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Philippe RIEGEL, Yann DUMONT, Jacques CROIZÉ, and Olivier DAUWALDER. "SYSTÈMES AUTOMATIQUES D’IDENTIFICATION BACTÉRIENNE." ACTUALITES PERMANENTES EN MICROBIOLOGIE CLINIQUE 18, no. 04 (December 1, 2019): 10. http://dx.doi.org/10.54695/apmc.18.04.1522.
Анотація:
Située au cœur des processus analytiques de bactériologieconventionnelle, l’identification bactérienne peut être réalisée àl’aide de méthodes phénotypiques, protéiques, immunologiqueset/ou moléculaires. Cependant, c’est à la fin des années 2000 quel’identification bactérienne en microbiologie clinique a connu une« révolution » avec l’introduction de la spectrométrie de massede type « Matrix Assisted Laser Desorption Ionization /Time ofFlight » [SM MALDI TOF] (Seng et al. 2009). Cette dernièrepermet l’obtention d’un résultat en quelques minutes, avec uninoculum très faible et avec des performances élevées (van Veen etal. 2010, Dubois et al. 2012, Martiny et al. 2012, Fang et al. 2012,Marko et al. 2012, and Vila et al. 2012). Néanmoins, l’achat et lamaintenance du spectromètre de masse et de la base de donnéesrestent couteux (Neville et al. 2011) et nécessitent un volume annueld’identifications supérieur à 12000/15000 identifications, seuil àpartir duquel le cout de l’identification bactérienne et fongiquedevient inférieur à ceux des galeries d’identification automatiséepar technique biochimique. D’autre part, la SM MALDI TOF nepermet pas de séparer des espèces phylogénétiquement prochesdont l’identification repose sur quelques caractères phénotypiques.Parmi ces espèces peut différentiables par la SM MALDI TOF onpeut citer entre autres Escherichia coli et Shigella sp, Klebsiellaspp, Raoultella spp, et certains streptocoques (se reporter auchapitre dédié sur la SM MALDI TOF). En deçà, l’identificationbactérienne automatisée par galerie biochimique conserve l’intérêtde pouvoir être réalisée sur les mêmes instruments que l’antibiogramme automatisé avec un niveau acceptable de performanceset un résultat en quelques heures. Ainsi, ce chapitre portera surl’étude de ces systèmes automatisés d’identification et exclue defacto, les identifications par spectrométrie de masse, par techniquesimmunologiques et par biologie moléculaire
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Padilla Jaramillo, Carlos A., Luis M. Díaz Sánchez, Marianny Y. Combariza Montañez, Cristian Blanco Tirado, and Aldo F. Combariza Montañez. "Photon Harvesting Molecules: Ionization Potential from Quantum Chemical Calculations of Phytoplanktonic Pigments for MALDI-MS Analysis." Orinoquia 25, no. 1 (June 16, 2021): 13–23. http://dx.doi.org/10.22579/20112629.676.
Анотація:
The Ionization Potential (IP) of chemical species is of paramount importance for the Matrix Assisted Laser Desorption/Ionization (MALDI) analyticaltechnique. Specifically, IPs are used in MALDI MS Electron Transfer (ET) as a parameter to select the matrix for a given family of chemical species. We useda quantum chemical methodology to computationally determine IPs for a set of photosensible phytoplanktonic pigments. These calculations could be used as a guide for MALDI matrix selection. IPs were determined using Koopman’s Theorem, via Geometry Optimization and Single Point Energy within the Restricted Closed-Shell Hartree-Fock (RHF) technique. Structures of a twenty-four set of pigments were geometrically optimized, and their IPsdetermined. Calculated IP’s are in close agreement to reported experimental IPs within an average 3.7% absolute error. Structural features of the chemical species studied have a closed relationship with their chemical properties and IP’s. Our results suggest that ET-MALDI matrices such as DCTB (IP = 8.5 eV) and CNPV-OCH3 (IP = 8.3 eV) could be more suitable to analyze these types of chemical species.
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Lequeux, Guillaume. "Bactériologie de mammites : quelle place pour le laboratoire d’analyses ?" Le Nouveau Praticien Vétérinaire élevages & santé 14 (November 2022): 80–89. http://dx.doi.org/10.1051/npvelsa/2023013.
Анотація:
Au laboratoire, les outils de diagnostic étiologique d’une infection mammaire chez les bovins reposent encore principalement sur la culture et l’identification bactérienne, d’autant plus avec l’apport important de la technologie Maldi-TOF pour l’identification bactérienne ces dernières années. L’identification par Maldi-TOF permet une détermination rapide, facile et fiable des espèces bactériennes isolées et ouvre également des perspectives en termes de caractérisation bactérienne. L’approche par PCR, disponible depuis une dizaine d’années, permet notamment d’améliorer la sensibilité de la détection, mais constitue une méthode plus sensible aux contaminations et d’interprétation parfois plus délicate que la culture bactérienne. Les outils de séquençage, plus facilement accessibles dorénavant, ouvrent également des perspectives intéressantes. La détermination de la sensibilité aux antibiotiques des pathogènes mammaires, bien que limitée dans ses indications, reste un outil indispensable au praticien dans sa prise de décision de thérapeutique antibiotique. La mise en œuvre de certaines de ces méthodes est envisageable en clinique vétérinaire, mais d’autres (Maldi-TOF, séquençage) resteront probablement réservées quasi-exclusivement au domaine du laboratoire d’analyses. Les approches PCR seront probablement amenées à pouvoir se déployer en ESV dans un avenir proche compte-tenu des nouvelles technologies à présent disponibles.
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Kernalléguen, A., F. Saint-Marcoux, S. El Bakhi, F. Vorspan, R. Zenobi, G. Léonetti, D. Lafitte, and A. L. Pélissier-Alicot. "Caractérisation de la cocaïne et de ses métabolites par imagerie MALDI-MS 2 et comparaison avec une technique MAMS-MALDI-MS 2 et LC-MS 2." Toxicologie Analytique et Clinique 30, no. 2 (June 2018): S37—S38. http://dx.doi.org/10.1016/j.toxac.2018.04.042.
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Ding, Feng, Yuna Qian, Zaian Deng, Jitai Zhang, Yongchao Zhou, Lan Yang, Fangyan Wang, Juping Wang, Zhihua Zhou, and Jianliang Shen. "Retraction: Size-selected silver nanoparticles for MALDI-TOF mass spectrometry of amyloid-beta peptides." Nanoscale 13, no. 16 (2021): 7862. http://dx.doi.org/10.1039/d1nr90081a.
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Pomastowski, Paweł, Justyna Walczak, Marta Gawin, Szymon Bocian, Wojciech Piekoszewski, and Bogusław Buszewski. "Correction: HPLC separation of casein components on a diol-bonded silica column with MALDI TOF/TOF MS identification." Analytical Methods 7, no. 16 (2015): 6916. http://dx.doi.org/10.1039/c5ay90054a.
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Dahraoui, Souhail, Soukaina El Abbassi, Sara Kaissi, Hafida Naoui, Laila Boumhil, Maryem Iken, Azeddine Ibrahimy, and Badre Eddine Lmimouni. "Spectrométrie de masse MALDI-TOF et biomarqueurs de l’infection aspergillaire." Journal de Mycologie Médicale 27, no. 3 (September 2017): e43. http://dx.doi.org/10.1016/j.mycmed.2017.04.099.
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Denis, J., M. Machouart, F. Morio, M. Sabou, C. Kaufmann-Lacroix, N. Contet-Audonneau, E. Candolfi, and V. Letscher-Bru. "Développement et validation du diagnostic d’espèce de Malassezia par MALDI-TOF." Journal de Mycologie Médicale 26, no. 2 (June 2016): e9. http://dx.doi.org/10.1016/j.mycmed.2016.04.023.
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Degand, N., and R. Ruimy. "Intérêts et les limites actuelles du MALDI-TOF en microbiologie clinique." Journal des Anti-infectieux 14, no. 4 (November 2012): 159–67. http://dx.doi.org/10.1016/j.antinf.2012.07.005.
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Eveillard, Marion, Even Rustad, Mikhail Roshal, Yanming Zhang, Amanda Ciardiello, Neha Korde, Malin Hultcrantz, et al. "Using MALDI-TOF mass spectrometry for tracking of minimal residual disease in peripheral blood from patients with multiple myeloma." Journal of Clinical Oncology 37, no. 15_suppl (May 20, 2019): e19525-e19525. http://dx.doi.org/10.1200/jco.2019.37.15_suppl.e19525.
Анотація:
e19525 Background: Minimal residual disease (MRD) negativity after completed therapy is associated with longer progression-free survival (PFS) in patients with multiple myeloma (MM). Current standard of care for MRD testing use flow cytometry and/or next generation sequencing (NGS)-based assays applied on bone marrow (BM) aspirate samples. To develop a strategy for MRD tracking in peripheral blood (PB), we were motivated to evaluate MALDI-TOF head-to-head with established bone marrow-based MRD assays. Methods: We used MALDI-TOF mass spectrometry to detect M-proteins in PB. Our cohort included patients who had serum samples available at 2 timepoints including during active disease and within 60 days of MRD results as determined by flow cytometry of BM aspirates. The cohort enrolled 71 patients (26 females, 45 males) with a median age of 61 years (37-78 years). Twenty-seven patients had high-risk cytogenetics at baseline. Patients were classified at diagnosis as ISS1 (n = 38), ISS2 (n = 18) or ISS3 (n = 6). The flow cytometry based MRD assay was performed using MSKCCs 10-color, single-tube method. MALDI-TOF analysis was performed as described by Mills et al. Samples taken during active disease were used to identify the mass/charge ratio of the M-protein at baseline and in follow-up samples. MALDI-TOF results were compared to flow cytometry bone marrow-based MRD results. Results: The median time between diagnosis and the MRD timepoint was 13.4 months (3.4-91 months). MALDI-TOF in PB and flow cytometry BM-based MRD results were concordant for 44/71 (62%) patients (8+/+, 36 -/- respectively) while 27 were discordant (10 +/-, 17-/+). Fifty-four of 71 patients were in complete response (CR) (45/54 in sCR) at the time of MRD. MALDI-TOF was still positive in 13 of these 54 CR patients. In this cohort, the median PFS since MRD assessment was not reached in the 2 subgroups of double negative patients (n = 31) or in patients with a positive result in at least one technique (n = 23) with a median follow-up of 11.2 months (0-34.6 months). Conclusions: In 44/71 (62%) samples, MALDI-TOF of PB results and flow cytometry BM-based MRD results were concordant. MALDI-TOF of PB may be useful for detecting measurable residual disease and for the monitoring of MM patients during maintenance therapy with the future goal to rule out early recurrent disease.
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Guichou, Erwan. "Le génotypage par spectrométrie de masse MALDI-TOF : principe et applications biologiques." Journal de Mycologie Médicale 25, no. 3 (September 2015): 218. http://dx.doi.org/10.1016/j.mycmed.2015.06.005.
