65 research outputs found

    Global Analysis of Circulating Immune Cells by Matrix-Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry

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    Background: MALDI-TOF mass spectrometry is currently used in microbiological diagnosis to characterize bacterial populations. Our aim was to determine whether this technique could be applied to intact eukaryotic cells, and in particular, to cells involved in the immune response. Methodology/Principal Findings: A comparison of frozen monocytes, T lymphocytes and polymorphonuclear leukocytes revealed specific peak profiles. We also found that twenty cell types had specific profiles, permitting the establishment of a cell database. The circulating immune cells, namely monocytes, T lymphocytes and polymorphonuclear cells, were distinct from tissue immune cells such as monocyte-derived macrophages and dendritic cells. In addition, MALDI-TOF mass spectrometry was valuable to easily identify the signatures of monocytes and T lymphocytes in peripheral mononuclear cells. Conclusions/Significance: This method was rapid and easy to perform, and unlike flow cytometry, it did not require any additional components such as specific antibodies. The MALDI-TOF mass spectrometry approach could be extended t

    Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry Identification of Mycobacteria in Routine Clinical Practice

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    Background: Non-tuberculous mycobacteria recovered from respiratory tract specimens are emerging confounder organisms for the laboratory diagnosis of tuberculosis worldwide. There is an urgent need for new techniques to rapidly identify mycobacteria isolated in clinical practice. Matrix-assisted laser desorption time-of-flight mass spectrometry (MALDI-TOF MS) has previously been proven to effectively identify mycobacteria grown in high-concentration inocula from collections. However, a thorough evaluation of its use in routine laboratory practice has not been performed. Methodology: We set up an original protocol for the MALDI-TOF MS identification of heat-inactivated mycobacteria after dissociation in Tween-20, mechanical breaking of the cell wall and protein extraction with formic acid and acetonitrile. By applying this protocol to as few as 10 5 colony-forming units of reference isolates of Mycobacterium tuberculosis, Mycobacterium avium, and 20 other Mycobacterium species, we obtained species-specific mass spectra for the creation of a local database. Using this database, our protocol enabled the identification by MALDI-TOF MS of 87 M. tuberculosis, 25M. avium and 12 non-tuberculosis clinical isolates with identification scores $2 within 2.5 hours. Conclusions: Our data indicate that MALDI-TOF MS can be used as a first-line method for the routine identification of heatinactivated mycobacteria. MALDI-TOF MS is an attractive method for implementation in clinical microbiology laboratories i

    «La relation de limitation et d’exception dans le français d’aujourd’hui : exceptĂ©, sauf et hormis comme pivots d’une relation algĂ©brique »

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    L’analyse des emplois prĂ©positionnels et des emplois conjonctifs d’ “exceptĂ©â€, de “sauf” et d’ “hormis” permet d’envisager les trois prĂ©positions/conjonctions comme le pivot d’un binĂŽme, comme la plaque tournante d’une structure bipolaire. PlacĂ©es au milieu du binĂŽme, ces prĂ©positions sont forcĂ©es par leur sĂ©mantisme originaire dĂ»ment mĂ©taphorisĂ© de jouer le rĂŽle de marqueurs d’inconsĂ©quence systĂ©matique entre l’élĂ©ment se trouvant Ă  leur gauche et celui qui se trouve Ă  leur droite. L’opposition qui surgit entre les deux Ă©lĂ©ments n’est donc pas une incompatibilitĂ© naturelle, intrinsĂšque, mais extrinsĂšque, induite. Dans la plupart des cas (emplois limitatifs), cette opposition prend la forme d’un rapport entre une « classe » et le « membre (soustrait) de la classe », ou bien entre un « tout » et une « partie » ; dans d’autres (emplois exceptifs), cette opposition se manifeste au contraire comme une attaque de front portĂ©e par un « tout » Ă  un autre « tout ». De plus, l’inconsĂ©quence induite mise en place par la prĂ©position/conjonction paraĂźt, en principe, tout Ă  fait insurmontable. Dans l’assertion « les Ă©cureuils vivent partout, sauf en Australie » (que l’on peut expliciter par « Les Ă©cureuils vivent partout, sauf [qu’ils ne vivent pas] en Australie »), la prĂ©position semble en effet capable d’impliquer le prĂ©dicat principal avec signe inverti, et de bĂątir sur une telle implication une sorte de sous Ă©noncĂ© qui, Ă  la rigueur, est totalement inconsĂ©quent avec celui qui le prĂ©cĂšde (si « les Ă©cureuils ne vivent pas en Australie », le fait qu’ils « vivent partout » est faux). NĂ©anmoins, l’analyse montre qu’alors que certaines de ces oppositions peuvent enfin ĂȘtre dĂ©passĂ©es, d’autres ne le peuvent pas. C’est, respectivement, le cas des relations limitatives et des relations exceptives. La relation limitative, impliquant le rapport « tout » - « partie », permet de rĂ©soudre le conflit dans les termes d’une somme algĂ©brique entre deux sous Ă©noncĂ©s pourvus de diffĂ©rent poids informatif et de signe contraire. Les valeurs numĂ©riques des termes de la somme Ă©tant dĂ©sĂ©quilibrĂ©es, le rĂ©sultat est toujours autre que zĂ©ro. La relation exceptive, au contraire, qui n’implique pas le rapport « tout » - « partie », n’est pas capable de rĂ©soudre le conflit entre deux sous Ă©noncĂ©s pourvus du mĂȘme poids informatif et en mĂȘme temps de signe contraire : les valeurs numĂ©riques des termes de la somme Ă©tant symĂ©triques et Ă©gales, le rĂ©sultat sera toujours Ă©quivalent Ă  zĂ©ro

