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Artigos de revistas sobre o tema "Isolation methods"

Autor: Grafiati

Publicado: 4 de junho de 2021

Última modificação: 20 de fevereiro de 2023

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1

Moťková, P., e J. Vytřasová. "Comparison of methods for isolating fungal DNA". Czech Journal of Food Sciences 29, Special Issue (4 de janeiro de 2012): S76—S85. http://dx.doi.org/10.17221/266/2011-cjfs.

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In this study methods of fungal DNA isolation were optimised and compared. The aim of the isolation processes was to obtain DNA of sufficient quality and quantity necessary for its amplification, as most detection techniques require DNA amplification before the proper DNA detection itself. For this purpose, classic methods of DNA extraction were compared and optimised while isolations using commercial kits were also done. The methods were evaluated from several perspectives, with focus especially laid on the isolated DNA not contain PCR inhibitors which would prevent DNA amplification, thus inhibiting the detection itself. For optimising the individual methods, collection strains of the genus Aspergillus were used. After the evaluation, two most suitable methods were selected and chosen for isolating potentially aflatoxigenic moulds taken from food samples. These methods were the commercially supplied kit for isolating DNA from plant leaves from Sigma and a classic method according Cenis in combination with the cell wall disruption by means of liquid nitrogen.

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2

Pariset, Eloise, Vincent Agache e Arnaud Millet. "Extracellular Vesicles: Isolation Methods". Advanced Biosystems 1, n.º 5 (24 de abril de 2017): 1700040. http://dx.doi.org/10.1002/adbi.201700040.

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3

Gilmore, Paul, e Umesh Gandhi. "Development of disc spring stack containment methods for vibration isolation". INTER-NOISE and NOISE-CON Congress and Conference Proceedings 263, n.º 2 (1 de agosto de 2021): 4871–79. http://dx.doi.org/10.3397/in-2021-2865.

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Cone disc springs exhibit quasi-zero stiffness behavior that is useful in isolating objects from low frequency vibrations. However, the stroke of a single disc spring is too low for most applications, and springs are stacked to increase the displacement. A method to contain the isolator stack then becomes critical for practical uses. Many challenges in developing these containment methods have been identified and can be collectively described as how to appropriately contain the stack without affecting isolation performance. In this work, three designs are considered: a retaining ring design, tube and shaft design, and zero poisson ratio sleeve design. Disc spring stacks with containment method are built, and load-deflection curves are measured and compared with standalone stacks. Under quasi-static compression testing, each containment method has minimal effect on the standalone stack load-deflection curve. However, significant differences in isolation performance are observed in vibration testing and found to depend on characteristics such as lateral stability, lateral strength, and degrees of freedom. Lastly, advantages, disadvantages, and appropriate applications for each containment method are summarized. The conclusions of this work are that containment method is an important variable in the application of disc spring isolators and robust, versatile containment designs have been demonstrated.

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4

Kallergi, Galatea, Eleni Politaki, Saad Alkahtani, Christos Stournaras e Vassilis Georgoulias. "Evaluation of Isolation Methods for Circulating Tumor Cells (CTCs)". Cellular Physiology and Biochemistry 40, n.º 3-4 (2016): 411–19. http://dx.doi.org/10.1159/000452556.

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Background: Detection of CTCs is a poor prognostic factor for many cancer types; however, their very low frequency represents an obstacle for their detection. The objective of the current study was to compare the performance of commonly used methods for CTCs isolation. Methods: The evaluated methods using spiking experiments of MCF7, SKBR3 and MDA MB-231 breast cancer cell lines were (i) ficoll density gradient separation (DGS), (ii) red blood cell lysis (Erythrolysis) isolation, (iii) positive immunomagnetic selection (EpCAM Dynal beads), (iv) two different negative immunomagnetic separation systems (Dynal vs Miltenyi CD45 beads) as well as (v) the Cell Search platform and (vi) the ISET system. Results: The recovery rates of Erythrolysis and DGS were 39% and 24%, respectively. Magnetic isolations are ranked from the worse to the best recovery rate as follows:, Myltenyi-anti-CD45 microbeads (24%); Dynal-anti-EpCAM beads (75%); Dynabeads-anti-CD45 (97%). CTCs isolation from blood samples using the CellSearch and ISET systems revealed that the recovery rate for Cell Search and ISET was 52% and 95%, respectively. Conclusions: Dynal-anti-CD45 beads have the best recovery rate compared to other magnetic methods. Furthermore the recovery rate of ISET was higher compared to Cell Search, especially for the more aggressive MDA-MB 231 cell line.

