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https://www.readbyqxmd.com/read/29157597/donor-specific-hla-antibodies-in-predicting-crossmatch-outcome-comparison-of-three-different-laboratory-techniques
#1
J P Peräsaari, T Jaatinen, J Merenmies
The virtual crossmatch, which is based on single antigen bead technology, is used in the prediction of crossmatch results. However, this assay differs in sensitivity and specificity from crossmatch methods. In our study, the results of physical crossmatches, performed with three different methods, were assessed against virtual crossmatch results. The aim was to determine the potential cut-off values for donor specific antibodies (DSA) that would predict the crossmatch results obtained by different methods. The results of different crossmatch techniques were correlated with the virtual crossmatch...
November 17, 2017: Transplant Immunology
https://www.readbyqxmd.com/read/29157295/single-cell-multiplexed-cytokine-profiling-of-cd19-car-t-cells-reveals-a-diverse-landscape-of-polyfunctional-antigen-specific-response
#2
Qiong Xue, Emily Bettini, Patrick Paczkowski, Colin Ng, Alaina Kaiser, Timothy McConnell, Olja Kodrasi, Máire F Quigley, James Heath, Rong Fan, Sean Mackay, Mark E Dudley, Sadik H Kassim, Jing Zhou
BACKGROUND: It remains challenging to characterize the functional attributes of chimeric antigen receptor (CAR)-engineered T cell product targeting CD19 related to potency and immunotoxicity ex vivo, despite promising in vivo efficacy in patients with B cell malignancies. METHODS: We employed a single-cell, 16-plex cytokine microfluidics device and new analysis techniques to evaluate the functional profile of CD19 CAR-T cells upon antigen-specific stimulation. CAR-T cells were manufactured from human PBMCs transfected with the lentivirus encoding the CD19-BB-z transgene and expanded with anti-CD3/anti-CD28 coated beads...
November 21, 2017: Journal for Immunotherapy of Cancer
https://www.readbyqxmd.com/read/29149956/comparison-between-total-igg-c1q-and-c3d-single-antigen-bead-assays-in-detecting-class-i-complement-binding-anti-hla-antibodies
#3
M A Moreno Gonzales, D G Mitema, B H Smith, C A Schinstock, M D Stegall, L L Wakefield, N A Henderson, S R DeGoey, J D Kreuter, M J Gandhi
BACKGROUND: Complement-binding donor-specific antibodies (DSAs) are associated with antibody-mediated rejection and allograft loss. Novel single antigen bead (SAB) assays-that is, complement component 1q (C1q) and complement component 3d (C3d) assays-have been developed to specifically detect complement-binding DSA, but it remains unclear whether these assays have an improved ability to detect complement-binding DSA as compared with using the total IgG SAB assay with a high mean fluorescence intensity (MFI) cutoff...
November 2017: Transplantation Proceedings
https://www.readbyqxmd.com/read/29139297/direct-exosome-quantification-via-bivalent-cholesterol-labeled-dna-anchor-for-signal-amplification
#4
Fang He, Hui Liu, Xinggang Guo, Bin-Cheng Yin, Bang-Ce Ye
Exosomes, as an important subpopulation of extracellular vesicles (EVs), play an important role in intercellular communications in various important pathophysiological processes, especially cancer-related. However, reliable and convenient quantitative methods for their determination are still technically challenging. In this study, we developed an efficient and direct method by combining immunoaffinity and lipid membrane surface modification into a single platform for specific isolation and accurate quantification of exosomes...
November 15, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/29135920/environmental-burkholderia-cenocepacia-strain-enhances-fitness-by-serial-passages-during-long-term-chronic-airways-infection-in-mice
#5
Alessandra Bragonzi, Moira Paroni, Luisa Pirone, Ivan Coladarci, Fiorentina Ascenzioni, Annamaria Bevivino
Burkholderia cenocepacia is an important opportunistic pathogen in cystic fibrosis (CF) patients, and has also been isolated from natural environments. In previous work, we explored the virulence and pathogenic potential of environmental B. cenocepacia strains and demonstrated that they do not differ from clinical strains in some pathogenic traits. Here, we investigated the ability of the environmental B. cenocepacia Mex1 strain, isolated from the maize rhizosphere, to persist and increase its virulence after serial passages in a mouse model of chronic infection...
