana 1,4-alfa-Glucosidase/química
Glutaral/química
Metacrilatos/química
[Mh] Termos MeSH secundário: Aspergillus niger/química
Aspergillus niger/metabolismo
Estabilidade Enzimática
Enzimas Imobilizadas/metabolismo
Óxido de Etileno/química
Glucana 1,4-alfa-Glucosidase/metabolismo
Oxirredução
Porosidade
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Cross-Linking Reagents); 0 (Enzymes, Immobilized); 0 (Ethylene Glycols); 0 (Methacrylates); 0 (poly(glycidyl methacrylate-co-ethylene glycol dimethacrylate)); EC 3.2.1.3 (Glucan 1,4-alpha-Glucosidase); JJH7GNN18P (Ethylene Oxide); T3C89M417N (Glutaral)
[Em] Mês de entrada:1801
[Cu] Atualização por classe:180115
[Lr] Data última revisão:
180115
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:161023
[St] Status:MEDLINE


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[PMID]:27770417
[Au] Autor:Roy I; Mukherjee J; Gupta MN
[Ad] Endereço:Department of Biotechnology, National Institute of Pharmaceutical Education and Research (NIPER), Sector 67, S.A.S. Nagar, Punjab, India.
[Ti] Título:Cross-Linked Enzyme Aggregates for Applications in Aqueous and Nonaqueous Media.
[So] Source:Methods Mol Biol;1504:109-123, 2017.
[Is] ISSN:1940-6029
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Extensive cross-linking of a precipitate of a protein by a cross-linking reagent (glutaraldehyde has been most commonly used) creates an insoluble enzyme preparation called cross-linked enzyme aggregates (CLEAs). CLEAs show high stability and performance in conventional aqueous as well as nonaqueous media. These are also stable at fairly high temperatures. CLEAs with more than one kind of enzyme activity can be prepared, and such CLEAs are called combi-CLEAs or multipurpose CLEAs. Extent of cross-linking often influences their morphology, stability, activity, and enantioselectivity.
[Mh] Termos MeSH primário: Reagentes para Ligações Cruzadas/química
Enzimas Imobilizadas/química
Glutaral/química
[Mh] Termos MeSH secundário: Animais
Aspergillus niger/enzimologia
Burkholderia cepacia/enzimologia
Candida/enzimologia
Bovinos
Estabilidade Enzimática
Enzimas Imobilizadas/metabolismo
Hidrolases/química
Hidrolases/metabolismo
Lipase/química
Lipase/metabolismo
Penicilina Amidase/química
Penicilina Amidase/metabolismo
Poligalacturonase/química
Poligalacturonase/metabolismo
Agregados Proteicos
Soroalbumina Bovina/metabolismo
Temperatura Ambiente
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Cross-Linking Reagents); 0 (Enzymes, Immobilized); 0 (Protein Aggregates); 27432CM55Q (Serum Albumin, Bovine); EC 3.- (Hydrolases); EC 3.1.1.3 (Lipase); EC 3.2.1.15 (Polygalacturonase); EC 3.5.1.11 (Penicillin Amidase); T3C89M417N (Glutaral)
[Em] Mês de entrada:1801
[Cu] Atualização por classe:180115
[Lr] Data última revisão:
180115
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:161023
[St] Status:MEDLINE


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[PMID]:27770415
[Au] Autor:Knezevic-Jugovic ZD; Grbavcic SZ; Jovanovic JR; Stefanovic AB; Bezbradica DI; Mijin DZ; Antov MG
[Ad] Endereço:Department for Biochemical Engineering and Biotechnology, Faculty of Technology and Metallurgy, University of Belgrade, Karnegijeva 4, Belgrade, Serbia. zknez@tmf.bg.ac.rs.
[Ti] Título:Covalent Immobilization of Enzymes on Eupergit Supports: Effect of the Immobilization Protocol.
[So] Source:Methods Mol Biol;1504:75-91, 2017.