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5

Hosek, J., P. Svastova, M. Moravkova, I. Pavlik e M. Bartos. "Methods of mycobacterial DNA isolation from different biological material: a review". Veterinární Medicína 51, No. 5 (20 de março de 2012): 180–92. http://dx.doi.org/10.17221/5538-vetmed.

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Mycobacteria cause serious infections in animals and human beings. Huge economic losses on farms are caused by selected species of this wide family. A high risk of transmission of infection from animal to human exists. The knowledge of exact pathogen characteristics is an important factor which can improve quick and adequate healing. Cultivation and determination of phenotype is still the “gold standard”, but has the disadvantage of taking a long time and also low detection limit. Biochemical characterisation of isolates is not exact, and it is expensive. A more popular method used is the amplification of specific loci by polymerase chain reaction (PCR). For this method, the isolation of sufficient amounts of purified DNA is necessary. In this paper the most frequently used method for DNA isolation from live mycobacterial cells, body fluids, tissues, histological samples and forensic materials are outlined. This paper assists only as guide for these methods, so we describe them briefly.

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6

Sadrtdinova, G. R. "K. OXYTOCA BACTERIOPHAGES ISOLATION METHODS IMPROVEMENT". Russian Journal of Infection and Immunity 7, n.º 4 (1 de janeiro de 2017): 413–18. http://dx.doi.org/10.15789/2220-7619-2017-4-413-418.

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7

Fox, Gary L. "Methods and Apparatus for Isolation System". Journal of the Acoustical Society of America 130, n.º 2 (2011): 1083. http://dx.doi.org/10.1121/1.3625660.

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8

Veksler, Naum, Jean-Louis Izbicki, Jean-Marc Conoir e Pascal Rembert. "Methods of Isolation of Modal Resonances". Applied Mechanics Reviews 51, n.º 7 (1 de julho de 1998): 449–74. http://dx.doi.org/10.1115/1.3099015.

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The scattering problem by immersed targets involves the resonance phenomenon. Different methods of isolation of modal resonances are discussed: the Resonant Scattering Theory (the different backgrounds used in this method are considered), the phase gradient method (which is partly independent of the background choice), and the Argand diagram method (leading to both theoretical and experimental determination of the frequency and the width of resonances) and an exact description of the resonance components of partial modes (involving the determination of the roots of the characteristic equation in the complex frequency plane). The results provided by these methods are compared, their validity domain is discussed. Whatever the method, the resonant components appear as Breit-Wigner functions: this is the common point between the different methods. This review article includes 119 references.

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9

Teti, Anna, Anna Taranta, Ida Villanova, Irene Recchia e Silvia Migliaccio. "Osteoclast Isolation: New Developments and Methods". Journal of Bone and Mineral Research 14, n.º 7 (1 de julho de 1999): 1251–52. http://dx.doi.org/10.1359/jbmr.1999.14.7.1251.

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10

POWELL, T. "Methods of isolation of cardiac myocytes". Journal of Molecular and Cellular Cardiology 18 (1986): 30. http://dx.doi.org/10.1016/s0022-2828(86)80571-5.

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11

Barnes, Ella M. "Isolation methods for anaerobes in foods". International Journal of Food Microbiology 2, n.º 1-2 (junho de 1985): 81–87. http://dx.doi.org/10.1016/0168-1605(85)90060-1.

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12

Price, E. H., e G. H. Hunt. "Aeromonas in hospital—methods of isolation". Journal of Hospital Infection 8, n.º 3 (novembro de 1986): 309–11. http://dx.doi.org/10.1016/0195-6701(86)90131-3.

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13

Lopatin, S. A., G. N. Rudenskaya, I. M. Popova e T. M. Bikbov. "Chromatographic methods of mackerel proteases isolation". Food / Nahrung 35, n.º 5 (1991): 415–20. http://dx.doi.org/10.1002/food.19910350502.

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14

Holson, R. R., A. C. Scallet, S. F. Ali e B. B. Turner. "“Isolation stress” revisited: Isolation-rearing effects depend on animal care methods". Physiology & Behavior 49, n.º 6 (junho de 1991): 1107–18. http://dx.doi.org/10.1016/0031-9384(91)90338-o.