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Enomoto, Hirofumi, Tomohiro Furukawa, Shiro Takeda, Hajime Hatta, and Nobuhiro Zaima. "Unique Distribution of Diacyl-, Alkylacyl-, and Alkenylacyl-Phosphatidylcholine Species Visualized in Pork Chop Tissues by Matrix-Assisted Laser Desorption/Ionization–Mass Spectrometry Imaging." Foods 9, no. 2 (February 16, 2020): 205. http://dx.doi.org/10.3390/foods9020205.
Анотація:
Phosphatidylcholine (PC) is the major phospholipid in meat and influences meat qualities, such as healthiness. PC is classified into three groups based on the bond at the sn-1 position: Diacyl, alkylacyl, and alkenylacyl. To investigate their composition and distribution in pork tissues, including longissimus thoracis et lumborum (loin) spinalis muscles, intermuscular fat, and transparent tissues, we performed matrix-assisted laser desorption/ionization–mass spectrometry imaging (MALDI–MSI). Eleven diacyl-, seven alkylacyl-, and six alkenylacyl-PCs were identified using liquid chromatography (LC)-tandem MS (MS/MS) analysis. Despite many alkylacyl- and alkenylacyl-PC species sharing identical m/z values, we were able to visualize these PC species using MALDI–MSI. Diacyl- and alkylacyl- and/or alkenylacyl-PC species showed unique distribution patterns in the tissues, suggesting that their distribution patterns were dependent on their fatty acid compositions. PCs are a major dietary source of choline in meat, and the amount was significantly higher in the muscle tissues. Consumption of choline mitigates age-related memory decline and neurodegenerative diseases; therefore, the consumption of pork muscle tissues could help to mitigate these diseases. These results support the use of MALDI–MSI analysis for assessing the association between PC species and the quality parameters of meat.
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Dekio, Itaru, Yuki Sugiura, Susumu Hamada-Tsutsumi, Yoshiyuki Murakami, Hiroto Tamura, and Makoto Suematsu. "What Do We See in Spectra?: Assignment of High-Intensity Peaks of Cutibacterium and Staphylococcus Spectra of MALDI-TOF Mass Spectrometry by Interspecies Comparative Proteogenomics." Microorganisms 9, no. 6 (June 8, 2021): 1243. http://dx.doi.org/10.3390/microorganisms9061243.
Анотація:
Matrix-assisted laser-desorption/ionization time-of-flight (MALDI–TOF) mass spectrometry is a widely used and reliable technology to identify microbial species and subspecies. The current methodology is based on spectral fingerprinting, analyzing protein peaks, most of which are yet to be characterized. In order to deepen the understanding of these peaks and to develop a more reasonable identification workflow, we applied proteogenomic approaches to assign the high-intensity peaks of MALDI–TOF spectra of two bacterial genera. First, the 3–22 kD proteomes of 5 Cutibacterium strains were profiled by UPLC–MS/MS, and the amino acid sequences were refined by referring to their genome in the public database. Then, the sequences were converted to m/z (x-axis) values based on their molecular masses. When the interspecies comparison of calculated m/z values was well-fitted to the observed peaks, the peak assignments for the five Cutibacterium species were confirmed. Second, the peak assignments for six Staphylococcus species were performed by using the above result for Cutibacterium and referring to ribosomal subunit proteins coded on the S10-spc-alpha operon (the S10-GERMS method), a previous proteomics report by Becher et al., and comprehensive genome analysis. We successfully assigned 13 out of 15 peaks for the Cutibacterium species and 11 out of 13 peaks for the Staphylococcus species. DNA-binding protein HU, the CsbD-like protein, and 50S ribosomal protein L7/L12 were observed in common. The commonality suggests they consist of high-intensity peaks in the MALDI spectra of other bacterial species. Our workflow may lead to the development of a more accurate species identification database of MALDI–TOF mass spectrometry based on genome data.
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Bosco, Gabriella. "Marco Nuti, Et in pictura fabulator. Paul Cézanne et le dialogue créateur entre peinture, littérature et philosophie de Balzac à Maldi." Studi Francesi, no. 157 (LIII | I) (May 1, 2009): 226–27. http://dx.doi.org/10.4000/studifrancesi.8429.
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Sitterle, E., J. P. Bouchara, B. Dauphin, J. L. Beretti, S. Giraud, A. Paugam, N. Hassouni, et al. "Utilisation du MALDI-TOF-MS pour l’identification rapide et precise des especes cliniques de Scedosporium et de Pseudallescheria." Journal de Mycologie Médicale 22, no. 1 (March 2012): 100–101. http://dx.doi.org/10.1016/j.mycmed.2011.12.012.
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McElvania TeKippe, Erin. "The Added Cost of Rapid Diagnostic Testing and Active Antimicrobial Stewardship: Is It Worth It?" Journal of Clinical Microbiology 55, no. 1 (November 9, 2016): 20–23. http://dx.doi.org/10.1128/jcm.02061-16.
Анотація:
ABSTRACT Rapid diagnostic testing reduces the turnaround time for pathogen identification in the clinical microbiology laboratory, but the impact on patient care and hospital costs is a matter of speculation. Patel et al. (J. Clin. Microbiol. 55:60–67, 2017, https://doi.org/10.1128/JCM.01452-16 ) investigate the impact of matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) in conjunction with active antimicrobial stewardship to determine if implementation is indeed worth the added costs.
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Gonçalves, R. F., C. R. Ferreira, C. M. B. Orlandi, V. C. Sartori, H. N. Ferreira, F. C. Gozzo, S. A. Saraiva, E. J. Pilau, and M. N. Eberlin. "111 SINGLE EQUINE EMBRYO LIPID FINGERPRINTING BY MASS SPECTROMETRY." Reproduction, Fertility and Development 23, no. 1 (2011): 160. http://dx.doi.org/10.1071/rdv23n1ab111.
Анотація:
Matrix-assisted laser desorption-ionization mass spectrometry (MALDI-MS) allows the lipid profile study of individual mammal embryos. The data collection is rapid, highly sensitive, can tolerate some level of impurities, and is easy to interpret. The aim of this study was to report the lipid profile obtained from a single equine embryo by MALDI-MS. Follicles ≥30 mm in diameter were monitored daily until ovulation (Day 0). The insemination was performed close to ovulation with fresh diluted semen, and the embryo recovery was performed on Day 9 (D9) post ovulation. The equine embryo was placed in 50/50 (v/v) methanol/phosphate buffer solution and transported at 4°C to the laboratory. MALDI-MS spectra were acquired in the positive ion mode using MALDI Synapt HDMS mass spectrometer (Waters, Manchester, UK) m/z 700–950 range. The sample was coated with a 1.2 μL matrix of 2,5-dihydroxybenzoic acid (DHB) 1.0 mL L–1 in methanol. Due to the equine embryo volume, it was possible to divide it and get two mass spectra, which were identical. Spectra processing was performed using the MassLynx 4.0 software (Waters, Manchester, UK). It was observed the presence protonated and sodiated species of sphingomyelins (SM), phosphatidylcholines (PC), and triacylglycerols (TAG). The most intense ions assigned by comparison with data obtained from bovine embryos were m/z 723.5 [PC (34:1) and loss of N(CH3)3]+, 725.5. [SM (16:0) + Na]+, 754.6 [PC (32:1) + Na]+, 778.6 [PC (36:1) + Na]+, 780.6 [PC (34:2) + Na]+ or [PC (36:5) + H]+, 782.6 [PC (36:4) + H]+ or [PC (34:1) + Na]+, 788.6 [PC (36:1) + H]+, 806.6 [PC (38:6) + H]+ or [PC (36:3) + Na]+, 808.6 [PC (38:5) + H]+ or [PC (36:2) + Na]+, 810.6 [PC (38:4) + H]+ or [PC (36:1) + Na]+, 907.7 [TAG (54:3) + Na]+ and 909.7 [TAG (54:2) + Na] +. Regarding the lipid profile by MALDI-TOF previously reported for oocytes and embryos of several species (Ferreira et al. 2010, J. Lipid Res., 51, 1218–1227), it detected similar lipid species, but with different relative intensities. Because of the single equine embryo volume and MALDI-MS technique sensitivity, we intend to observe if there will be differences between the lipid profile of the inner cell mass and trophoblast in the future. The analysis of a greater number of embryos as well as different development periods and MS/MS experiments will contribute to building a database of lipid profiles that allows a better understanding of the lipid profile physiology in equine embryos and the meaning of differences among other mammalian embryos. FAPESP (São Paulo Research Foundation).
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Tata, A., D. Zampieri, J. L. Gonçalves, V. G. Santos, P. A. C. Braga, C. R. Ferreira, D. M. Assis, et al. "164 RAPID IDENTIFICATION OF BACTERIA IN BOVINE SEMEN BY MATRIX-ASSISTED LASER DESORPTION/IONIZATION MASS SPECTROMETRY." Reproduction, Fertility and Development 25, no. 1 (2013): 230. http://dx.doi.org/10.1071/rdv25n1ab164.
Анотація:
Frozen bovine semen used in the IVF process can be a potential source of microorganisms that can prevent or disturb embryo development and cause issues with the sanitary certification for bovine embryo commercialization and export. Therefore, the aim of this work is to introduce a novel tool for the fast identification of the pathogens on the frozen semen based on the mass spectra of their ribosomal proteins analysed by matrix-assisted desorption/ionization-mass spectrometry (MALDI-MS). Thirty bovine semen samples, which were aliquots of commercial sealed straws used daily in the commercial IVF routine at In vitro Brasil Ltd. (Mogi Mirim, SP, Brazil), were used for this work. Fifty microlitres of semen were incubated in 10 mL of brain heart infusion broth (BHI) for 24 h at 37°C. If turbidity was observed, the bacterial cultures were submitted to bacterial extraction and mass spectrometric analysis according to Barreiro et al. (2010). The mass spectra were obtained using an AUTOFLEX MALDI TOF/TOF and were analysed with the database library MALDI Biotyper 3.0 software (Bruker Daltonik, Germany) at default settings. For each sample, the result was given by means of a log score with a maximum value of 3.0. In this study, only scores higher than 2.0 were considered, which provide confident species identification. The bacteria identified were Citrobacter freundii (2 samples), Stenotrophomonas maltophilia (4 samples), Enterobacter cloacae (6 samples) complex, Candida parapsilosis (2 samples), and Enterococcus mundtii (2 samples). Note that all the identified bacteria consistently match with the most common contaminants reported in literature for bovine frozen semen (Bielanskia et al. 2003). The capability of the technique to identify the bacteria without the ribosomal extraction (i.e. of bacteria pellets diluted in water and acetonitrile) was successful for the pellet of S. maltophilia, C. freundii, and E. cloacae complex with scores higher than 2.3, indicating a very high probability of the identification of the bacterial genus and the species. This can be explained by considering the capability of the mass spectrometric matrix to lyse the membrane of the bacteria and directly extract and then ionize the ribosomal proteins. In order to exclude the presence of a mixing of bacteria in the pellet, the colonies were properly isolated. The results matched with the ones obtained before the isolation. In order to confirm the MALDI-MS identification, the isolated bacteria from the bovine semen were also submitted to sequencing of region 16SrRNA. In conclusion, MALDI-MS technique was successfully applied for the identification of pathogens in the bovine semen. Experiments to evaluate the presence of microorganisms in media used for in vitro maturation, IVF, and in vitro culture of the bovine oocytes and embryos using this strategy are underway. This robust and fast approach is able to detect early contamination and allows prevention of economic losses and sanitary excellence in the bovine IVF process.
