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https://www.readbyqxmd.com/read/27909706/out-of-plane-integration-of-a-multimode-optical-fiber-for-single-particle-cell-detection-at-multiple-points-on-a-microfluidic-device-with-applications-to-particle-cell-counting-velocimetry-size-discrimination-and-the-analysis-of-single-cell-lysate-injections
#1
Jalal Sadeghi, Damith E W Patabadige, Anne H Culbertson, Hamid Latifi, Christopher T Culbertson
In this paper a single particle/cell-tracking microfluidic device that integrates an out-of-plane multimode optical fiber (OP-MMF) is reported. This OP-MMF is used to generate three excitation light-lines and three detection spots using only one excitation source and one detector. It takes advantage of an optical tunneling mode to create two excitation lines in a microfluidic channel emanating from a single fiber end. This method was used to accurately count particles/cells and perform velocity measurements and size discrimination...
December 2, 2016: Lab on a Chip
https://www.readbyqxmd.com/read/27904609/deciphering-biological-characteristics-of-tumorigenic-subpopulations-in-human-colorectal-cancer-reveals-cellular-plasticity
#2
Hamed Mirzaei, Hossein Salehi, Amirhossein Sahebkar, Amir Avan, Mahmoud Reza Jaafari, Afshin Namdar, Abbas Rezaei, Hamid Reza Mirzaei
BACKGROUND: It is supposed that human colorectal cancer consists of a phenotypically distinct population of tumorigenic cancer cells known as cancer stem cells (CSCs) which play a pivotal role in cancer progression, maintenance, metastasis, and the relapse. The aim of this effort was to investigate and compare biological characterizations of CD133(+) with CD133(-) cell subsets isolated from both primary and metastatic human colorectal tumors. MATERIALS AND METHODS: Using our optimized protocols, unfixed colorectal tumors were enzymatically and mechanically dissociated into single cells followed by evaluation of postdigestion viability...
2016: Journal of Research in Medical Sciences: the Official Journal of Isfahan University of Medical Sciences
https://www.readbyqxmd.com/read/27896766/multiplex-immunoassay-profiling
#3
Laurie Stephen
Multiplex immunoassays allow for the rapid profiling of biomarker proteins in biological fluids, using less sample and labor than single immunoassays. This chapter details the methods to develop and manufacture multiplex assays for the Luminex(®) platform. Although assay development is not included here, the same methods can be used to covalently couple antibodies to the Luminex beads and to label antibodies for the screening of sandwich pairs, if needed. The assay optimization, detection of cross-reactivity, and minimizing antibody interactions and matrix interferences will be addressed...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27894836/use-of-complement-binding-assays-to-assess-the-efficacy-of-antibody-mediated-rejection-therapy-and-prediction-of-graft-survival-in-kidney-transplantation
#4
Daniel S Ramon, Yihung Huang, Lili Zhao, TrisAnn Rendulic, Jeong M Park, Randall S Sung, Milagros Samaniego
BACKGROUND: The Luminex(®) single antigen bead assay (SAB) is the method of choice for monitoring the treatment for antibody-mediated rejection (AMR). A ⩾50% reduction of the dominant donor-specific antibody (IgG-DSA) mean fluorescence intensity (MFI) has been associated with improved kidney allograft survival, and C1q-fixing DSA activity is associated with poor outcomes in patients with AMR. We aimed to investigate if C1q-DSA can be used as a reliable predictor of response to therapy and allograft survival in patients with biopsy-proven AMR...
November 25, 2016: Human Immunology
https://www.readbyqxmd.com/read/27889122/genome-wide-association-study-for-cheese-yield-and-curd-nutrient-recovery-in-dairy-cows
#5
C Dadousis, S Biffani, C Cipolat-Gotet, E L Nicolazzi, G J M Rosa, D Gianola, A Rossoni, E Santus, G Bittante, A Cecchinato
Cheese production and consumption are increasing in many countries worldwide. As a result, interest has increased in strategies for genetic selection of individuals for technological traits of milk related to cheese yield (CY) in dairy cattle breeding. However, little is known about the genetic background of a cow's ability to produce cheese. Recently, a relatively large panel (1,264 cows) of different measures of individual cow CY and milk nutrient and energy recoveries in the cheese (REC) became available...
November 23, 2016: Journal of Dairy Science
https://www.readbyqxmd.com/read/27887864/optimization-of-cgmp-purification-and-expansion-of-umbilical-cord-blood-derived-t-regulatory-cells-in-support-of-first-in-human-clinical-trials
#6
David H McKenna, Darin Sumstad, Diane M Kadidlo, Bjorn Batdorf, Colin J Lord, Sarah C Merkel, Christine M Koellner, Julie M Curtsinger, Carl H June, James L Riley, Bruce L Levine, Jeffrey S Miller, Claudio G Brunstein, John E Wagner, Bruce R Blazar, Keli L Hippen
BACKGROUND AIMS: Thymic-derived regulatory T cells (tTreg) are critical regulators of the immune system. Adoptive tTreg transfer is a curative therapy for murine models of autoimmunity, graft rejection, and graft-versus-host disease (GVHD). We previously completed a "first-in-human" clinical trial using in vitro expanded umbilical cord blood (UCB)-derived tTreg to prevent GVHD in patients undergoing UCB hematopoietic stem cell transplantation (HSCT). tTreg were safe and demonstrated clinical efficacy, but low yield prevented further dose escalation...