[Is] ISSN:1940-6029
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:A selection of best combination of adequate immobilization support and efficient immobilization method is still a key requirement for successful application of immobilized enzymes on an industrial level. Eupergit supports exhibit good mechanical and chemical properties and allow establishment of satisfactory hydrodynamic regime in enzyme reactors. This is advantageous for their wide application in enzyme immobilization after finding the most favorable immobilization method. Methods for enzyme immobilization that have been previously reported as efficient considering the obtained activity of immobilized enzyme are presented: direct binding to polymers via their epoxy groups, binding to polymers via a spacer made from ethylene diamine/glutaraldehyde, and coupling the periodate-oxidized sugar moieties of the enzymes to the polymer beads. The modification of the conventionally immobilized enzyme with ethylenediamine via the carbodiimide route seems to be a powerful tool to improve its stability and catalytic activity.
[Mh] Termos MeSH primário: Candida/enzimologia
Enzimas Imobilizadas/química
Compostos de Epóxi/química
Etilenodiaminas/química
Glutaral/química
Lipase/química
Polímeros/química
[Mh] Termos MeSH secundário: Acrilamidas/química
Aminação
Candida/química
Candida/metabolismo
Estabilidade Enzimática
Enzimas Imobilizadas/metabolismo
Lipase/metabolismo
Metacrilatos/química
Oxirredução
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Acrylamides); 0 (Enzymes, Immobilized); 0 (Epoxy Compounds); 0 (Ethylenediamines); 0 (Methacrylates); 0 (Polymers); 60V9STC53F (ethylenediamine); 93356-75-3 (Eupergit C); EC 3.1.1.3 (Lipase); K67NG89J77 (methacrylamide); T3C89M417N (Glutaral)
[Em] Mês de entrada:1801
[Cu] Atualização por classe:180115
[Lr] Data última revisão:
180115
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:161023
[St] Status:MEDLINE


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[PMID]:28823320
[Au] Autor:Yamamoto Y; Iino K; Shintani Y; Kato H; Kimura K; Watanabe G; Takemura H
[Ad] Endereço:Department of Thoracic, Cardiovascular and General Surgery, Kanazawa University, Kanazawa, Japan. Electronic address: yamamoto054@gmail.com.
[Ti] Título:Comparison of Aortic Annulus Dimension After Aortic Valve Neocuspidization With Valve Replacement and Normal Valve.
[So] Source:Semin Thorac Cardiovasc Surg;29(2):143-149, 2017 Summer.
[Is] ISSN:1532-9488
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Aortic valve replacement (AVR) remains the standard surgical intervention for aortic valve disease and is preferred by many surgeons, despite its associated clinical issues. The clinical efficacy of aortic valve neocuspidization (AVNeo) with glutaraldehyde-treated autologous pericardium, the Ozaki procedure, has recently been reported. Although it is presumed to preserve the normal aortic annulus motion, changes to the aortic annulus during the cardiac cycle after AVNeo remain unclear. From March to December 2014, aortic annular dimensions were measured for 23 patients; the sample included 8 patients who had undergone AVNeo, 10 patients with normal aortic valves, and 5 patients who had undergone AVR. Measurements were recorded using electrocardiography-gated multidetector computed tomography. Data were analyzed using automated aortic root analysis software. Postoperative peak pressure gradients for the AVNeo and AVR groups were compared. No statistically significant differences in annulus variation were observed between patients who had undergone AVNeo and those with normal aortic valves. Annular area was larger during systole than during diastole in both groups. Postoperative peak pressure gradients were significantly lower in the AVNeo group than in the AVR group. The results of the present study demonstrated that aortic annular dimensions after AVNeo are similar to the dimensions of normal aortic valves. This was evidenced using electrocardiography-gated multidetector computed tomography, previously reported as the most reliable method, to evaluate annulus motion during the cardiac cycle. Lower postoperative peak pressure gradients might underlie the observed changes. These advantages will help in rectifying AVR defects.