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Eveillard, Marion, Even H. Rustad, Mikhail Roshal, Yanming Zhang, Amanda Kathryn Ciardiello, Neha Korde, Malin Hultcrantz, et al. "Comparison of MALDI-TOF Mass Spectrometry Analysis of Peripheral Blood and Bone Marrow Based Flow Cytometry for Tracking Measurable Residual Disease (MRD) in Patients with Multiple Myeloma." Blood 134, Supplement_1 (November 13, 2019): 3060. http://dx.doi.org/10.1182/blood-2019-123838.
Анотація:
Introduction In multiple myeloma (MM), the absence of measurable residual disease (MRD) after completed therapy is associated with longer progression free survival. Different techniques are available to detect low levels of plasma cells in bone marrow (BM) either by flow cytometry or by next-generation sequencing as a gold standard of molecular methods. But these techniques are limited because they require a representative bone marrow sample obtained by an invasive procedure. Therefore, detecting low levels of disease in blood would be ideal, because serial sampling is much easier and fully representative, and it would allow for the detection of extramedullary disease. Mass spectrometry-based methods have been shown to be more sensitive for detecting monoclonal proteins (M-protein) in serum. In this study, we were motivated to evaluate MALDI-TOF mass spectrometry (MALDI-TOF MS) head-to-head with an established BM-based MRD assays. Patients and Methods This cohort included 71 patients treated at Memorial Sloan Kettering Cancer Center (MSKCC) who had serum samples available at 2 timepoints including during active disease and within 60 days of MRD results as determined by flow cytometry of bone marrow aspirates (Flow-BM-MRD). The cohort enrolled 26 females and 45 males with a median age of 61 years (range 37-78 years). Twenty-seven patients had high-risk cytogenetics at baseline. The median time between diagnosis and the MRD timepoint was 13.4 months (3.4-91 months). MALDI-TOF MS analysis was performed according to the method published by Mills et al. Immunoglobulins were purified from serum samples using CaptureSelect beads specific of each isotype and were then eluted from the beads. Light chains and heavy chains were separated by the addition of a reducing agent. Purified samples were mixed in matrix and spotted onto a stainless steel MALDI plate and were analyzed using a Microflex LT MALDI-TOF mass spectrometer (Bruker). Samples taken during active disease were used to identify the mass to charge ratio (m/z) of the M-protein and served as a surrogate marker in the analysis of subsequent samples. MALDI-TOF MS results were compared to the Flow-BM-MRD assay, performed using the MSKCC's ten-color, single-tube method. Results MALDI-TOF MS detected an M-protein in all 71 active disease samples and in 25 MRD samples. MALDI-TOF-MS results at the MRD timepoint were concordant with Flow-BM-MRD for 44/71 patients (p=0.342, chi-square test). Eight patients were positive and 36 negative by both techniques. Twenty-seven patients were discordant, including 10 patients detectable only by Flow-BM-MRD and 17 detectable only by MALDI-TOF MS. Among the 10 patients detectable by flow cytometry but not by MALDI, the median MRD level was 0.00092% (+<0.0001% - 0.011%). The M-protein could have been present but below the polyclonal background. Regarding the 17 patients positive only by MALDI-TOF-MS, the BM sample for flow analysis was not suitable for 3 patients due to hemodilution. The others 14 samples reached the target of sensitivity with a limit of detection of 0.0001%. Alternatively, the MALDI-TOF result could be a false positive in terms of disease detection. MS is likely not falsely detecting M-proteins and indeed, immunofixation was also positive in 11/17 of these samples. However, low levels of M-protein may not indicate the presence of active disease. Indeed, a confounding factor is that immunoglobulins have a long half-life in serum. To determine the clinical utility of more sensitive M-protein detection, we evaluated the clinical outcome for the 48 newly diagnosed MM patients in CR at the MRD timepoint with a median follow-up of 11 months. Of these 48 patients, 2 of the 3 that were positive by both techniques relapsed during follow-up. One out of 27 patients that were negative by both techniques relapsed. None of the 10 patients who were positive only by MALDI-TOF relapsed and 1 of the 8 patients who were positive only by Flow-BM-MRD relapsed. Conclusions This study is important because it is a first step in understanding how to use a more sensitive blood test for the follow-up of MM patients. MALDI-TOF MS analysis may provide complementary results to Flow-BM-MRD especially for the follow-up of patients in CR and during maintenance therapy to detect poor responders that would be positive by both techniques. In summary, our results suggest that MALDI-TOF may be quite useful for early detection of relapse. Disclosures Roshal: Physicians' Education Resource: Other: Provision of services; Celgene: Other: Provision of Services; Auron Therapeutics: Equity Ownership, Other: Provision of services. Hassoun:Celgene: Research Funding; Janssen: Research Funding; Novartis: Consultancy. Smith:Celgene: Consultancy, Patents & Royalties, Research Funding; Fate Therapeutics and Precision Biosciences: Consultancy. Lesokhin:Takeda: Consultancy, Honoraria; Serametrix Inc.: Patents & Royalties; Genentech: Research Funding; GenMab: Consultancy, Honoraria; BMS: Consultancy, Honoraria, Research Funding; Janssen: Research Funding; Juno: Consultancy, Honoraria. Mailankody:Juno: Research Funding; Celgene: Research Funding; Janssen: Research Funding; Takeda Oncology: Research Funding; CME activity by Physician Education Resource: Honoraria. Landgren:Abbvie: Membership on an entity's Board of Directors or advisory committees; Sanofi: Membership on an entity's Board of Directors or advisory committees; Karyopharm: Membership on an entity's Board of Directors or advisory committees; Amgen: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding; Celgene: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding; Theradex: Other: IDMC; Adaptive: Honoraria, Membership on an entity's Board of Directors or advisory committees; Merck: Other: IDMC; Janssen: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding; Takeda: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding.
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Dekio, Itaru, Andrew McDowell, Mitsuo Sakamoto, Shuta Tomida, and Moriya Ohkuma. "Proposal of new combination, Cutibacterium acnes subsp. elongatum comb. nov., and emended descriptions of the genus Cutibacterium, Cutibacterium acnes subsp. acnes and Cutibacterium acnes subsp. defendens." International Journal of Systematic and Evolutionary Microbiology 69, no. 4 (April 1, 2019): 1087–92. http://dx.doi.org/10.1099/ijsem.0.003274.
Анотація:
In 2016, division of the genus Propionibacterium into four distinct genera was proposed. As a consequence, the species Propionibacterium acnes was transferred to Cutibacterium gen. nov. as Cutibacterium acnes comb. nov. The three recently proposed subspecies of P. acnes were not, however, accommodated in this proposal. Following a very recent validation of a new combination for C. acnes subsp. defendens and an automatically created C. acnes subsp. acnes , we now propose the new combination, C. acnes subsp. elongatum comb. nov. The type strain of Cutibacterium acnes subsp. elongatum is JCM 18919T (=NCTC 13655T). On the basis of further genomic and phenotypic (haemolysis and MALDI-TOF mass spectrometry) analyses of these subspecies, we also provide emended descriptions of the genus Cutibacterium Scholz and Kilian 2016, C. acnes subsp. acnes (Gilchrist 1900) Nouioui et al. 2018, and C. acnes subsp. defendens (McDowell et al. 2016) Nouioui et al. 2018.
21
Ovchinnikova, N. A., A. E. Sinyakov, A. V. Streletskii, and N. B. Polyakov. "Composition and Structure of [WCl4{N(Et)C(O)}3Cl]: MALDI-TOF Mass Spectrometric Study." Russian Journal of Coordination Chemistry 30, no. 2 (February 2004): 84–86. http://dx.doi.org/10.1023/b:ruco.0000015078.08085.7e.
22
Tessier, Barbara, Jéremy Ficara, Oana Polotanu, and Odile Eloy. "Analyse de la technique de dépôt et de la valeur des scores pour les levures et filamenteux en identification MALDI-TOF." Journal de Mycologie Médicale 25, no. 3 (September 2015): 231. http://dx.doi.org/10.1016/j.mycmed.2015.06.034.
23
Huang, Chien-Hsun, Chih-Chieh Chen, Jong-Shian Liou, Ai-Yun Lee, Jochen Blom, Yu-Chun Lin, Lina Huang, and Koichi Watanabe. "Genome-based reclassification of Lactobacillus casei: emended classification and description of the species Lactobacillus zeae." International Journal of Systematic and Evolutionary Microbiology 70, no. 6 (June 1, 2020): 3755–62. http://dx.doi.org/10.1099/ijsem.0.003969.
Анотація:
Taxonomic relationships between Lactobacillus casei , Lactobacillus paracasei and Lactobacillus zeae have long been debated. Results of previous analyses have shown that overall genome relatedness indices (such as average nucleotide identity and core nucleotide identity) between the type strains L. casei ATCC 393T and L. zeae ATCC 15820T were 94.6 and 95.3 %, respectively, which are borderline for species definition. However, the digital DNA‒DNA hybridization value was 57.3 %, which was clearly lower than the species delineation threshold of 70 %, and hence raised the possibility that L. casei could be reclassified into two species. To re-evaluate the taxonomic relationship of these taxa, multilocus sequence analysis (MLSA) based on the concatenated five housekeeping gene (dnaJ, dnaK, mutL, pheS and yycH) sequences, phylogenomic and core genome multilocus sequence typing analyses, gene presence and absence profiles using pan-genome analysis, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) profiling analysis, cellular fatty acid compositions, and phenotype analysis were carried out. The results of phenotypic characterization, MLSA, whole-genome sequence-based analyses and MALDI-TOF MS profiling justified an independent species designation for the L. zeae strains, and supported an emended the description of the name of Lactobacillus zeae (ex Kuznetsov 1956) Dicks et al. 1996, with ATCC 15820T (=DSM 20178T=BCRC 17942T) as the type strain.
24
Sendid, B., P. Ducoroy, N. François, G. Lucchi, S. Spinali, O. Vagner, S. Damiens, A. Bonnin, D. Poulain, and F. Dalle. "Intérêt de la spectrométrie de masse MALDI-TOF pour l’identification des levures. Évaluation et utilisation en routine hospitalière à Dijon et à Lille." Bio tribune magazine 40, no. 1 (December 2011): 37–44. http://dx.doi.org/10.1007/s11834-011-0060-x.