    DOES SYSTEMATIC PROFESSIONAL DEVELOPMENT FOR SCIENCE TEACHERS OF ENGLISH LANGUAGE LEARNERS (ELLS) MEET THEIR PROFESSIONAL NEEDS AND WHAT IS THE RELATIONSHIP BETWEEN PERCEPTIONS OF PROFESSIONAL DEVELOPMENT AND SELF-EFFICACY TO TEACH SCIENCE TO ELLS?

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    ABSTRACTProfessional learning is essential for science teacher preparation to teach science and part of a national movement to prepare students, including English Language Learners, (ELLs) for the demands of a 21st century workforce. The purpose of this research was to explore a) what science teachers feel they need in professional development to meet the needs of teaching science to ELL students; b) if science teachers demonstrate best practices learned in PD when teaching ELLs; and c) if there is a relationship between perceptions of professional development and self-efficacy in teaching science to ELLs.. Little is known about what science teachers need in their professional learning to effectively teach science to ELLs. Four data collection phases were used to ascertain teacher needs, preparedness, self-efficacy, and their ability to teach science to ELLs. The data were collected through two surveys, interviews, classroom observations, and case studies. The data were analyzed using a correlation analysis of the two surveys or coding interviews and observations into themes in relation to each research question. Findings in this study revealed no correlation between perceptions of PD and self-efficacy when teaching science to ELLs. Professional learning needs for science teachers with 50%> ELLs included sustained PD in science and ESOL, language development strategies, technology and material resources, and support for teaching from additional personnel and administrators. Most teachers were utilizing best practices when teaching science to ELLs but to varying degrees and had high scores in self-efficacy though they believed they were not prepared to meet the needs of ELL’s. Consequently, discovering science teachers’ needs by engaging them, as stakeholders in a process to include their needs, will create a structure that can design PD which can promote science among ELLs and today’s cultural and diverse classrooms.Keywords: professional development, self-efficacy, ELLs

    The atypical subunit composition of respiratory complexes I and IV is associated with original extra structural domains in Euglena gracilis.

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    In mitochondrial oxidative phosphorylation, electron transfer from NADH or succinate to oxygen by a series of large protein complexes in the inner mitochondrial membrane (complexes I-IV) is coupled to the generation of an electrochemical proton gradient, the energy of which is utilized by complex V to generate ATP. In Euglena gracilis, a non-parasitic secondary green alga related to trypanosomes, these respiratory complexes totalize more than 40 Euglenozoa-specific subunits along with about 50 classical subunits described in other eukaryotes. In the present study the Euglena proton-pumping complexes I, III, and IV were purified from isolated mitochondria by a two-steps liquid chromatography approach. Their atypical subunit composition was further resolved and confirmed using a three-steps PAGE analysis coupled to mass spectrometry identification of peptides. The purified complexes were also observed by electron microscopy followed by single-particle analysis. Even if the overall structures of the three oxidases are similar to the structure of canonical enzymes (e.g. from mammals), additional atypical domains were observed in complexes I and IV: an extra domain located at the tip of the peripheral arm of complex I and a "helmet-like" domain on the top of the cytochrome c binding region in complex IV
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