[Mh] Termos MeSH primário: Valva Aórtica/cirurgia
Bioprótese
Doenças das Valvas Cardíacas/cirurgia
Implante de Prótese de Valva Cardíaca/instrumentação
Próteses Valvulares Cardíacas
Pericárdio/transplante
[Mh] Termos MeSH secundário: Idoso
Idoso de 80 Anos ou mais
Valva Aórtica/diagnóstico por imagem
Valva Aórtica/fisiopatologia
Autoenxertos
Técnicas de Imagem de Sincronização Cardíaca
Estudos de Casos e Controles
Eletrocardiografia
Feminino
Fixadores
Glutaral
Doenças das Valvas Cardíacas/diagnóstico por imagem
Doenças das Valvas Cardíacas/fisiopatologia
Implante de Prótese de Valva Cardíaca/efeitos adversos
Implante de Prótese de Valva Cardíaca/métodos
Hemodinâmica
Seres Humanos
Masculino
Meia-Idade
Tomografia Computadorizada Multidetectores
Valor Preditivo dos Testes
Desenho de Prótese
Interpretação de Imagem Radiográfica Assistida por Computador
Recuperação de Função Fisiológica
Resultado do Tratamento
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Fixatives); T3C89M417N (Glutaral)
[Em] Mês de entrada:1709
[Cu] Atualização por classe:170912
[Lr] Data última revisão:
170912
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170822
[St] Status:MEDLINE


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[PMID]:28661652
[Au] Autor:Stecula A; Schlessinger A; Giacomini KM; Sali A
[Ad] Endereço:Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco , San Francisco, California 94158, United States.
[Ti] Título:Human Concentrative Nucleoside Transporter 3 (hCNT3, SLC28A3) Forms a Cyclic Homotrimer.
[So] Source:Biochemistry;56(27):3475-3483, 2017 Jul 11.
[Is] ISSN:1520-4995
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Many anticancer and antiviral drugs are purine or pyrimidine analogues, which use membrane transporters to cross cellular membranes. Concentrative nucleoside transporters (CNTs) mediate the salvage of nucleosides and the transport of therapeutic nucleoside analogues across plasma membranes by coupling the transport of ligands to the sodium gradient. Of the three members of the human CNT family, CNT3 has the broadest selectivity and the widest expression profile. However, the molecular mechanisms of the transporter, including how it interacts with and translocates structurally diverse nucleosides and nucleoside analogues, are unclear. Recently, the crystal structure of vcCNT showed that the prokaryotic homologue of CNT3 forms a homotrimer. In this study, we successfully expressed and purified the wild type human homologue, hCNT3, demonstrating the homotrimer by size exclusion profiles and glutaraldehyde cross-linking. Further, by creating a series of cysteine mutants at highly conserved positions guided by comparative structure models, we cross-linked hCNT3 protomers in a cell-based assay, thus showing the existence of hCNT3 homotrimers in human cells. The presence and absence of cross-links at specific locations along TM9 informs us of important structural differences between vcCNT and hCNT3. Comparative modeling of the trimerization domain and sequence coevolution analysis both indicate that oligomerization is critical to the stability and function of hCNT3. In particular, trimerization appears to shorten the translocation path for nucleosides across the plasma membrane and may allow modulation of the transport function via allostery.
[Mh] Termos MeSH primário: Proteínas de Membrana Transportadoras/química
Modelos Moleculares
[Mh] Termos MeSH secundário: Absorção Fisiológica
Animais
Linhagem Celular
Cromatografia em Gel
Simulação por Computador
Reagentes para Ligações Cruzadas/química
Glutaral/química
Seres Humanos
Bicamadas Lipídicas/química
Bicamadas Lipídicas/metabolismo
Proteínas de Membrana Transportadoras/genética
Proteínas de Membrana Transportadoras/metabolismo
Conformação Molecular
Peso Molecular
Mutagênese Sítio-Dirigida
Mutação
Domínios e Motivos de Interação entre Proteínas
Estrutura Quaternária de Proteína
Proteínas Recombinantes de Fusão/química
Proteínas Recombinantes de Fusão/metabolismo
Proteínas Recombinantes/química
Proteínas Recombinantes/metabolismo
Sus scrofa
Trítio
Uridina/metabolismo
[Pt] Tipo de publicação:COMPARATIVE STUDY; JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Cross-Linking Reagents); 0 (Lipid Bilayers); 0 (Membrane Transport Proteins); 0 (Recombinant Fusion Proteins); 0 (Recombinant Proteins); 0 (cif nucleoside transporter); 10028-17-8 (Tritium); T3C89M417N (Glutaral); WHI7HQ7H85 (Uridine)
[Em] Mês de entrada:1707
[Cu] Atualização por classe:170726
[Lr] Data última revisão:
170726
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170630
[St] Status:MEDLINE
[do] DOI:10.1021/acs.biochem.7b00339


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[PMID]:28577843
[Au] Autor:Martin BJ; Kaestner M; Peng M; Ross DB; Urschel S; West LJ; Rebeyka IM
[Ad] Endereço:Department of Surgery, University of Alberta, Edmonton, Canada. Electronic address: billiejean@ualberta.ca.