25
Jedmowski, Christoph, Ahmed Ashoub, Tobias Beckhaus, Thomas Berberich, Michael Karas, and Wolfgang Brüggemann. "Comparative Analysis of Sorghum bicolor Proteome in Response to Drought Stress and following Recovery." International Journal of Proteomics 2014 (October 1, 2014): 1–10. http://dx.doi.org/10.1155/2014/395905.
Анотація:
The adaptive response of Sorghum bicolor landraces from Egypt to drought stress and following recovery was analyzed using two-dimensional difference gel electrophoresis, 2D-DIGE. Physiological measurements and proteome alterations of accession number 11434, drought tolerant, and accession number 11431, drought sensitive, were compared to their relative control values after drought stress and following recovery. Differentially expressed proteins were analysed by Matrix assisted laser desorption ionisation time-of-flight mass spectrometry, MALDI-TOF-MS. Alterations in protein contents related to the energy balance, metabolism (sensu Mewes et al. 1997), and chaperons were the most apparent features to elucidate the differences between the drought tolerant and sensitive accessions. Further alterations in the levels of proteins related to transcription and protein synthesis are discussed.
26
Eveque, M., P. Emans, B. Claes, F. Bouwman, R. M. A. Heeren, and B. Cillero-Pastor. "OP0240 A MULTIMODAL MASS SPECTROMETRY APPROACH REVEALS SPECIFIC CARTILAGE MOLECULAR PROFILES ASSOCIATED TO TYPE 2 DIABETIC PATIENTS." Annals of the Rheumatic Diseases 79, Suppl 1 (June 2020): 151.2–152. http://dx.doi.org/10.1136/annrheumdis-2020-eular.5399.
Анотація:
Background:Osteoarthritis (OA) is mainly characterized by the progressive deterioration of articular cartilage. Recent studies support that type 2 diabetes (TD2) is a risk factor to develop OA [1, 2]. However, the molecular cartilage profile of patients combining these two diseases remains unclear, and a better understanding of the different OA phenotypes should be considered for the development of personalized medicine.Matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI) is used to investigate the bimolecular distribution of proteins, lipids or metabolites through thein-situanalysis of tissue sections. Bottom-up proteomics focuses on the relative quantification of proteins. The combination of both technologies could be considered to reveal specific molecular profiles and help for patient classification.Objectives:The main goal of this study is to apply a multimodal mass spectrometry approach on cartilage to reveal specific lipidomic and proteomic profiles associated to TD2 patients.Methods:Human cartilages from OA (na=10) and OA/TD2 human patients (nb=10) were obtained from donors undergoing total knee joint replacement. Cartilage punches of 8*8mm were sectioned at 12 µm thickness for MALDI-MSI and bottom-up proteomics.For MALDI-MSI experiments (na=6; nb=6), norharmane matrix was sprayed over the samples for the detection of lipids. Experiments were then performed in positive ion polarity at 50 µm of lateral resolution using a RapifleX MALDI Tissue-typer instrument. LipostarMSI and in-house ChemomeTricks toolbox for MATLAB software were used for data processing and analysis.For bottom-up proteomics experiment (na=10; nb=10), proteins were extracted, separated using SDS-PAGE and digested prior to liquid chromatography separation coupled to an orbitrap MS Q-Exactive HF mass spectrometer. Proteome Discoverer, enrichR and Reactome software were used for data processing and analysis.Results:MALDI-MSI showed overall differences between OA and OA/TD2 patients based on their specific lipidomic profiles. In particular, sphingomyelin and phosphatidylcholine species were significantly more abundant in OA patients whereas lysolipids such as lysophosphatidylcholine species were mainly present in OA/TD2 patients, providing therefore phenotype-specific OA molecular panels. Additionally, we observed that phosphatidylcholine and sphingomyelin species were more present in the superficial layer of the cartilage whereas lysophosphatidylcholine species were more abundant in the deep layer (Fig. 1A, B).Proteomics experiments applied on cartilage enables the quantification of 114 proteins. Among those, 73 were overexpressed in OA samples whereas 41 were overexpressed in OA/TD2 patients. Among the differentially regulated proteins (Fig. 1C), phospholipase A2 was increased in the diabetic cohort, in line with the elevated level of lysolipids found in the imaging data. Our results also involved the fatty acid omega oxidation and the fatty acid biosynthesis pathways as relevant to explain this deregulation of the lipid metabolism.Conclusion:MALDI–MSI combined with proteomics experiments showed different profiles between OA and OA/TD2 patients and could be employed for patient classification.References:[1]Louati, K., et al., Association between diabetes mellitus and osteoarthritis: systematic literature review and meta-analysis. RMD Open, 2015.1(1): p. e000077.[2]Williams, M.F., et al.,Type 2 diabetes and osteoarthritis: a systematic review and meta-analysis.J Diabetes Complications, 2016.30(5): p. 944-50.Acknowledgments:The Dutch Province of Limburg and MUMC institutional grant.Disclosure of Interests:Maxime Eveque: None declared, Pieter Emans Shareholder of: Shareholder and cofounder start-up company Chondropeptix, Grant/research support from: Institution received grants from STW, ReumaNederland, InSciTE, Consultant of: Consultancy to Kiomed, Speakers bureau: Payment for lectures by Kiomed, Episurf, Britt Claes: None declared, Freek Bouwman: None declared, Ron M A Heeren: None declared, Berta Cillero-Pastor: None declared
27
Dong, Mengqi, Yufan Hu, Huijun Zhang, Xinyuan Lan, Xiaolu Ran, Yijia Li, Lu Gan, and Shuguang Han. "Ultrasound-assisted extraction of bayberry tannin and optimization using response surface methodology." BioResources 16, no. 1 (January 22, 2021): 1825–41. http://dx.doi.org/10.15376/biores.16.1.1825-1841.
Анотація:
The extraction of bayberry tannins has potential to maximize the utilization of a forest waste. This study employed a four-level central composite design through response surface methodology to optimize the extraction of tannin from bayberry barks through ultrasound-assisted extraction (UAE). The effects of solute to solvent ratio (STSR), solvent concentration (SC), extraction time (ET), and sonication temperature (ST) on the total extraction yield of total condensed tannin (TCT yield) and total phenolic content (TPC) were investigated. The extracts were characterized with matrix-assisted laser desorption-time of flight mass spectrometer (MALDI-TOF MS), nuclear magnetic resonance (NMR), and gel permeation chromatography (GPC). The optimized condition was reached when the STSR and ST were set at 1:57.16 g/mL and 71.3%, when the ET and the ST was 39.1 min and 48.75 °C. In these conditions, the TCT yield and TPC reached their maximum values of 17.55% and 365.01 mg GAE/g, respectively. Furthermore, the polyflavonoids of bayberry tannin ranged from dimers to heptamers, which were only composed of proanthocyanidins (PC) containing galloy groups.
28
Jamal, W., K. Iregbu, A. Fadhli, F. Khodakhast, P. Nwajiobi-Princewill, N. Medugu, and V. O. Rotimi. "A point-prevalence survey of carbapenem-resistant Enterobacteriaceae in two different cities in Kuwait and Nigeria." African Journal of Clinical and Experimental Microbiology 23, no. 4 (October 23, 2022): 358–68. http://dx.doi.org/10.4314/ajcem.v23i4.4.
Анотація:
Background: The family Enterobacteriaceae belongs to the order Enterobacterales, a large diverse group of Gramnegative, facultatively anaerobic bacteria that sometimes cause multidrug-resistant infections which treatment options are often challenging. They are the leading cause of nosocomial bloodstream infection (BSI) and urinary tract infections (UTI). The objective of the study was to carry out a point-prevalence survey of antimicrobial resistance and carbapenem-resistant Enterobacteriaceae (CRE) clinical isolates in two hospitals in Kuwait and Nigeria.Methodology: Clinically significant bacterial isolates of patients from Kuwait and Nigeria, identified by VITEK-2 and MALDI-TOF mass spectrometry analysis were studied. Susceptibility testing of selected antibiotics was performed using E-test and broth dilution methods. Genes encoding carbapenemase, β-lactamases, and extended-spectrum β-lactamases (ESBLs) were detected by conventional PCR and sequencing, and whole genome sequencing (WGS) analyses.Results: Of 400 isolates from Kuwait and Nigeria, 188 (47.0%) and 218 (54.5%) were Escherichia coli and 124 (31.0%) and 116 (29.0%) Klebsiella pneumoniae, respectively. The prevalence of CRE was 14.0% in Kuwait and 8.0% in Nigeria. The resistance rates of CRE isolates against colistin and tigecycline in Kuwait were 6.6% versus 25.0%, and in Nigeria were 14.2% versus 14.2%, respectively. blaOXA-181 gene was the commonest in CRE isolates in Kuwait and blaNDM-7 in Nigeria. The commonest ESBL gene among the CRE isolates was blaCTX-M-15 in both countries. AmpC resistance genes were present in only Kuwait isolates and mediated by blaEBC, blaCIT and blaDHA. WGS analysis of 12 selected CRE isolates with carbapenem MICs>32μg/ml but no detectable genes from conventional PCR, revealed the presence of multidrug efflux pump genes such as major facilitator superfamily antibiotic efflux pump and resistance-nodulation-cell division antibiotic efflux pump groups.Conclusion: The prevalence of CRE was higher among isolates from Kuwait than Nigeria and the genes encoding resistance in CRE were different. The presence of efflux pump was a main mechanism of resistance in most of the Nigerian CRE isolates.