November 22, 2016: Cytotherapy
https://www.readbyqxmd.com/read/27869283/controlling-the-electrical-conductive-network-formation-of-polymer-nanocomposites-via-polymer-functionalization
#7
Yangyang Gao, Youping Wu, Jun Liu, Liqun Zhang
By adopting coarse-grained molecular dynamics simulations, the effect of polymer functionalization on the relationship between the microstructure and the electric percolation probability of nanorod filled polymer nanocomposites has been investigated. At a low chain functionalization degree, the nanorods in the polymer matrix form isolated aggregates with a local order structure. At a moderate chain functionalization degree, the local order structure of the nanorod aggregate is gradually broken up. Meanwhile, excessive functionalization chain beads can connect the isolated aggregates together, which leads to the maximum size of nanorod aggregation...
November 21, 2016: Soft Matter
https://www.readbyqxmd.com/read/27866824/detection-of-her2-overexpressing-cancer-cells-using-keyhole-shaped-chamber-array-employing-a-magnetic-droplet-handling-system
#8
Mina Okochi, Shinji Koike, Masayoshi Tanaka, Hiroyuki Honda
An on-chip gene expression analysis compartmentalized in droplets was developed for detection of cancer cells at a single-cell level. The chip consists of a keyhole-shaped reaction chamber with hydrophobic modification employing a magnetic bead-droplet-handling system with a gate for bead separation. Using three kinds of water-based droplets in oil, a droplet with sample cells, a lysis buffer with magnetic beads, and RT-PCR buffer, parallel magnetic manipulation and fusion of droplets were performed using a magnet-handling device containing small external magnet patterns in an array...
November 11, 2016: Biosensors & Bioelectronics
https://www.readbyqxmd.com/read/27863041/immobilized-hematopoietic-growth-factors-onto-magnetic-particles-offer-a-scalable-strategy-for-cell-therapy-manufacturing-in-suspension-cultures
#9
Matthew J Worrallo, Rebecca L L Moore, Katie E Glen, Robert J Thomas
Hematopoietic therapies require high cell dosages and precise phenotype control for clinical success; scalable manufacturing processes therefore need to be economic and controllable, in particular with respect to culture medium and growth factor (GF) strategy. The aim of this work was to demonstrate the biological function, and integration within scalable systems, of a highly controllable immobilized growth factor (iGF) approach. GFs were biotinylated and attached to streptavidin coated magnetic particles. GF concentration during biotinylation, GF-biotin ratio, and GF lysine content were shown to control iGF surface concentration and enable predictable co-presentation of multiple GF on a single bead...
November 11, 2016: Biotechnology Journal
https://www.readbyqxmd.com/read/27862486/characterisation-of-maternal-human-leukocyte-antigen-class-i-antibodies-in-suspected-foetal-and-neonatal-alloimmune-thrombocytopenia
#10
E Refsum, A Mörtberg, J Dahl, S Meinke, M-K Auvinen, M Westgren, M Reilly, P Höglund, A Wikman
OBJECTIVES: To investigate the specificities and level of HLA class I antibodies in selected cases referred for suspected foetal and neonatal alloimmune thrombocytopenia (FNAIT). BACKGROUND: FNAIT occurs in 1 : 1-2000 live births, whereas maternal immunisation against human leukocyte antigen (HLA) class I is common. Whether HLA class I antibodies alone can cause FNAIT is debatable. MATERIAL AND METHODS: A total of 260 patient samples were referred between 2007 and 2012...
November 9, 2016: Transfusion Medicine
https://www.readbyqxmd.com/read/27856136/korean-multicenter-registry-of-transcatheter-arterial-chemoembolization-with-drug-eluting-embolic-agents-for-nodular-hepatocellular-carcinomas-six-month-outcome-analysis
#11
Myungsu Lee, Jin Wook Chung, Kwang-Hun Lee, Jong Yun Won, Ho Jong Chun, Han Chu Lee, Jin Hyoung Kim, In Joon Lee, Saebeom Hur, Hyo-Cheol Kim, Yoon Jun Kim, Gyoung Min Kim, Seung-Moon Joo, Jung Suk Oh
PURPOSE: To assess the efficacy and safety of transcatheter arterial chemoembolization with drug-eluting embolic (DEE) agents for nodular hepatocellular carcinoma (HCC). MATERIALS AND METHODS: The study design was a prospective multicenter registry-based, single-arm clinical trial that included 152 patients. One hundred three (67.8%) had a Child-Pugh class/score of A5, 114 (75.0%) had a performance status of 0, and 77 (50.7%) had Barcelona Clinic Liver Cancer (BCLC) stage A disease...