[Ti] Título:Glutaraldehyde Treatment of Allografts and Aortic Outcomes Post-Norwood: Challenging Surgical Decision.
[So] Source:Ann Thorac Surg;104(4):1395-1401, 2017 Oct.
[Is] ISSN:1552-6259
[Cp] País de publicação:Netherlands
[La] Idioma:eng
[Ab] Resumo:BACKGROUND: Glutaraldehyde (GA) treatment of allografts used for arch reconstruction prevents the immunologic sensitization that occurs with untreated allografts, but its use may cause tissue changes that predispose to recurrent obstruction. The objective was to determine whether GA treatment of allografts used in Norwood procedures increases the risk of recurrent aortic obstruction. METHODS: All infants who underwent a Norwood procedure between 2000 and 2015 were included. Cryopreserved pulmonary allografts were used for all arch reconstructions; starting in 2005 all were treated with GA before use. Complete follow-up was obtained, including survival, transplantation, and all repeat procedures. Competing risks analyses were used to assess for differences in aortic reintervention over time. RESULTS: Two hundred six infants (132 male) were included. There were 60 deaths and 14 transplantations; 5-year transplantation-free survival was 71.9%. GA treatment of patches (n = 142, 68.9%) was not predictive of death (hazard ratio [HR] 1.38, 95% confidence interval [CI]: 0.61 to 3.08). Fifty-five patients had at least one aortic reintervention and 31 patients (15.0%) required surgical aortic reintervention. At 1-year, freedom from all aortic reintervention was similar between patients with and without treated patches, but freedom from surgical aortic reintervention was lower in the treated group (87.6% versus 95.3%, p = 0.0256). GA treatment was not associated with the combined end point of catheter-based or surgical reintervention but was associated with specific need for surgical reintervention (HR 4.05, 95% CI: 1.19 to 13.77). CONCLUSIONS: GA treatment is associated with increased late surgical aortic reintervention. The advantages of decreased sensitization with GA treatment need to be balanced against the risk of aortic reobstruction.
[Mh] Termos MeSH primário: Aloenxertos/efeitos dos fármacos
Glutaral/farmacologia
Síndrome do Coração Esquerdo Hipoplásico/cirurgia
Procedimentos de Norwood/métodos
Artéria Pulmonar/transplante
Reoperação/estatística & dados numéricos
[Mh] Termos MeSH secundário: Feminino
Seguimentos
Seres Humanos
Recém-Nascido
Masculino
Procedimentos de Norwood/efeitos adversos
Artéria Pulmonar/efeitos dos fármacos
Risco
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
T3C89M417N (Glutaral)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171115
[Lr] Data última revisão:
171115
[Sb] Subgrupo de revista:AIM; IM
[Da] Data de entrada para processamento:170605
[St] Status:MEDLINE


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[PMID]:28571924
[Au] Autor:Naik SM; Anil AC
[Ad] Endereço:Academy of Scientific and Innovative Research (AcSIR), CSIR-National Institute of Oceanography (NIO), Dona Paula 403 004, Goa, India.
[Ti] Título:Long-term preservation of Tetraselmis indica (Chlorodendrophyceae, Chlorophyta) for flow cytometric analysis: Influence of fixative and storage temperature.
[So] Source:J Microbiol Methods;139:123-129, 2017 Aug.