Contexte: La famille des Entérobactéries appartient à l'ordre des Entérobactéries, un grand groupe diversifié de bactéries anaérobies facultatives à Gram négatif qui provoquent parfois des infections multirésistantes dont les options de traitement sont souvent difficiles. Ils sont la principale cause d'infections nosocomiales du sang (BSI) et d'infections des voies urinaires (UTI). L'objectif de l'étude était de mener une enquête sur la prévalence ponctuelle de la résistance aux antimicrobiens et des isolats cliniques d'entérobactéries résistantes aux carbapénèmes (CRE) dans deux hôpitaux au Koweït et au Nigeria.Méthodologie: Des isolats bactériens cliniquement significatifs de patients du Koweït et du Nigéria, identifiés par analyse par spectrométrie de masse VITEK-2 et MALDI-TOF, ont été étudiés. Les tests de sensibilité des antibiotiques sélectionnés ont été effectués à l'aide des méthodes de test E et de dilution en bouillon. Les gènes codant pour la carbapénémase, les β-lactamases et les β-lactamases à spectre étendu (BLSE) ont été détectés par PCR et séquençage conventionnels et analyses de séquençage du génome entier (WGS).Résultats: Sur 400 isolats du Koweït et du Nigéria, 188 (47,0%) et 218 (54,5%) étaient Escherichia coli et 124 (31,0%) et 116 (29,0%) Klebsiella pneumoniae, respectivement. La prévalence de la CRE était de 14,0% au Koweït et de 8,0% au Nigeria. Les taux de résistance des isolats CRE à la colistine et à la tigécycline au Koweït étaient de 6,6% contre 25,0%, et au Nigeria de 14,2% contre 14,2%, respectivement. Le gène blaOXA-181 était le plus courant dans les isolats CRE au Koweït et blaNDM-7 au Nigeria. Le gène BLSE le plus courant parmi les isolats CRE était blaCTX-M-15 dans les deux pays. Les gènes de résistance à l'AmpC étaient présents uniquement dans les isolats du Koweït et médiés par blaEBC, blaCIT et blaDHA. L'analyse WGS de 12 isolats CRE sélectionnés avec des CMI de carbapénème >32 μg/ml mais aucun gène détectable par PCR conventionnelle, a révélé la présence de gènes de pompe d'efflux multidrogues tels que la pompe d'efflux antibiotique de la superfamille facilitatrice majeure et les groupes de pompe d'efflux antibiotique de division cellulaire de résistance-nodulation.Conclusion: La prévalence de la CRE était plus élevée parmi les isolats du Koweït que du Nigeria et les gènes codant pour la résistance à la CRE étaient différents. La présence d'une pompe à efflux était un mécanisme principal de résistance dans la plupart des isolats CRE Nigérians.
29
Schumann, P., R. Pukall, C. Spröer, and E. Stackebrandt. "Reclassification of Koreibacter algae as a later heterotypic synonym of Paraoerskovia marina and emended descriptions of the genus Paraoerskovia Khan et al. 2009 and of Paraoerskovia marina Khan et al. 2009." International Journal of Systematic and Evolutionary Microbiology 63, Pt_1 (January 1, 2013): 219–23. http://dx.doi.org/10.1099/ijs.0.040600-0.
Анотація:
16S rRNA gene sequences deposited for the type strains of Paraoerskovia marina (CTT-37T; GenBank accession no. AB445007) and Koreibacter algae (DSW-2T; FM995611) show a similarity of 100 %. Consequently, the type strains were subjected to a polyphasic recharacterization under direct comparison in order to clarify their taxonomic position. PvuII RiboPrint patterns and quantitative ratios of cellular fatty acids revealed strain-specific differences between P. marina DSM 21750T ( = CTT-37T) and K. algae DSM 22126T ( = DSW-2T). The percentage of DNA–DNA binding of 94 % indicated that the two type strains belong to the same genomospecies. Agreement in the peptidoglycan structure and polar lipid pattern, highly similar fatty acid profiles and MALDI-TOF mass spectra, the ability to produce acid from the same carbon sources, corresponding enzymic activities and DNA G+C contents of 70.8±0.3 mol%, in addition to the consistent characteristics reported in the original descriptions, support the view that the two strains should be affiliated to the same species. According to Rules 38 and 42 of the Bacteriological Code, Koreibacter algae should be reclassified as later heterotypic synonym of Paraoerskovia marina , and the descriptions of the genus Paraoerskovia Khan et al. 2009 and of Paraoerskovia marina Khan et al. 2009 are emended accordingly.
30
Nakayama, Yuushou, Keiya Katagi, Ryo Tanaka, and Takeshi Shiono. "Ring-Opening Polymerization of Lactones Catalyzed by Silicon-Based Lewis Acid." International Journal of Polymer Science 2023 (May 17, 2023): 1–6. http://dx.doi.org/10.1155/2023/4391372.
Анотація:
Many catalysts containing various elements at their active sites have been reported for the ring-opening polymerization (ROP) of cyclic esters. However, to our knowledge, silicon-based catalysts for ROP have never been reported. Here we report the ROP of cyclic esters and cyclic carbonates catalyzed by the derivatives of bis(perchlorocatecholato)silane (Si(catCl)2), which is a neutral silicon-based Lewis acid recently reported by Greb et al. The catalyst systems show high activity for the ROP of seven- and six-membered ring monomers such as ε-caprolactone, δ-valerolactone, and trimethylene carbonate to produce the polymers with molecular weights up to 32 kg/mol. The matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and nuclear magnetic resonance analysis of the obtained polymers indicates the predominant formation of cyclic polymers.
31
Medimagh, Raouf, Saber Chatti, Sandra Alves, Noura Hammed, André Loupy, and Hédi Zarrouk. "Caractérisation par spectrométrie de masse MALDI–TOF de poly(isosorbide-éther)s cycliques et linéaires obtenus sous irradiation microondes." Comptes Rendus Chimie 10, no. 3 (March 2007): 234–50. http://dx.doi.org/10.1016/j.crci.2006.11.002.
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Kurachi, Sumiko, Taku Tanaka, Muneyoshi Kanai, Emi Suenaga, Elena Solovieva, and Kotoku Kurachi. "Age-Related Homeostasis: Molecular Mechanism, Disease and Predictions from Global Analyses of Mouse Liver Proteins." Blood 108, no. 11 (November 16, 2006): 1611. http://dx.doi.org/10.1182/blood.v108.11.1611.1611.
Анотація:
Abstract We previously reported the first molecular mechanism underlying the age-related homeostasis, ASE/AIE-mediated genetic mechanism for age-related regulation of gene expression, which turned out to be a mechanism of puberty-onset gene switching, specifically controlling a group of genes for their expression from puberty into old age (Kurachi & Kurachi J Thromb Haemost 2005; Zahng et al J Biol Chem277:4532, 2002; Kurachi et al Science285:739, 1999). This led us to successful construction of a transgenic mouse model of hemophilia B Leyden, a unique subset of hemophilia B with its mechanism being remained mysterious, robustly mimicking its unusual pattern of puberty-onset spontaneous amelioration. With this background, we hypothesized that besides the ASE/AIE-mediated mechanism, there exist more unidentified fundamental regulatory mechanisms for age-related homeostasis, which individually or in various combinations generate age-related complex and dynamic regulatory patterns of liver proteins. We launched a series of global and quantitative analysis of age-related changes in expression of liver nuclear proteins of mice (C57BL/6xSJL, [male], 1 through-24 month of age) by taking a procedure composed of two-dimensional gel electrophoresis (2DE) for separation and quantification of liver nuclear protein spots and of MALDI-TOF/MS PMF analyses to identify proteins in the spots. Out of over 6000 spots recognized and quantified in 2DE, 4547 protein spots were subjected to MALDI-TOF/MS analysis for protein identification. Finally, 2765 protein spots including many isomers were found unique. Systematic analyses of their age-related expression identified several major phases in protein expression throughout the lifespan. These findings supported our hypothesis that there exist multiple novel molecular mechanisms responsible for maintaining age-related homeostasis. The comprehensive liver nuclear protein data set was then used to construct a comprehensive database, which allows rapid and reliable searches for expression of specific proteins, their age-related dynamic profiles, isomers, protein identification from 2DE image, and other related information. This will serve as a valuable platform resource for studying epigenetic challenges, evaluation of drugs as well as gaining further insights into the molecular mechanisms underlying age-related homeostasis.
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Ravaonindrina, Noro, Iony Razanajatovo, and Alexandra Bastaraud. "Qualité microbiologique de la viande commercialisée dans la communauté urbaine d’Antananarivo." Revue d’élevage et de médecine vétérinaire des pays tropicaux 67, no. 3 (June 30, 2015): 122. http://dx.doi.org/10.19182/remvt.10178.
Анотація:
Certains entéropathogènes majeurs sont associés aux produits carnés, notamment Salmonella spp. ou Escherichia coli producteur de shigatoxines (3), et ce, en lien avec les conditions d’hygiène rencontrées de l’abattage à l’étal (1). L’objectif a été d’évaluer (a) le niveau des indicateurs d’hygiène comme E. coli, (b) la prévalence de Salmonella spp., de Campylobacter thermotolérants, de Listeria monocytogenes, d’Yersinia enterocolitica et d’E. coli O157:H7 dans les viandes commercialisées dans la communauté urbaine d’Antananarivo, et (c) la sensibilité aux antibiotiques des isolats.Au total 137 échantillons, incluant de la viande de zébu hachée (n = 52) ou non (n = 48), et de la viande de porc hachée (n = 10) ou non (n = 27) ont été collectés dans les grandes surfaces (24 p. 100) et sur les marchés d’Antananarivo (76 p. 100) d’avril à novembre 2014. Des méthodes conventionnelles ISO (International Organization for Standardization) et validées (Vidas technologie Biomérieux) ont été mises en oeuvre, ainsi que la confirmation des isolats par spectrométrie de masse (Maldi-TOFF, Bruker) ou par biologie moléculaire pour E. coli O157H7 (2). Le profil d’antibio-résistance des isolats a été déterminé sur Adagio Automated System (Biorad).Moins de 2 p. 100 des échantillons respectaient les critères microbiologiques d’hygiène du Règlement européen CE n° 2073/2005 (respectivement 70 et 90 p. 100 de non-conformité pour les micro-organismes aérobies et pour E. coli). Au total 111 échantillons (81 p. 100) étaient porteurs au moins d’un micro-organisme potentiellement pathogène ; 74 S. enterica subsp. enterica ont été isolées avec 33 sérovars identifiés dont deux prédominants, Budapest (21 p. 100) et Muenchen (21 p. 100) ; 48 E. coli O157:H7 ont été isolés principalement dans la filière bovine (63 p. 100) ; L. monocytogenes et Campylobacter thermotolérants ont été détectés respectivement dans 3 et 2 p. 100 des échantillons, et Y. enterocolitica n’a pas été détectée (tableau I).Les isolats de Salmonella ont été sensibles aux antibiotiques testés, hormis une S. enterica serovar Bahrenfeld qui a résisté aux fluoroquinolones, notamment à la ciprofloxacine. Par ailleurs, 35 souches d’E.coli O157:H7 testées présentaient un phénotype sauvage et huit isolats avaient un phénotype d’hyperproduction de céphalosporinases (tableau II).Les conditions d’hygiène rencontrées sur l’ensemble de la filière bovine et porcine sont nettement insuffisantes et contribuent à la propagation de pathogènes entériques, comme Salmonella enterica et le pathovar Escherichia coli O157:H7. Des phénotypes multirésistants aux antibiotiques émergent, représentant un facteur de risque pour la santé des consommateurs. Par conséquence, nous élargirons le champ des investigations aux abattoirs et aux élevages pour une caractérisation génotypique des isolats qui permettra de rechercher les sources et les modalités potentielles de contamination.
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Grigoroiu-Serbanescu, M., S. Herms, A. Georgi, J. Hauser, P. Czerski, C. C. Diaconu, G. Babadjanova, et al. "Bipolar and Unipolar Major Depression are not Associated with the P2RX7-gene (SNP rs2230912) in an European Case-control Sample." European Psychiatry 24, S1 (January 2009): 1. http://dx.doi.org/10.1016/s0924-9338(09)70451-1.