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15

Cronn, Richard, e Jonathan F. Wendel. "Simple methods for isolating homoeologous loci from allopolyploid genomes". Genome 41, n.º 6 (1 de dezembro de 1998): 756–62. http://dx.doi.org/10.1139/g98-078.

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During allopolyploid formation, divergent sets of chromosomes are combined in a common nucleus. Accordingly, loci that are single-copy in progenitor diploid genomes become duplicated, homoeologous loci in allopolyploids. Although homoeologous loci have been identified using classical genetic and linkage mapping approaches, there are few examples of the isolation and molecular characterization of homoeologous loci from allopolyploids. Here we describe two methods for the isolation of orthologous duplicated loci and demonstrate the feasibility of the techniques using allotetraploid cotton. The methods utilize restriction-digested, size-fractionated genomic DNA as a template for either plasmid cloning or PCR amplification. This fractionation procedure, when combined with Southern hybridization and mapping information, can separate homoeologues from each other and from more divergent cross-hybridizing sequences (paralogues). Each homoeologue can be then be isolated from a pool of size-fractionated genomic DNA that is enriched for target loci. While these methods were specifically designed for isolating homoeologous loci from allotetraploids, they should be applicable to a broad spectrum of diploid and polyploid plants and be useful for studying both ancient and recent gene duplications. In addition, the procedures enrich for orthologous sequences, which can be used for phylogenetic analysis. Thus, a general approach is detailed for the isolation of nuclear genes for phylogeny reconstruction.Key words: polyploidy, homoeology, orthology, Gossypium.

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16

Vaid, Rajesh Kumar, Taruna Anand, Priyanka Batra, Ram Avtar Legha e Bhupendra Nath Tripathi. "Evaluation of DNA Extraction Methods of Mule Dung". Defence Life Science Journal 4, n.º 3 (15 de julho de 2019): 170–74. http://dx.doi.org/10.14429/dlsj.4.14269.

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DNA isolation is a critical step in microbial community analysis of animal dung. DNA isolation from mule dung is challenging due to microbial diversity, composition and chemical nature of mule dung. Therefore, selection of an appropriate DNA isolation method is important to analyse the complete microbial diversity. In the current study, we evaluated the DNA isolation from mule dung samples (n=11) using QiAmp Mini stool kit as per manufacturer’s procedure with modifications. The results suggest that modifications in proprietary column based method improved the DNA quality and quantity suitable for mule dung microbial community analyses.

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17

Ruebsamen, Dale T. "Vibration Isolation Apparatus and Methods of Manufacture". Journal of the Acoustical Society of America 131, n.º 2 (2012): 1672. http://dx.doi.org/10.1121/1.3685657.

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18

Natonek-Wiśniewska, M., e E. Słota. "Methods of DNA isolation from poultry meal". Journal of Animal and Feed Sciences 16, n.º 3 (6 de setembro de 2007): 490–96. http://dx.doi.org/10.22358/jafs/66805/2007.

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19

Ponomarenko, E. A., O. V. Makarova, A. M. Kosyreva, M. V. Kondashevskaya, M. A. Diatroptova, I. S. Tsvetkov, A. A. Stepanov, L. P. Mikhailova e K. A. Artem’eva. "Comparative characteristics of spermatogenic cells isolation methods". CLINICAL AND EXPERIMENTAL MORPHOLOGY 8, n.º 3 (2019): 55–63. http://dx.doi.org/10.31088/cem2019.8.3.55-63.

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20

Chih-Yuan Lee, Tung-Sheng Chen e Chin-Hsing Kao. "Methods for noise isolation in RFCMOS ICs". IEEE Electron Device Letters 24, n.º 7 (julho de 2003): 478–80. http://dx.doi.org/10.1109/led.2003.815001.

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21

Bracho, Julio C., Arístides Rosado e Jorge A. Pino. "Comparison of Isolation Methods for Propolis Volatiles". Journal of Essential Oil Research 8, n.º 6 (novembro de 1996): 665–68. http://dx.doi.org/10.1080/10412905.1996.9701037.

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22

Jones, Leslie P. "Root isolation methods based upon lagrangian interpolation". International Journal of Computer Mathematics 24, n.º 3-4 (janeiro de 1988): 343–55. http://dx.doi.org/10.1080/00207168808803652.