November 14, 2017: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/29125493/validation-of-ultrasensitive-mutant-huntingtin-detection-in-human-cerebrospinal-fluid-by-single-molecule-counting-immunoassay
#6
Valentina Fodale, Roberto Boggio, Manuel Daldin, Cristina Cariulo, Maria Carolina Spiezia, Lauren Mary Byrne, Blair R Leavitt, Edward Wild, Douglas Macdonald, Andreas Weiss, Alberto Bresciani
BACKGROUND: The measurement of disease-relevant biomarkers has become a major component of clinical trial design, but in the absence of rigorous clinical and analytical validation of detection methodology, interpretation of results may be misleading. In Huntington's disease (HD), measurement of the concentration of mutant huntingtin protein (mHTT) in cerebrospinal fluid (CSF) of patients may serve as both a disease progression biomarker and a pharmacodynamic readout for HTT-lowering therapeutic approaches...
November 9, 2017: Journal of Huntington's Disease
https://www.readbyqxmd.com/read/29124927/integrated-continuous-emulsion-creamer
#7
Wesley Cochrane, Amber L Hackler, Valerie J Cavett, Alexander K Price, Brian M Paegel
Reaction incubation is ubiquitous in high-throughput screening workflows, including those at the microfluidic droplet scale. Fully integrated microfluidic processors that generate, incubate, and sort droplets for continuous droplet screening must passively handle a flowing emulsion such that droplet-droplet incubation time variation is minimized. Here, we disclose an integrated microfluidic emulsion creamer that close packs assay droplets by draining away excess oil through microfabricated drain channels. The drained oil co-flows with creamed emulsion and then reintroduces the oil to disperse the droplets at the circuit terminus for analysis...
November 10, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/29122726/improvement-of-ubiquitylation-site-detection-by-orbitrap-mass-spectrometry
#8
Lennart van der Wal, Karel Bezstarosti, Karen A Sap, Dick H W Dekkers, Erikjan Rijkers, Edwin Mientjes, Ype Elgersma, Jeroen A A Demmers
Ubiquitylation is an important posttranslational protein modification that is involved in many cellular events. Immunopurification of peptides containing a K-ε-diglycine (diGly) remnant as a mark of ubiquitylation combined with mass spectrometric detection has resulted in an explosion of the number of identified ubiquitylation sites. Here, we present several significant improvements to this workflow, including fast, offline and crude high pH reverse-phase fractionation of tryptic peptides into only three fractions with simultaneous desalting prior to immunopurification and better control of the peptide fragmentation settings in the Orbitrap HCD cell...
November 6, 2017: Journal of Proteomics
https://www.readbyqxmd.com/read/29120349/magnetic-beads-based-sensor-with-tailored-sensitivity-for-rapid-and-single-step-amperometric-determination-of-mirnas
#9
Eva Vargas, Rebeca M Torrente-Rodríguez, Víctor Ruiz-Valdepeñas Montiel, Eloy Povedano, María Pedrero, Juan J Montoya, Susana Campuzano, José M Pingarrón
This work describes a sensitive amperometric magneto-biosensor for single-step and rapid determination of microRNAs (miRNAs). The developed strategy involves the use of direct hybridization of the target miRNA (miRNA-21) with a specific biotinylated DNA probe immobilized on streptavidin-modified magnetic beads (MBs), and labeling of the resulting heteroduplexes with a specific DNA-RNA antibody and the bacterial protein A (ProtA) conjugated with an horseradish peroxidase (HRP) homopolymer (Poly-HRP40) as an enzymatic label for signal amplification...
November 9, 2017: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/29109009/rapid-optimized-flow-cytometric-crossmatch-fcxm-assays-the-halifax-and-halifaster-protocols
#10
Robert S Liwski, Anna L Greenshields, David M Conrad, Cathi Murphey, Robert A Bray, Jorge Neumann, Howard M Gebel
The flow cytometric crossmatch (FCXM) assay, which detects the presence of donor specific HLA antibodies in patient sera, is a cornerstone of HLA compatibility testing. Since relatively long FCXM assay turnaround times may contribute to transplant delays and increased graft ischemia time, we developed and validated two modified crossmatch procedures, namely the Halifax and Halifaster FCXM protocols. These protocols reduce FCXM assay time >60% and simplify their set-up without compromising quality or sensitivity...