Анотація:
Background:In two recent studies the SNP rs2230912 (Gln460Arg) located in exon 13 of P2RX7-gene (chromosome 12q24) provided the strongest evidence of association with bipolar disorder (BP) (Barden et al,2006; McQuillin et al, 2008) and in one study with the unipolar major depression (Mdd-UP) ( Lucae et al, 2006).Objective:In the present study we investigated the involvement of the SNP rs2230912 in BPI in four European samples from Germany, Poland, Romania and Russia and in the combined sample (N=1445) in comparison with a combined sample of 2006 normal controls. Additionally, a Mdd-UP sample (N=640) from Germany was studied.Method:All patients were diagnosed according to DSM-IV-R. The BPI sample consisted of 802 females (55.5%) and 643 males (44.5%); the mean AO was 26.9 (SD=10.6); the mean age-at-interview was 42.7 (SD=12.5). The control sample had a mean age of 39.74 (SD=11.22) [1113 females (55.5%); 893 males (44.5%)]. Genotyping of all national samples was performed at Bonn University using the Mass ARRAY system on a Sequenom Compact MALDI-TOF-device. The single marker analysis was performed with FAMHAP software.Results:There was no allelic association between the G-allele of the SNP rs2230912 and either BPI or Mdd-UP both in the national samples and in the combined sample. The genotypic analysis also indicated no significant results. Our samples of patients and controls had genotypic distributions similar to those of the previously published studies and even in these studies there were no significant differences in genotype frequencies between BPI patients and controls.
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Moya-Lopez, Carmen, Ivan Bravo, José A. Castro-Osma, David Chapron, Patrice Bourson, Christelle Vagner, Marianne Cochez, et al. "Synthesis of High Molecular Weight Stereo-Di-Block Copolymers Driven by a Co-Initiator Free Catalyst." Polymers 14, no. 2 (January 7, 2022): 232. http://dx.doi.org/10.3390/polym14020232.
Анотація:
Stereo-diblock copolymers of high molecular weight polylactide (PLA) were synthetized by the one pot-sequential addition method assisted by a heteroscorpionate catalyst without the need of a co-initiator. The alkyl zinc organometallic heteroscorpionate derivative (Zn(Et)(κ3-bpzteH)] (bpzteH = 2,2-bis(3,5-dimethylpyrazol-1-yl)-1-para-tolylethoxide) proved to assist in the mechanism of reaction following a coordination-insertion process. Kinetic studies along with the linear correlation between monomer and number average molecular weight (Mn) conversion, and the narrow polydispersities supported the truly living polymerization character of the initiator, whereas matrix-assisted laser desorption/Ionization-time of flight (MALDI-TOF) studies showed a very low order of transesterification. The high stereo-control attained for the afforded high molecular weight derivatives was revealed by homonuclear decoupled 1H NMR spectra and polarimetry measurements. The nanostructure of the PLA derivatives was studied by both wide-angle X-ray scattering (WAXS) and differential scanning calorimetry (DSC) and the stereocomplex phase of the PLA stereo-diblock copolymers was successfully identified.
36
Qin, C. G., Y. Li, W. N. Niu, Y. Ding, X. Y. Shang, and C. L. Xu. "Composition analysis and structural identification of anthocyanins in fruit of waxberry." Czech Journal of Food Sciences 29, No. 2 (March 25, 2011): 171–80. http://dx.doi.org/10.17221/177/2010-cjfs.
Анотація:
Anthocyanin pigments in the fruit of waxberry (Myrica rubra Sieb. et Zucc.), were extracted with 0.1% HCl in ethanol, and the crude anthocyanin extract was purified by C18 Sep-Pak cartridge open-column chromatography. High-performance liquid chromatography (HPLC) with photodiode array detection (PAD) and matrix-assisted laser desorption/ionisation-time of flight-mass spectrometry (MALDI-TOF-MS) was applied for the separation and identification of anthocyanins in the fruit of waxberry and their aglycones resulting from acid hydrolysis. Three anthocyanins were found in the fruit of waxberry and identified as Cyanidin 3-O-β-galacopyranoside (14.8%), Cyanidin 3-O-β-gluco-<br />pyranoside (60.5%), and petunidin 3-O-β-glucopyranoside (24.7%), respectively, using spectroscopic methods (UV-Vis<br />and MS). The three anthocyanins were isolated and purified by preparative HPLC, and their chemical structures were further characterised by H<sup>1</sup> NMR. On the basis of chromatographic data, the total anthocyanin content was 286 mg/g in fresh fruit of waxberry.
37
BARBULESCU, Iuliana Diana, Razvan Ionut TEODORESCU, Corina DUMITRACHE, Mihaela BEGEA, Camelia Filoftea DIGUTA, Mihai FRINCU, Daniel Cornel BANITA, et al. "OBTAINING ACTIVE DRY YEASTS BIOMASS FOR THE PRODUCTION OF PIETROASA WINES." AgroLife Scientific Journal 12, no. 2 (December 31, 2023): 18–24. http://dx.doi.org/10.17930/agl202323.
Анотація:
Consumers are increasingly demanding traditional beverages such as wines that are obtained through innovative technologies and that use indigenous yeasts to preserve the concept of `terroir`. The paper presents the study performed for the exploitation the microbial diversity in the Pietroasa wine region in Romania and to obtain autochthones yeasts biomass. Yeasts were isolated from grapes and must from Feteasca regala FR) and Cabernet Sauvignon (CS) varieties cultivated at the Pietroasa Viticulture and Vinification Research and Development Station. Five yeast isolates were selected and identified by the MALDI-TOF, these belonging to the S. cerevisiae and non-Saccharomyces genera. By using these yeast isolates, active dry biomass was obtained through fermentation on a substrate based on sterilised diluted must (from the Feteasca regala and Cabernet Sauvignon varieties), synthetic sterilised media, followed by freeze-drying. Subsequently, the active dry biomass was used to obtain wines, using two types of nutrients: ET (Energyvin Thiols) and NO (Nutristart® ORG). All studied yeast isolates showed a promising potential for obtaining white wine from Feteasca regala grapes and rosé wine from the Cabernet Sauvignon grapes variety.
38
Guo, Y. Z., C. Piras, A. Soggiu, M. Chanrot, R. Båge, G. Andersson, P. Reinaud, et al. "159 CHANGES IN PROTEIN EXPRESSION PROFILES IN BOVINE ENDOMETRIAL EPITHELIAL CELLS (bEEC) FOLLOWING E. COLI LIPOPOLYSACCHARIDE CHALLENGE." Reproduction, Fertility and Development 27, no. 1 (2015): 170. http://dx.doi.org/10.1071/rdv27n1ab159.
Анотація:
E. coli is one of the most frequent bacteria involved in uterine diseases. Lipopolysaccharide (LPS) is a component of the outer membrane of Gram-negative bacteria involved in the pathogenic processes leading to postpartum metritis and endometritis in cattle. It also causes inflammation of the endometrium. Increase of cell proliferation by LPS is part of the inflammatory process and has been reported in human epithelial and immune cells (Martin et al. 2000 J. Immunol. 165, 139–147) and from bovine endometrial epithelial cells (bEEC) (Guo et al. 2014 Reprod. Fertil. Dev. 26, 165–166). The aim of this study was to investigate possible changes in protein expression in relation with the proliferative response of bEEC after challenge with E. coli-LPS. In vitro culture of bEEC was performed from 3 cows. On passage 5, bEEC from each individual were exposed to 0, 8, and 16 µg mL–1 LPS for 72 h. At time 0 and 72 h later, attached cells were counted and for each time and LPS dosage, cells were frozen for proteomic analyses. The variation of cells number over time was analysed by ANOVA (SAS 9.1, proc GLM; SAS Institute, Inc., Cary, NC, USA). All samples were analysed (every sample run in triplicate) by 2-D gel electrophoresis coupled to matrix-assisted laser desorption/ionization-time-of-flight (MALDI-TOF)/time-of-flight (TOF) mass spectrometry (MS) and shotgun nLC-MS/MS analysis. As reported before, a significant increase in cell number was observed for cells treated with 8 µg mL–1 LPS (P ≤ 0.001), whereas changes in cell number were highly variable and nonsignificant for 16 µg mL–1 LPS. From each sample, ~800 proteins were visualised. Results from 2-D gel coupled to MALDI-TOF/TOF were very reproducible (same responses between individual cows) and revealed changes in protein profiles very much related (from P < 0.05 to P < 0.01) to proliferative phenotypes for seven proteins. From shotgun analysis, 27 proteins were found significantly differentially expressed (P < 0.05 to P < 0.01) following exposure to LPS (21 up-regulated and 6 down-regulated). Among the 21 found as up-regulated, 20 were differentially expressed both for the 8 and 16 µg mL–1 LPS, whereas 5 out of 6 were down-regulated for both dosages. Differentially expressed proteins were associated to cell proliferation, apoptosis, oxidative stress, regulation of histones, allergy, and general cell metabolism pathways. Candidate proteins need to be confirmed from larger series of individuals and relevant pathways further studied. Research was partially funded by RMUSTV.
39
Bukharin, O. V., O. E. Chelpachenko, E. I. Danilova, I. N. Chainikova, N. B. Perunova, E. V. Ivanova, I. A. Nikiforov, L. P. Fedotova, T. A. Bondarenko, and A. V. Salgina. "GUT MICROSYMBIOCENOSIS IN CHILDREN WITH REACTIVE ARTHRITIS." Journal of microbiology, epidemiology and immunobiology, no. 6 (December 28, 2016): 41–48. http://dx.doi.org/10.36233/0372-9311-2016-6-41-48.
Анотація:
Aim. To study the state of gut microsymbiocenosis in children with reactive arthritis (RA), with the assessment of biofilm formation (BFF) of microsymbionts and the ability to change cytokine levels (their anticytokine activity) in vitro. Materials and methods. The investigation of gut microsymbiocenosis by means of bacteriological method was conducted in 34 children with RA and 25 relatively healthy 3 - 16 year- old children. Microorganisms were identified with the help of MALDI-TOF mass-spectrometry, anticytokine activity (АСА) of microsymbionts - according to Bukharin O.V. et al. (2011), biofilm formation - according to O’Toole G.A., Kolter R. (1998). Results. On the ground of species composition differences of gut microbiota discrimination model was created which allowed to separate the group of children with RA from healthy individuals. Microsymbiocenosis of patients with RAwas characterized by increasing number of opportunistic microorganisms (OM) (enterobacteria, clostridia, bacteroides, and Candida), BFF and АСА level. Conclusion. The obtained data greatly contribute to the deciphering of spondylo-arthritis and disclose the role of microbial factor under given pathology. Hypercolonisation of human gut with OM, having pronounced ability to BFF and regulating cytokine level, promotes strengthening of arthritogenic potential and serves as additional marker of arthritis development risk in children.