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23

Schmidt, Stefan R., Fritz Schweikart e Martin E. Andersson. "Current methods for phosphoprotein isolation and enrichment". Journal of Chromatography B 849, n.º 1-2 (abril de 2007): 154–62. http://dx.doi.org/10.1016/j.jchromb.2006.09.016.

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24

HANKA, LADISLAV J., e RONDA D. SCHAADT. "Methods for isolation of streptoverticillia from soils." Journal of Antibiotics 41, n.º 4 (1988): 576–78. http://dx.doi.org/10.7164/antibiotics.41.576.

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25

Sloboda, Roger D. "Isolation of Microtubules by Assembly/Disassembly Methods". Cold Spring Harbor Protocols 2015, n.º 1 (janeiro de 2015): pdb.prot081182. http://dx.doi.org/10.1101/pdb.prot081182.

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26

Taylor, Harry Owen, Stephanie Herbers, Samuel Talisman e Nancy Morrow-Howell. "Assessing Social Isolation: Pilot Testing Different Methods". Journal of Gerontological Social Work 59, n.º 3 (2 de abril de 2016): 228–33. http://dx.doi.org/10.1080/01634372.2016.1197354.

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27

Lozhkin, A. V., e E. I. Sakanyan. "Natural coumarins: Methods of isolation and analysis". Pharmaceutical Chemistry Journal 40, n.º 6 (junho de 2006): 337–46. http://dx.doi.org/10.1007/s11094-006-0123-6.

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28

Williamson, Elizabeth M. "Methods in Biotechnology 4. Natural Products Isolation". Phytotherapy Research 13, n.º 5 (agosto de 1999): 451. http://dx.doi.org/10.1002/(sici)1099-1573(199908/09)13:5<451::aid-ptr531>3.0.co;2-p.

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29

Otsuru, S., T. Hofmann, P. Raman, T. Olson e E. Horwitz. "Equivalent MSC preparations using two isolation methods". Cytotherapy 16, n.º 4 (abril de 2014): S75. http://dx.doi.org/10.1016/j.jcyt.2014.01.276.

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30

Shvydkaya, Maria G., A. M. Zatevalov, S. D. Mitrokhin e D. T. Dzhandarova. "Comparison of culture and isolation methods for Clostridioides difficile and other anaerobes from stool samples in a routine microbiological laboratory practice". Clinical Microbiology and Antimicrobial Chemotherapy 23, n.º 2 (2021): 212–16. http://dx.doi.org/10.36488/cmac.2021.2.212-216.

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Objective. To improve stool sample culture and isolation of anaerobic flora, including Clostridioides difficile in the routine microbiological laboratory practice at the children’s oncology hospital. Materials and Methods. A total of 517 stool samples collected from patients in children’s oncology hospital from 2013 to 2015 were studied. All samples were analyzed by ELISA for C. difficile toxins and by culture according to dedicated 5 schemes for isolation of anaerobic bacteria, including C. difficile. Statistical significance of differences in isolation rates between the studied groups (culture schemes) was assessed by Pearson test. Results. Culture in liver broth and covering with technical agar followed by culture on anaerobic agar yielded 100% isolation rate of toxigenic C. difficile strains. This culture scheme is also suitable for isolating concomitant anaerobic flora: non-toxigenic C. difficile strains, Clostridium perfringens, other Clostridia spp. and Bacteroides spp. Conclusions. Use of the liquid accumulation medium and covering with technical agar make it possible to isolate anaerobic flora from stool samples and increase an isolation rate of toxigenic C. difficile strains to 100% of ELISA-positive samples.

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31

Ponomareva, N. I., A. P. Kostyusheva, S. A. Brezgin, V. V. Smirnov, V. I. Gegechkory, D. S. Kostyushev e V. P. Chulanov. "Comparison of exosome isolation methods for biomedical research". Journal Biomed 17, n.º 3E (26 de outubro de 2021): 76–79. http://dx.doi.org/10.33647/2713-0428-17-3e-76-79.

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Exosomes are nanoscale membrane vesicles secreted by almost all kinds of cells and stably existing in the body. Due to their unique properties, exosomes can be used in biomedical research as regenerative preparations or drug delivery vehicles. In this study, an anion exchange (AIEX) chromatography-based protocol for isolation of extracellular vesicles from a cell-conditioned medium was developed. The exosome isolation method based on AIEX chromatography overperforms the canonical ultracentrifugation-based isolation method in terms of its capability of large-volume processing.