[Is] ISSN:1872-8359
[Cp] País de publicação:Netherlands
[La] Idioma:eng
[Ab] Resumo:Immediate enumeration of phytoplankton is seldom possible. Therefore, fixation and subsequent storage are required for delayed analysis. This study investigated the influence of glutaraldehyde (GA) concentrations (0.25%, 0.5%, and 1%) and storage temperatures (-80°C , -80°C, -20°C, and 5°C) on Tetraselmis indica for flow cytometric analysis. Cell recovery, granularity, and membrane permeability were independent of GA concentration whereas cell size and chlorophyll autofluorescence were concentration dependent. After an initial cell loss (16-19%), no cell loss was observed when samples were stored at 5°C. Cell recovery was not influenced by storage temperature until 4months but later samples preserved at -80°C , -80°C, and -20°C resulted in ~41% cell loss. Although maximum cell recovery with minimal effect on cell integrity was obtained at 5°C, autofluorescence was retained better at -80°C and -80°C. This suggests that in addition to fixative, the choice of storage temperature is equally important. Thus for long-term preservation, especially to retain autofluorescence, the use of lower concentration (0.25%) of GA when stored at a lower temperature (-80°C and -80°C) while a higher concentration (1%) of GA when stored at a higher temperature (5°C) is recommended.
[Mh] Termos MeSH primário: Clorófitas
Temperatura Baixa
Fixadores/farmacologia
Citometria de Fluxo/métodos
Glutaral/farmacologia
Preservação Biológica/métodos
[Mh] Termos MeSH secundário: Sobrevivência Celular
Clorófitas/fisiologia
Fluorescência
Fatores de Tempo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Fixatives); T3C89M417N (Glutaral)
[Em] Mês de entrada:1711
[Cu] Atualização por classe:171107
[Lr] Data última revisão:
171107
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170603
[St] Status:MEDLINE


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[PMID]:28542539
[Au] Autor:Lyda TA; Wagner EL; Bourg AX; Peng C; Tomaraei GN; Dean D; Kennedy MS; Marcotte WR
[Ad] Endereço:Department of Genetics and Biochemistry, Clemson University, Clemson, South Carolina, United States of America.
[Ti] Título:A Leishmania secretion system for the expression of major ampullate spidroin mimics.
[So] Source:PLoS One;12(5):e0178201, 2017.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Spider major ampullate silk fibers have been shown to display a unique combination of relatively high fracture strength and toughness compared to other fibers and show potential for tissue engineering scaffolds. While it is not possible to mass produce native spider silks, the potential ability to produce fibers from recombinant spider silk fibers could allow for an increased innovation rate within tissue engineering and regenerative medicine. In this pilot study, we improved upon a prior fabrication route by both changing the expression host and additives to the fiber pulling precursor solution to improve the performance of fibers. The new expression host for producing spidroin protein mimics, protozoan parasite Leishmania tarentolae, has numerous advantages including a relatively low cost of culture, rapid growth rate and a tractable secretion pathway. Tensile testing of hand pulled fibers produced from these spidroin-like proteins demonstrated that additives could significantly modify the fiber's mechanical and/or antimicrobial properties. Cross-linking the proteins with glutaraldehyde before fiber pulling resulted in a relative increase in tensile strength and decrease in ductility. The addition of ampicillin into the spinning solution resulted in the fibers being able to inhibit bacterial growth.
[Mh] Termos MeSH primário: Materiais Biomiméticos
Fibroínas/biossíntese
Leishmania/metabolismo
[Mh] Termos MeSH secundário: Ampicilina/farmacologia
Antibacterianos/farmacologia
Materiais Biomiméticos/farmacologia
Reatores Biológicos
Western Blotting
Reagentes para Ligações Cruzadas/química
Escherichia coli
Fibroínas/química
Fibroínas/farmacologia
Fibroínas/ultraestrutura
Glutaral/química
Leishmania/genética
Indústria Manufatureira
Teste de Materiais
Microscopia Eletrônica de Varredura
Projetos Piloto
Proteínas Recombinantes/biossíntese
Proteínas Recombinantes/química
Proteínas Recombinantes/farmacologia
Proteínas Recombinantes/ultraestrutura
Soluções
Resistência à Tração
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Anti-Bacterial Agents); 0 (Cross-Linking Reagents); 0 (Recombinant Proteins); 0 (Solutions); 7C782967RD (Ampicillin); 9007-76-5 (Fibroins); T3C89M417N (Glutaral)
[Em] Mês de entrada:1709
[Cu] Atualização por classe:170918
[Lr] Data última revisão:
170918
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170526
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0178201



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