40
Metaane, S., P. R. Burgel, D. Hubert, S. Bruel, B. Dauphin, and A. Paugam. "Identification par spectrométrie de masse de type MALDI-TOF de différentes espèces de Scedosporium isolées d’expectorations de patients adultes mucoviscidosiques et non mucoviscidosiques." Journal de Mycologie Médicale 24, no. 1 (March 2014): 75. http://dx.doi.org/10.1016/j.mycmed.2014.01.050.
41
Kharseeva, Galina G., E. O. Mangutov, O. M. But, A. V. Chepusova, and E. L. Alutina. "ANALYSIS OF THE FREQUENCY OF ISOLATION OF CORYNEBACTERIA NON-DIPHTHERIA FROM PATIENTS WITH INFLAMMATORY DISEASES OF THE RESPIRATORY TRACT." Russian Clinical Laboratory Diagnostics 64, no. 7 (October 7, 2019): 430–34. http://dx.doi.org/10.18821/0869-2084-2019-64-7-430-434.
Анотація:
Corynebacteria non-diphtheria and, in particular, C. pseudodiphtheriticum species that are closely related to C. propinquum and C. striatum form a group of new respiratory pathogens leading to the development of bronchitis, tracheitis, exacerbation of chronic obstructive pulmonary diseases, nosocomial pneumonia and other pathology. The goal is to analyze the frequency of the release of Сorynebacteria non-diphtheria from the upper respiratory tract of patients with various inflammatory diseases of the respiratory tract. Strains of Сorynebacteria non-diphtheria (C. pseudodiphtheriticum, C. propinquum, C. accolens, et al.), isolated from patients with inflammatory diseases of the respiratory tract (60 pcs.) and practically healthy individuals (31 pcs.) were studied. Identification of Сorynebacteria was performed using the method of mass spectrometry (MALDI-ToFMS). Сorynebacteria non-diphtheria in the amount of 105 and higher were more frequently detected with the development of chronic tonsillitis (60.0%) and nasopharyngitis (30%). The strains of C. pseudodiphtheriticum (40.0±6.4%) and the closely related species C. propinquum (21.7±5.3%) were mainly found; much less often - C. accolens (8.3±3.6%), C. afermentans (6.7±3.3%), et al. In 86.7% of cases, Corynebacteria non-diphtheria were isolated from children. In chronic tonsillitis, C. pseudodiphtheriticum and the closely related species of C. propinquum were isolated more often; in nasopharyngitis and bronchitis - С. pseudodiphtheriticum. Isolation of Corynebacteria non-diphtheria and, especially, C. pseudodiphtheriticum, C. propinquum, C. accolens species from patients with inflammatory diseases of the respiratory tract in the amount of 105 and above, if there are no other pathogenic microorganisms in the role of microbial associates, of clinical importance.
42
Arshad, N., P. Bémer, G. Le Gargasson, A. Guillouzouic, and S. Corvec. "Identification incorrecte par MALDI-TOF VitekMS de Cutibacterium namnetense à Cutibacterium acnes parmi des isolats cliniques : utilité du séquençage gyrB et nouveau pathogène osseux ?" Médecine et Maladies Infectieuses 48, no. 4 (June 2018): S93. http://dx.doi.org/10.1016/j.medmal.2018.04.233.
43
Dekio, Itaru, Mitsuo Sakamoto, Tomo Suzuki, Masahiro Yuki, Shigeru Kinoshita, Yoshiyuki Murakami, and Moriya Ohkuma. "Cutibacterium modestum sp. nov., isolated from meibum of human meibomian glands, and emended descriptions of Cutibacterium granulosum and Cutibacterium namnetense." International Journal of Systematic and Evolutionary Microbiology 70, no. 4 (April 1, 2020): 2457–62. http://dx.doi.org/10.1099/ijsem.0.004058.
Анотація:
An anaerobic and aerotolerant bacterium, strain M12T, was isolated from the meibum of inflamed human meibomian glands. Cells of the strain was Gram-stain-positive, non-spore-forming and non-motile rods. Growth on trypticase soy agar plates supplemented with 5 % sheep blood was fastest at 30–37 °C under anaerobic conditions. The 16S rRNA gene sequence of the strain revealed that it belongs to the genus Cutibacterium with a 98.0 % similarity value to the closest species, Cutibacterium acnes . Genome analysis of the strain with type strains of the other Cutibacterium species resulted in digital DNA–DNA hybridization values of 32.3–22.3% and average nucleotide identity (OrthoANI) values of 86.7–73.6 %. Biochemical and physiological analyses using API rapid ID 32A and API Coryne kits revealed relatively low reactivity of the strain compared with C. acnes and Cutibacterium namnetense . The most abundant major cellular fatty acid was iso-C15 : 0. Fermentation end-products from glucose were propionate, lactate, succinate and acetate. The diagnostic diamino acid of the peptidoglycan was meso-diaminopimelic acid. Major menaquinones were MK-9(H4), MK-9(H2) and MK-9. The major peaks of the MALDI-TOF mass spectrometry spectrum were at 3493, 3712, 6986 and 7424 Da. The DNA G+C content was 59.9 mol%. Based on these findings, we propose a novel species, Cutibacterium modestum. The type strain of C. modestum is M12T (=JCM 33380T=DSM 109769T). On the basis of further genomic analysis, we also provide emended descriptions of Cutibacterium granulosum (Prévot 1938) Scholz and Kilian 2016 and Cutibacterium namnetense (Aubin et al. 2016) Nouioui et al. 2018.
44
Bossio, Sabrina, Laura De Stefano, Mariavaleria Pellicanò, Angela Palummo, Francesca Storino, Nadia Caruso, Giovanni Iaquinta, et al. "Differentially Expressed Protein Patterns in Chronic Lymphocytic Leukemia (CLL) after Thymosin beta4 (Tb4) and Lenalidomide (Len) Treatment: Two-Dimensional Gel Electrophoresis (2DE) Analysis." Blood 126, no. 23 (December 3, 2015): 1729. http://dx.doi.org/10.1182/blood.v126.23.1729.1729.
Анотація:
Abstract Proteomic approaches are commonly secondary to genetic studies but are essential in the multi-disciplinary field of hematological research. As opposed to mRNA microarray data, proteomics provides a better understanding of which proteins are actually expressed, although the identification of specific proteins remains challenging (Unwin et al Blood Rev 2014; Boyd J Proteomics 2010). In neoplastic hematology such as CLL, protein studies have contributed to the elucidation of disease mechanisms, defined prognostic or therapeutic biomarkers (Boyd J Proteomics 2010). In this study we used proteomics and 2DE analysis to evaluate differential protein expression patterns after treatment with Len. Len can improve immune dysfunction in CLL by repairing F-actin polymerization and signaling at the immunological synapse (Ramsay et al 2008 J Clin Invest). Our previous data obtained from MALDI-TOF analysis identified Tβ4, a G-actin sequestering protein involved in the regeneration of injured tissues and cell migration, as a downregulated protein in CLL patients, also confirmed by an independent GEP analysis comparing B-cell from CLL cases (n=80) and normal controls (n=6), supported by Tβ4 mRNA down-regulation in CLL (3604±1244 vs 5715±1004, respectively; mean±SD; p=0.001). Here, we investigated whether purified B-CLL cells respond differently to the chemoattractant SDF1a and whether different protein expression patterns can be identified after exogenous Tβ4 and Len treatment using 2DE analysis. Highly purified B-CLL lymphocytes were isolated from untreated Binet stage A CLL patients prospectively enrolled from diagnosis (O-CLL1 protocol, clinicaltrial.gov identifier NCT00917540) and healthy controls. Tb4 was identified by MALDI TOF using 100 patient samples. Next, cells were pre-treated with Len (5uM) and then treated with Tb4 (100nM) for 30min. Cells were plated in transwells using 5.0 um pores with SDF1a as chemoattractant for migration assays. Protein was extracted from CLL cell pellets by RIPA buffer and quantified. Sample preparation and 2DE was performed as described by Scielzo et al (J Clin Inves, 2005). Protein samples (100 ug) were applied to 7-cm IPG strips, pH 3-11NL (Amersham Biosciences), respectively, by in-gel rehydration. Isoelectric focusing was performed with a Protean i12 IEF system (Biorad). Strips were equilibrated and loaded onto 9-16% gradient acrylamide SDS-PAGE gels for the second dimension separation. Silver nitrate staining (Sinha P et al Proteomic, 2001) was used to visualize proteins and images were digitally acquired (ChemiDoc MP, Biorad) and spots were analyzed using PDQuest basic 2D Gel Analysis Software (Biorad). CLL samples with the lowest Tβ4 expression (n=12) also had higher F-actin levels as evaluated by FC analysis than normal controls. B-CLL strongly responded to the migratory stimulus SDF-1a, which was further increased (by 20%) in presence of Len treatment, likely due to an alteration in actin remodeling and changes in the expression of unknown proteins. Purified CD19+CD5+ leukemic cells were lysed and proteins resolved on 2DE and visualized by silver nitrate staining. The protein profile analysis on silver-stained gels showed a number of protein spots ranging from 18 to 60kDa that were differentially expressed with respect to untreated cells. Our preliminary qualitative analysis suggests that there are groups of proteins with a lower expression in the presence of Tβ4 or Len, which are more strongly inhibited following exposure to their combination. Conversely, an opposite pattern of protein expression was observed whereby an additive effect on protein expression was observed by combined exposure to Tβ4 and Len. This approach allowed us to identify an altered protein expression pattern after treatment with Len and Tβ4, and may be useful to identify changes in expression profiles of CLL proteins, which may translate into functional differences in the malignant clone. Disclosures No relevant conflicts of interest to declare.
45
Miles, Stephen M., Jan Freas, and Philip J. Fay. "Mapping Factor VIIIa Inter-Subunit Interactions Using Chemical Modification and Zero-Length Cross-Linking." Blood 106, no. 11 (November 16, 2005): 1016. http://dx.doi.org/10.1182/blood.v106.11.1016.1016.