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32

Et. al., Cici Jennifer Raj J,. "Influence at Mass of the Base Isolation System in Affecting the Higher Modes of Vibration". Turkish Journal of Computer and Mathematics Education (TURCOMAT) 12, n.º 2 (10 de abril de 2021): 1809–15. http://dx.doi.org/10.17762/turcomat.v12i2.1518.

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Base Isolation is a technology of mitigating the effects due to earthquakes with the aspect of dissipating the seismic waves away from the superstructure, by isolating the superstructure from the ground.This concept is widely essential to be implemented in structures(buildings) irrespective of many factors. There are several materials which could be implemented as base isolator, however the need in reduction of the number of the isolators is essential dueto various factors which a developing country finds difficult to implement. In this paper, a three-storey unsymmetrical building to be considered for the study is isolated by varying the mass of the foundation beam, (Transfer beam) thereby reducing the number of isolators in the building.Furthermore,the mode shapes and frequencies of the structure without base isolation and with base isolation considering mass of the base isolation system as a key factor were analysed and compared and hence the variation in the mass of the isolation system has a promising effect in altering the higher modes of vibration. The analysis is prolonged using another methd using UBC-1997 provisions and compared. In both the methods, the influence of the mass of the isolation system has a remarkable effect in altering the higher modes of vibration.

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Agarwal, Anuj, Ashish Kumar, Bhuvnesh Kumar Singh, Neelanchal Trivedi e K. K. Jha. "A review on extraction and phytochemical screening methods". Research in Pharmacy and Health Sciences 2, n.º 2 (15 de maio de 2016): 130–37. http://dx.doi.org/10.32463/rphs.2016.v02i02.25.

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Plants are the source of different drugs belonging to various therapeutic categories like antidiabetics, antispasmodics, antihypertensive, anticancer, antidepressants, antimicrobials, etc. Plants are used to treat various ailments and these plants have been used by different individuals and tribals worldwide. Use of plants to treat various ailments have also been mentioned in Ayurveda. Along these lines, various researchers are involved in isolating and assessing different bioactive molecules, to be isolated from various plant sources. Isolation of bioactive molecules is not an easy task for researchers. This review gives a focus on extraction and phytochemical screening methods along with their merits and demerits.

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Tereza, Sovová, Křížová Barbora e Ovesná Jaroslava. "Determining the optimal method for DNA isolation from fruit jams". Czech Journal of Food Sciences 36, No. 2 (7 de maio de 2018): 126–32. http://dx.doi.org/10.17221/340/2017-cjfs.

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DNA extraction is a crucial step in PCR analysis especially when analysing food samples that can be degraded and can potentially contain PCR-inhibiting substances. In this study, we compared the suitability of three DNA extraction methods – two kits: DNeasy<sup>®</sup> Plant Mini Kit and NucleoSpin<sup>®</sup> Food, and the CTAB method – for DNA extraction from commercial fruit jams. Fourteen jams with different contents of fruit, sugar and other additives were extracted in triplicate using the above-mentioned methods directly and after a washing step. The concentration and optical density were analysed using UV spectrophotometry and the amplifiability of the obtained DNA was evaluated using a PCR assay targeting a sequence coding for chloroplast tRNA-Leu. Samples isolated using the NucleoSpin<sup>®</sup> Food kit contained non-amplifiable DNA in eight cases, and samples isolated using the CTAB method could not be quantified. The DNeasy<sup>®</sup> Plant Mini Kit thus proved to be the most suitable method, since well-amplifiable DNA was obtained for all the analysed samples.

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Dijk, Manon van, t. holt, tte Severin, Suzanne Polinder e M. Vos. "The Daily Direct Costs of Isolating Patients Identified With Highly Resistant Microorganisms". Infection Control & Hospital Epidemiology 41, S1 (outubro de 2020): s404. http://dx.doi.org/10.1017/ice.2020.1053.