November 3, 2017: Human Immunology
https://www.readbyqxmd.com/read/29108474/molecular-testing-for-oncogenic-gene-alterations-in-pediatric-thyroid-lesions
#11
Sogol Mostoufi-Moab, Emmanuel Labourier, Lisa Sullivan, Virginia A LiVolsi, Yimei Li, Rui Xiao, Sylvie Beaudenon-Huibregtse, Ken Kazahaya, N Scott Adzick, Zubair W Baloch, Andrew J Bauer
BACKGROUND: Thyroid nodules are less common in pediatric patients (≤18 years) than in adults. The Bethesda System for Reporting Thyroid Cytopathology allows for individual risk stratification, but a significant number of nodules are indeterminate. Incorporating gene mutation panels and gene expression classifiers may aid in pre-operative diagnosis. The overall aim of this study was to assess the prevalence of oncogene alterations in a representative pediatric population and across a broad-spectrum of thyroid tumor diagnoses...
November 6, 2017: Thyroid: Official Journal of the American Thyroid Association
https://www.readbyqxmd.com/read/29104846/single-molecule-interactions-of-a-monoclonal-anti-dna-antibody-with-dna
#12
Tatiana A Nevzorova, Qingze Zhao, Yakov A Lomakin, Anastasia A Ponomareva, Alexander R Mukhitov, Prashant K Purohit, John W Weisel, Rustem I Litvinov
Interactions of DNA with proteins are essential for key biological processes and have both a fundamental and practical significance. In particular, DNA binding to anti-DNA antibodies is a pathogenic mechanism in autoimmune pathology, such as systemic lupus erythematosus. Here we measured at the single-molecule level binding and forced unbinding of surface-attached DNA and a monoclonal anti-DNA antibody MRL4 from a lupus erythematosus mouse. In optical trap-based force spectroscopy, a microscopic antibodycoated latex bead is trapped by a focused laser beam and repeatedly brought into contact with a DNA-coated surface...
March 2017: BioNanoScience
https://www.readbyqxmd.com/read/29101041/intramolecular-structural-parameters-are-key-modulators-of-the-gel-liquid-transition-in-coarse-grained-simulations-of-dppc-and-dopc-lipid-bilayers
#13
S Jaschonek, M Cascella, J Gauss, G Diezemann, G Milano
The capability of coarse-grained models based on the MARTINI mapping to reproduce the gel-liquid phase transition in saturated and unsaturated model lipids was investigated. We found that the model is able to reproduce a lower critical temperature for 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) with respect to 1,2-dipalmitoyl-sn-glycero-3-phosphocholin (DPPC). Nonetheless, the appearance of a gel phase for DOPC is strictly dependent on the intramolecular parameters chosen to model its molecular structure...
October 31, 2017: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/29083305/single-molecule-force-spectroscopy-of-protein-membrane-interactions
#14
Lu Ma, Yiying Cai, Yanghui Li, Junyi Jiao, Zhenyong Wu, Ben O'Shaughnessy, Pietro De Camilli, Erdem Karatekin, Yongli Zhang
Many biological processes rely on protein-membrane interactions in the presence of mechanical forces, yet high resolution methods to quantify such interactions are lacking. Here, we describe a single-molecule force spectroscopy approach to quantify membrane binding of C2 domains in Synaptotagmin-1 (Syt1) and Extended Synaptotagmin-2 (E-Syt2). Syts and E-Syts bind the plasma membrane via multiple C2 domains, bridging the plasma membrane with synaptic vesicles or endoplasmic reticulum to regulate membrane fusion or lipid exchange, respectively...
October 30, 2017: ELife
https://www.readbyqxmd.com/read/29082223/healthcare-costs-of-transarterial-chemoembolization-in-the-treatment-of-hepatocellular-carcinoma
#15
Waleed Fateen, Farooq Khan, Richard J O'NeiSSll, Martin W James, Stephen D Ryder, Guruprasad P Aithal
BACKGROUND: A meta-analysis comparing drug-eluting beads transarterial chemoembolization (DEB-TACE) with conventional transarterial chemoembolization (cTACE) has recently been published. On balance, no significant differences were found in terms of objective response and overall survival. The impact on healthcare costs had been studied in small series based on a hypothetical model and was in favor of DEB-TACE. We aimed to evaluate and compare health-care costs and effectiveness of both modalities in a cohort of patients from Nottingham, UK...