Анотація:
Abstract Activated factor VIII (FVIIIa) is a complex of three subunits, designated A1, A2, and A3C1C2, that serves as a cofactor for factor IXa in the proteolytic activation of factor X. The structure and surface interactions between factor VIIIa subunits have yet to be fully defined, but a homology-based model is available as a predictive tool (Stoylova-McPhie et al, Blood, 2002). To investigate FVIIIa inter-subunit interactions we used a chemical modification approach to determine surface-exposed and potentially interactive residues. The protein modification reagents acetic anhydride and diethylpyrocarbonate (DEPC), that modify lysine and histidine residues, respectively, were used to identify residues that are modified in the free subunits but protected in the bound complex. The modified samples were digested with various proteases and the resulting digests were analyzed by MALDI-TOF mass spectrometry to determine sites of peptide modification (Figure 1). The majority of data observed was in agreement with the A1/A3C1C2 interface in the predicted model. Protection was observed at Lys-89, Lys-142, and Lys-230 in the A1 subunit, all of which are near the A3C1C2 interface. While most His residues were completely modified by DEPC, partial protection was seen at His-1954, His-1957, and His-1961 in A3C1C2, which we have previously shown to be an A1-interactive region (Ansong and Fay, Biochemistry, 2005). In addition to this data, we attempted to define inter-subunit contacts by covalently cross-linking the FVIIIa complex. Using the zero-length cross-linker 1-ethyl-3-[3-dimethylaminopropyl] carbodiimide (EDC), which cross-links Glu/Asp residues to Lys, four distinct cross-linked bands were identified by SDS-PAGE (Figure 2). The composition of the cross-linked products was determined using proteolytic digestion and MALDI analysis. The four products correspond to each possible combination of the three subunits. Interestingly, there are no candidate cross-link sites between A1 and A2 in the model, yet this cross-link is the most predominant of the four. One alternative is that the A1–A2 cross-link may involve the A1 337–372 region that is not represented in the model. Taken together, the data are directed toward physically defining interactive regions between FVIIIa subunits and serve to test and supplement current structure models. FIGURE 1, FIGURE 2 FIGURE 1, FIGURE 2.
46
Lacroix, C., A. Gicquel, B. Sendid, J. Meyerd, I. Accoceberry, N. François, F. Morio, et al. "Identification des levures du genre Candida par spectrométrie de masse de type MALDI-TOF : comparaison des systèmes Andromas ® et Bruker/Biotyper v.2 ®." Journal de Mycologie Médicale 22, no. 1 (March 2012): 107. http://dx.doi.org/10.1016/j.mycmed.2011.12.028.
47
Ansong, Charles, Stephen Miles, and Philip J. Fay. "Factor VIII A2 Domain Residues 497–510 and 584–593 Comprise the R8B12 Epitope." Blood 106, no. 11 (November 16, 2005): 4028. http://dx.doi.org/10.1182/blood.v106.11.4028.4028.
Анотація:
Abstract The murine monoclonal antibody R8B12 recognizes the C-terminal region (residues 563–740) of the A2 subunit of factor VIIIa (Fay et al. J. Biol. Chem. 266:20139, 1991), as judged by western blotting. The location of the R8B12 epitope within the A2 subunit is not known. In the present study we used affinity-directed mass spectrometry (Zhao et al. Proc. Natl. Acad. Sci. USA.93:4020, 1996) to map the R8B12 epitope. Purified A2 subunit was subjected to proteolytic digestion with trypsin. The digest was then subjected to immunoprecipitation (IP) using R8B12 IgG, in which peptide(s) are selected that bind the antibody. Subsequently the masses of the affinity-selected peptides were determined directly from the immune complex by MALDI-TOF mass spectrometry. Proteolysis of A2 with trypsin generated a pre-IP peptide fingerprint that covered ~40% of the A2 domain sequence (Figure, left). Analysis of the post-IP peptide fingerprint showed two masses, 1309 and 1654, as representing affinity-selected peptide fragments (Figure, right). A theoretical database search identified the 1309 mass peak as A2 domain residues 584–593 and the 1654 mass peak as A2 domain residues 497–510. Direct sequencing of both mass peaks confirmed the results of the theoretical database search. These residues mapped to regions on the A2 domain of the factor VIII A domain homology model that are surface exposed and proximal to each other. Taken together, the above results suggest that A2 domain residues 497–510 and 584–593 represent a discontinous epitope for R8B12. Furthermore, based upon blotting specificity of R8B12 to the C-terminal portion of A2, we speculate that the latter sequence that forms this epitope makes a substantially greater contribution to the binding energy. Figure Figure
48
Lali, S. E., B. Marchetti, and D. Govaerts. "Intégration et accréditation ISO 15189 du Bruker Microflextm LT/Biotypertm Maldi-Tof System au sein du laboratoire de microbiologie de l’hôpital André-Vésale, CHU de Charleroi." Immuno-analyse & Biologie Spécialisée 27, no. 5 (October 2012): 305–6. http://dx.doi.org/10.1016/j.immbio.2012.07.019.
49
Zhang, Sujiang, Jianyong Li, Weida Li, Junhong Song, Limin Duan, Mize Lu, and Hongxia Qiu. "The Investigation of JAK2 V617F Mutation in Chinese Patients with Hematological Malignancies." Blood 108, no. 11 (November 16, 2006): 4918. http://dx.doi.org/10.1182/blood.v108.11.4918.4918.
Анотація:
Abstract Myeloproliferative Diseases (MPD) are a spectrum of pathogenetically related disorders of varying clinical manifestations, characterized by neoplastic expansion of relatively mature granulocyte, erythroid, megakaryocyte, or monocyte and eosinocyte lineage cells. Recently a novel point mutation affecting the Janus tyrosine kinase 2 (JAK2 V617F) was identified as pathogenetically mechanisms by multiple competing groups. To investigate its prevalence and clinical significance in Chinese patients with hematological malignancies, we introduced Allele-specific PCR (AS-PCR) combined with sequence analysis to screen JAK2 V617F mutation. A total of 98 Chinese MPD patients and 120 additional hematological malignancies including AML, ALL, MDS were analyzed for the JAK2 V617F mutation. 98 MPD patients were referred for PV (n=57), ET (n=18), IMF (n=12), HES (n=2) and CML (n=9). 2 ml peripheral blood samples of MPD and 5–10 ml bone marrow samples of AML, ALL, MDS at the time of initial diagnosis were obtained with informed consent and genomic DNA was isolated presently. In addition, peripheral blood samples from 20 healthy donors were also collected as control. All samples were first screened by AS-PCR. The positive samples were subsequently confirmed by sequence analysis. The results showed that JAK2 V617F mutation was detected in 43 of 57 PV patients (75.4%), 7 of 18 ET patients (38.9%) and 5 of 12 IMF patients (41.7%). None of the AML, ALL, MDS, CML was found JAK2 V617F. There is no statistical difference of JAK2 V617F positive ratio between PV, ET and IMF. Furthermore, the mutation was not detected in the HES patient and 20 healthy controls. There is no other mutations and polymorphisms throughout exon 12 of JAK2. To our knowledge, this is the first report of JAK2 V617F mutation in a number of Chinese patients with hematological malignancies especially BCR/ABL-negative MPD. The incidence of JAK2 V617F of our study is a little lower compared with other publications especially in PV patients. The main reason may be firstly attributed to ethnic difference. In addition, with more MPD patients introduced and more sensitive methods such as ARMS-PCR or Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS) applied, more JAK2 V617F mutation will be identified.
50
Kwak, Jae-Yong, Na-Ri Lee, Eun-Kee Song, Chang-Yeol Yim, and Yong-Geun Kwak. "The Comparative Analysis of Serum Proteomes for the Discovery of Biomarkers for Essential Thrombocythemia." Blood 106, no. 11 (November 16, 2005): 4004. http://dx.doi.org/10.1182/blood.v106.11.4004.4004.
Анотація:
Abstract Background: Essential thrombocythemia(ET) is a clonal disorder involving a hematopoietic progenitor cell and is manifested by the overproduction of platelets. There is as yet no pathognomonic diagnostic test. In our study, to identify the biomarkers for an initial diagnosis by a lesser invasive method, serum proteins reflecting alteration their proteomes were analysed. Methods: We compared two-dimensional electrophoresis patterns of human sera of twelve patients with ET with that of normal twelve subjects. The differentially expressed spots were identified by matrix-assisted laser desorption/ionization-time-of flight(MALDI-TOF) and electrospray ionization quadupole time of flight(ESI-Q-TOF)mass spectrometry. Results: Twenty two spots that expressed differentially in ET group were found. The expression levels of fibrinogen γ, fibrinogen γ-A chain precursor, ALB protein, α-2 macroglobulin, H factor-1, α-2-macroglobulin precursor, ceruloplasmin, α-1-B-glycoprotein, α-2-plasmin inhibitor, α-1-B-glycoprotein and an unknown protein were up-regulated in serum of ET patients, whereas the other proteins including complement fator B preproprotein, afamin precursor, inter-α inhibitor H4, immunoglobulin heavy chain constant region γ 1, immunoglobulin κ light chain and an unknown protein down-regulated. Conclusion: These results suggest that these proteins can be used as less invasive diagnostic and follow-up biomarkers of ET if further studies were done. The proteins significantly changed in sera of ET Spot no. Name Accession no. % coverage Mw(KDa)/pI(Measured) Mw(KDa)/pI(database) ET(%) 67 fibrinogen γ 223170 39 50.57/5.6 46.82/5.5 2.54±0.52 69 ” 223170 33 50.57/5.6 46.83/5.5 3.93±1.19 73 fibrinogen γ-A chain precursor 71827 23 48.93/5.6 50.11/5.7 3.89±0.98 74 ” 71827 22 48.93/5.8 50.11/5.7 3.10±0.91 77 ALB protein 27692693 21 50.57/5.9 48.65/6.0 3.77±0.58 89 α-2-macroglobulin 177872 14 263.89/6.1 71.35/5.5 1.95±0.32 100 H factor 1 4504875 20 248.95/5.7 143.79/6.3 5.8±1.78 112 α-2-macroglobulin precursor 4557225 9 151.38/5.7 164.69/6.0 2.18±0.38 125 complement factor B preproprotein 4502397 14 151.38/5.7 86.86/6.6 0.49±0.07 138 afamin precursor 4501987 12 65.01/5.1 85.53/6.3 0.31±0.07 139 ” 4501987 12 65.01/5.1 85.53/6.3 0.36±0.09 140 inter- α inhibitor H4 31542984 12 93.33/5.3 103.57/6.5 0.36±0.08 154 ceruloplasmin 4557485 8 132.89/5.3 123.04/5.4 3.34±0.80 161 α-1-B-glycoprotein 69990 21 84.50/5.0 52.49/5.6 4.23±0.98 162 α-2-plasmin inhibitor 11386143 14 78.58/5.0 54.92/5.8 1.95±0.38 167 α-1-B-glycoprotein 69990 28 80.87/5.3 52.49/5.6 1.73±0.04 210 Not identified - - 38.47/4.6 - 2.77±0.20 249 immunoglobulin heavy chain constant regionγ 1 12054072 15 39.021/8.1 36.65/8.8 0.40±0.09 250 ” 12054072 18 38.79/8.2 36.65/8.8 0.34±0.14 252 ” 12054072 19 38.50/8.3 36.65/8.8 0.37±0.09 262 unknown 15679996 36 24.73/8.2 23.32/8.6 0.30±0.13 265 immunoglobulinκ light chain 2765423 22 25.75/8.8 25.93/9.2 0.28±0.12 Figure Figure