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Background: Isolation precautions are recommended when caring for patients identified with highly resistant microorganisms (HRMOs). However, the direct costs of isolating patients are largely unknown. Therefore, we aimed to obtain detailed information on the daily direct costs associated with isolating patients identified with HRMO. Methods: This study was performed from November until December 2017 on a 12-bed surgical ward. This ward contained solely isolation rooms with an anteroom. The daily direct costs of isolation were based on three cost items: (1) additional personal protective equipment (PPE); measured by counting the consumption of empty packaging materials, (2) cleaning and disinfection of the isolation room; based on the costs of an outsourced cleaning company, and (3) additional workload for healthcare workers; based on literature and multiplied by the average gross hourly salary of nurses. A distinction was made between the costs for strict isolation, contact-plus isolation, and contact isolation. Results: During the study period, 26 patients were nursed in isolation because of HRMO carriage, resulting in a total of 304 isolation days (median 7 isolation days; range 1-44). Gloves were consumed the most and hair caps the least. The average daily direct costs of isolation were the least expensive for contact isolation, €28/$31, and the most expensive for strict isolation, €41/$47. Conclusions: By using a novel, easy method to estimate consumption of PPE, we conclude that the daily direct costs of isolating a patient, differs per type of isolation. Insight into the direct costs of isolation is of utmost importance when developing or revising policies.Funding: NoneDisclosures: None

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Janowski, Daniel, Robin Wilgan, Tomasz Leski, Leszek Karliński e Maria Rudawska. "Effective Molecular Identification of Ectomycorrhizal Fungi: Revisiting DNA Isolation Methods". Forests 10, n.º 3 (1 de março de 2019): 218. http://dx.doi.org/10.3390/f10030218.

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A better understanding of ectomycorrhizal symbiosis leads to numerous advancements in forest management and environmental protection. The morphological identification of the ectomycorrhizae often proves to be misleading. For this reason, in order to study the ectomycorrhizal fungi communities, a number of molecular methods that require the isolation of nucleic acids are being used. However, ectomycorrhizal root tips, low mass heterogenic material rich in inhibitors, are a recalcitrant substrate in DNA isolation. It is common for published studies to include some number of unidentified root tips in their results, in spite of diverse isolation protocols being available to researchers. This study aims to analyze the relationship between the collected fungal material and later isolation results, and to propose a DNA isolation protocol specifically optimized for ectomycorrhizal root tips. It was found that the taxonomic position can be used to predict the potential isolation efficiency, with Ascomycota being generally more difficult from which to isolate DNA. After a number of cell lysis and lysate purification methods were evaluated, the joined approach of mechanical and chemical lysis, followed by silica column purification, was found to provide the best results, even with recalcitrant material.

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Mercadal, Marina, Carolina Herrero, Olga López-Rodrigo, Manel Castells, Alexandre de la Fuente, Francesc Vigués, Lluís Bassas e Sara Larriba. "Impact of Extracellular Vesicle Isolation Methods on Downstream miRNA Analysis in Semen: A Comparative Study". International Journal of Molecular Sciences 21, n.º 17 (19 de agosto de 2020): 5949. http://dx.doi.org/10.3390/ijms21175949.

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Seminal plasma (SP) contains a unique concentration of miRNA, mostly contained in small extracellular vesicles (sEVs) such as exosomes, some of which could be clinically useful for diagnosis and/or prognosis of urogenital diseases such as prostate cancer (PCa). We optimized several exosome-EV isolation technologies for their use in semen, evaluating EV purifying effectiveness and impact on the downstream analysis of miRNAs against results from the standard ultracentrifugation (UC) method to implement the use of SP sEV_miRNAs as noninvasive biomarkers for PCa. Our results evidenced that commercial kits designed to isolate exosomes/EVs from blood or urine are mostly applicable to SP, but showed quantitative and qualitative variability between them. ExoGAG 3500× g and the miRCURY Cell/Urine/CSF 1500× g methods resulted as equivalent alternative procedures to UC for isolating exosomes/sEVs from semen for nanoparticle characteristics and quality of RNA contained in vesicles. Additionally, the expression profile of the altered semen sEV-miRNAs in PCa varies depending on the EV isolation method applied. This is possibly due to different extraction techniques yielding different proportions of sEV subtypes. This is evidence that the exosome-EV isolation method has a significant impact on the analysis of the miRNAs contained within, with important consequences for their use as clinical biomarkers. Therefore, miRNA analysis results for EVs cannot be directly extrapolated between different EV isolation methods until clear markers for delineation between microvesicles and exosomes are established. However, EV extraction methodology affects combined models (semen exosome miRNA signatures plus blood Prostate specific antigen (PSA) concentration for PCa diagnosis) less; specifically our previously described (miR-142-3p + miR-142-5p + miR-223-3p + PSA) model functions as molecular marker from EVs from any of the three isolation methods, potentially improving the efficiency of PSA PCa diagnosis.