2017: Journal of Hepatocellular Carcinoma
https://www.readbyqxmd.com/read/29079485/single-shot-dual-wavelength-interferometric-microscopy
#16
Poorya Hosseini, Di Jin, Zahid Yaqoob, Peter T C So
Interferometric microscopy (IM) can provide complex field information of the biological samples with high spatial and temporal resolution with virtually no photodamage. Measuring wavelength-dependent information in particular has a wide range of applications from cell and tissue refractometry to the cellular biophysical measurements. IM measurements at multiple wavelengths are typically associated with a loss in temporal resolution, field of view, stability, sensitivity, and may involve using expensive equipment such as tunable filters or spatial light modulators...
October 24, 2017: Methods: a Companion to Methods in Enzymology
https://www.readbyqxmd.com/read/29075627/characteristics-of-donor-specific-antibodies-associated-with-antibody-mediated-rejection-in-lung-transplantation
#17
Antoine Roux, Ines Bendib Le Lan, Sonia Holifanjaniaina, Kimberly A Thomas, Clément Picard, Dominique Grenet, Sandra De Miranda, Benoit Douvry, Laurence Beaumont-Azuar, Edouard Sage, Jérôme Devaquet, Elise Cuquemelle, Morgan Le Guen, Caroline Suberbielle, Chantal Gautreau, Marc Stern, Maura Rossetti, Abdul Monem Hamid, Francois Parquin
Although donor-specific anti-human leukocyte antigen (HLA) antibodies (DSAs) are frequently found in recipients after lung transplantation (LT), the characteristics of DSA which influence antibody-mediated rejection (AMR) in LT are not fully defined. We retrospectively analyzed 206 consecutive LT patients of our center (2010-2013). DSAs were detected by using luminex single antigen beads assay and mean fluorescence intensity was assessed. Within the study population, 105 patients had positive DSA. Patients with and without AMR (AMR(Pos), n = 22, and AMR(Neg), n = 83, respectively) were compared...
2017: Frontiers in Medicine
https://www.readbyqxmd.com/read/29071674/mass-cytometry-assays-for-antigen-specific-t-cells-using-cytof
#18
Dongxia Lin, Holden T Maecker
T Cells specific for a single antigen tend to be rare, even after expansion of memory cells. They are commonly detected by in vitro stimulation with peptides or protein, followed by staining for intracellular cytokines. In this protocol, we use CyTOF(®) mass cytometry to collect single-cell data on a large number of cytokines/chemokines, as well as cell-surface proteins that characterize T cells and other immune cells. We also include a method for magnetic bead enrichment of antigen-stimulated T cells, based on their expression of CD154 and CD69...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/29063856/does-neoadjuvant-doxorubicin-drug-eluting-bead-transarterial-chemoembolization-improve-survival-in-patients-undergoing-liver-transplant-for-hepatocellular-carcinoma
#19
Dimitri Dorcaratto, Venkatesha Udupa, Niamh M Hogan, David P Brophy, Jeffrey W McCann, Donal Maguire, Justin Geoghegan, Colin P Cantwell, Emir Hoti
PURPOSE: We aimed to compare the overall (OS) and disease-free survival (DFS) of patients undergoing orthotopic liver transplant (OLT) for hepatocellular carcinoma who did and did not have neoadjuvant doxorubicin drug-eluting bead transarterial chemoembolization (DEB-TACE). METHODS: This is a retrospective study of 94 patients with HCC transplanted between 2000 and 2014 in a single tertiary center. Pre- and postoperative features, DFS and OS were compared between patients who received pre-OLT DEB-TACE (n=34, DEB-TACE group) and those who did not (n=60, non-TACE group)...
November 2017: Diagnostic and Interventional Radiology: Official Journal of the Turkish Society of Radiology
https://www.readbyqxmd.com/read/29059522/impact-of-microplastic-beads-and-fibers-on-waterflea-ceriodaphnia-dubia-survival-growth-and-reproduction-implications-of-single-and-mixture-exposures
#20
Shima Ziajahromi, Anupama Kumar, Peta A Neale, Frederic D L Leusch
There is limited knowledge regarding the adverse effects of wastewater-derived microplastics, particularly fibers, on aquatic biota. In this study, we examined the acute (48 h) and chronic (8 d) effects of microplastic polyester fibers and polyethylene (PE) beads on freshwater zooplankton Ceriodaphnia dubia. We also assessed the acute response of C. dubia to a binary mixture of microplastic beads and fibers for the first time. Acute exposure to fibers and PE beads both showed a dose-dependent effect on survival...
November 3, 2017: Environmental Science & Technology
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