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Merga, J. Y., A. J. H. Leatherbarrow, C. Winstanley, M. Bennett, C. A. Hart, W. G. Miller e N. J. Williams. "Comparison ofArcobacterIsolation Methods, and Diversity ofArcobacterspp. in Cheshire, United Kingdom". Applied and Environmental Microbiology 77, n.º 5 (30 de dezembro de 2010): 1646–50. http://dx.doi.org/10.1128/aem.01964-10.

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ABSTRACTThe aims of this study were, firstly, to compare five published methods for the isolation ofArcobacterspp. from animal feces in order to determine the most sensitive and specific method. Second, we analyzed the resulting isolates by multilocus sequence typing (MLST) in order to investigate the diversity of the isolates recovered. Third, we investigated the ability to recoverArcobacterspp. from frozen fecal samples. Seventy-seven fecal samples from cattle, sheep, and badgers were subjected to five isolation methods, based on published methods for the isolation ofArcobacterandCampylobacterspp. Thirty-nineArcobacter butzleriisolates were analyzed using a multilocus sequence typing scheme. The survival ofArcobacterspp. in frozen samples was investigated by freezing the fecal samples at −80°C for 7 days and then applying the same five isolation methods. The most sensitive and specific method used anArcobacter-specific broth in conjunction with modified charcoal cefoperazone deoxycholate agar (mCCDA) with added antibiotics. Freezing of fecal samples led to a reduction in the recovery ofArcobacterspp. by approximately 50%. The 39 allelic profiles obtained by MLST could be divided into 11 sequence types (STs). We have identified the most sensitive and specific method for the isolation ofArcobacterspp. from animal feces and demonstrated that the freezing of fecal samples prior to isolation reduces arcobacter recovery. MLST analysis of the isolates revealed a high level of diversity.

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Vernerova, Andrea, Luiz Felipe Garcia-Souza, Ondrej Soucek, Milan Kostal, Vit Rehacek, Lenka Kujovska Krcmova, Erich Gnaiger e Ondrej Sobotka. "Mitochondrial Respiration of Platelets: Comparison of Isolation Methods". Biomedicines 9, n.º 12 (8 de dezembro de 2021): 1859. http://dx.doi.org/10.3390/biomedicines9121859.

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Multiple non-aggregatory functions of human platelets (PLT) are widely acknowledged, yet their functional examination is limited mainly due to a lack of standardized isolation and analytic methods. Platelet apheresis (PA) is an established clinical method for PLT isolation aiming at the treatment of bleeding diathesis in severe thrombocytopenia. On the other hand, density gradient centrifugation (DC) is an isolation method applied in research for the analysis of the mitochondrial metabolic profile of oxidative phosphorylation (OXPHOS) in PLT obtained from small samples of human blood. We studied PLT obtained from 29 healthy donors by high-resolution respirometry for comparison of PA and DC isolates. ROUTINE respiration and electron transfer capacity of living PLT isolated by PA were significantly higher than in the DC group, whereas plasma membrane permeabilization resulted in a 57% decrease of succinate oxidation in PA compared to DC. These differences were eliminated after washing the PA platelets with phosphate buffer containing 10 mmol·L−1 ethylene glycol-bis (2-aminoethyl ether)-N,N,N′,N′-tetra-acetic acid, suggesting that several components, particularly Ca2+ and fuel substrates, were carried over into the respiratory assay from the serum in PA. A simple washing step was sufficient to enable functional mitochondrial analysis in subsamples obtained from PA. The combination of the standard clinical PA isolation procedure with PLT quality control and routine mitochondrial OXPHOS diagnostics meets an acute clinical demand in biomedical research of patients suffering from thrombocytopenia and metabolic diseases.

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Aziz, Mohammad A., Benedict Seo, Haizal M. Hussaini, Merilyn Hibma e Alison M. Rich. "Comparing Two Methods for the Isolation of Exosomes". Journal of Nucleic Acids 2022 (25 de outubro de 2022): 1–6. http://dx.doi.org/10.1155/2022/8648373.

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Exosomes are membrane-bound nanovesicles released by cells into their extracellular environment. They carry different types of RNA including mRNA which may be useful in the diagnosis of various diseases. Exosome isolation has been a challenge because of their small size; therefore, two exosome isolation methods were compared in this study. The Exoquick-TC PLUS™ exosome isolation kit (kit) was compared with the classic ultracentrifugation (UC) method for exosome isolation. In samples obtained using both methods, cryo-electron microscopy showed round or slightly elongated vesicles with diameters ranging from 50 to 150 nm and delimited by a bilayered membrane. Dynamic light scattering resulted in multiple peaks for kit exosomes, whereas a single peak was observed for UC exosomes. Significantly, more total RNA was present in UC exosomes in contrast to kit exosomes ( P < 0.0001 ). This was reflected in subsequent mRNA analysis using qPCR, where UC exosomes had lower Ct values compared to kit exosomes. In conclusion, exosome characterization revealed the presence of exosomes in both UC and the kit samples. The kit samples presented additional peaks from DLS which might be due to impurities. Overall, due to a higher total RNA and mRNA content, UC is a better option for subsequent mRNA analysis; nevertheless, the kit can still be used if an ultracentrifuge is not available as four out of the five genes selected were detected and quantified using the kit.

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Bonvin, Debora, Diego Chiappe, Marc Moniatte, Heinrich Hofmann e Marijana Mionić Ebersold. "Methods of protein corona isolation for magnetic nanoparticles". Analyst 142, n.º 20 (2017): 3805–15. http://dx.doi.org/10.1039/c7an00646b.

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Stam, Janine, Sabine Bartel, Rainer Bischoff e Justina C. Wolters. "Isolation of extracellular vesicles with combined enrichment methods". Journal of Chromatography B 1169 (abril de 2021): 122604. http://dx.doi.org/10.1016/j.jchromb.2021.122604.

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KOMATSU, Tsunehiko, e Toshiro NAGASAWA. "Isolation Methods of Megakaryocytes from Human Bone Marrow". Japanese Journal of Thrombosis and Hemostasis 4, n.º 1 (1993): 36–39. http://dx.doi.org/10.2491/jjsth.4.36.

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Paget, Michelle, Hilary Murray, Clifford J. Bailey e Richard Downing. "Human islet isolation: semi-automated and manual methods". Diabetes and Vascular Disease Research 4, n.º 1 (março de 2007): 7–12. http://dx.doi.org/10.3132/dvdr.2007.010.

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Shao-jun, Shou, Liu Jing-li e Guo Xin-sheng. "Methods for testing disturbance isolation of EO system". Journal of Applied Optics 36, n.º 4 (2015): 504–8. http://dx.doi.org/10.5768/jao201536.0401002.

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Feng, Jie, Yan Li e Yu Nie. "Methods of mouse cardiomyocyte isolation from postnatal heart". Journal of Molecular and Cellular Cardiology 168 (julho de 2022): 35–43. http://dx.doi.org/10.1016/j.yjmcc.2022.04.007.

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Dundar, Tolga Turan, Mustafa Aziz Hatiboglu, Zehragul Ergul, Mehmet Hakan Seyithanoglu, Elif Sozen, Saffet Tuzgen, Mehmet Yasar Kaynar e Erdal Karaoz. "Glioblastoma Stem Cells and Comparison of Isolation Methods". Journal of Clinical Medicine Research 11, n.º 6 (2019): 415–21. http://dx.doi.org/10.14740/jocmr3781.

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BOLTON, F. J. (Eric). "Campylobacter Infections: Food-borne Sources and Isolation Methods". Japanese Journal of Food Microbiology 24, n.º 4 (2007): 151–56. http://dx.doi.org/10.5803/jsfm.24.151.

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Veksler, Naum, Jean‐Mark Conoir, Jean‐Louis Izbicki e Pascal Rembert. "Methods of isolation of modal resonances (a review)." Journal of the Acoustical Society of America 99, n.º 4 (abril de 1996): 2599–603. http://dx.doi.org/10.1121/1.415302.

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Hayakawa, Masayuki. "Selective isolation methods and distribution of soil actinomycetes." Actinomycetologica 4, n.º 2 (1990): 103–12. http://dx.doi.org/10.3209/saj.4_103.

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