November 14, 2016: Journal of Vascular and Interventional Radiology: JVIR
https://www.readbyqxmd.com/read/27853145/marco-variants-are-associated-with-phagocytosis-pulmonary-tuberculosis-susceptibility-and-beijing-lineage
#12
N T T Thuong, T T B Tram, T D Dinh, P V K Thai, D Heemskerk, N D Bang, T T H Chau, D G Russell, G E Thwaites, T R Hawn, M Caws, S J Dunstan
Macrophage receptor with collagenous structure (MARCO) has an important role in the phagocytosis of Mycobacterium tuberculosis (M. tuberculosis). We hypothesized that MARCO polymorphisms are associated with phagocytosis, tuberculosis (TB) disease susceptibility and presentation, and infecting lineage. We used a human cellular model to examine how MARCO genotype mediates the immune response; a case-control study to investigate tuberculosis host genetic susceptibility; and a host-pathogen genetic analysis to study host-pathogen interactions...
December 2016: Genes and Immunity
https://www.readbyqxmd.com/read/27844439/simultaneous-manipulation-and-super-resolution-fluorescence-imaging-of-individual-kinetochores-coupled-to-microtubule-tips
#13
Yi Deng, Charles L Asbury
Kinetochores are large multiprotein complexes that drive mitotic chromosome movements by mechanically coupling them to the growing and shortening tips of spindle microtubules. Kinetochores are also regulatory hubs, somehow sensing when they are erroneously attached and, in response, releasing their incorrect attachments and generating diffusible wait signals to delay anaphase until proper attachments can form. The remarkable ability of a kinetochore to sense and respond to its attachment status might stem from attachment- or tension-dependent changes in the structural arrangement of its core subcomplexes...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27844437/observing-single-rna-polymerase-molecules-down-to-base-pair-resolution
#14
Anirban Chakraborty, Cong A Meng, Steven M Block
During transcriptional elongation, RNA polymerases (RNAP) employ a stepping mechanism to translocate along the DNA template while synthesizing RNA. Optical trapping assays permit the progress of single molecules of RNA polymerase to be monitored in real time, at resolutions down to the level of individual base pairs. Additionally, optical trapping assays permit the application of exquisitely controlled, external forces on RNAP. Responses to such forces can reveal details of the load-dependent kinetics of transcriptional elongation and pausing...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27844436/single-molecule-protein-folding-experiments-using-high-precision-optical-tweezers
#15
Junyi Jiao, Aleksander A Rebane, Lu Ma, Yongli Zhang
How proteins fold from linear chains of amino acids to delicate three-dimensional structures remains a fundamental biological problem. Single-molecule manipulation based on high-resolution optical tweezers (OT) provides a powerful approach to study protein folding with unprecedented spatiotemporal resolution. In this method, a single protein or protein complex is tethered between two beads confined in optical traps and pulled. Protein unfolding induced by the mechanical force is counteracted by the spontaneous folding of the protein, reaching a dynamic equilibrium at a characteristic force and rate...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27844425/beyond-the-hookean-spring-model-direct-measurement-of-optical-forces-through-light-momentum-changes
#16
Arnau Farré, Ferran Marsà, Mario Montes-Usategui
The ability to measure forces in the range of 0.1-100 pN is a key feature of optical tweezers used for biophysical and cell biological studies. Analysis of the interactions between biomolecules and the forces that biomolecular motors generate at the single-molecule level has provided valuable insights in the molecular mechanisms that govern key cellular functions such as gene expression and the long-distance transport of organelles. Methods for determining the minute forces that biomolecular motors generate exhibit notable constraints that limit their application for studies other than the well-controlled in vitro experiments (although recent advances have been made that permit more quantitative optical tweezers studies insight living cells)...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27835944/optimization-and-evaluation-of-luminex-performance-with-supernatants-of-antigen-stimulated-peripheral-blood-mononuclear-cells
#17
Mathieu Surenaud, Céline Manier, Laura Richert, Rodolphe Thiébaut, Yves Levy, Sophie Hue, Christine Lacabaratz
BACKGROUND: The Luminex bead-based multiplex assay is useful for quantifying immune mediators such as cytokines and chemokines. Cross-comparisons of reagents for this technique from different suppliers have already been performed using serum or plasma but rarely with supernatants collected from antigen-stimulated peripheral blood mononuclear cells (PBMC). Here, we first describe an optimization protocol for cell culture including quantity of cells and culture duration to obtain reproducible cytokine and chemokine quantifications...
November 11, 2016: BMC Immunology
https://www.readbyqxmd.com/read/27833183/couple-beads-an-integrated-method-of-natural-family-planning
#18
George Mulcaire-Jones, Richard J Fehring, Megan Bradshaw, Karen Brower, Gonzaga Lubega, Paskazia Lubega
Various fertility indicators are used by natural family planning methods to identify the fertile and infertile phases of a woman's menstrual cycle: mucus observations, cycle-day probabilities, basal body temperature readings, and hormonal measures of LH and estrogen. Simplified NFP methods generally make use of a single fertility indicator such as cycle-day probabilities (Standard Days Method) or mucus observations (Billings Ovulation Method). The Couple Bead Method integrates the two simplest fertility indicators, cycle-day probabilities and mucus observations, expanding its applicability to all women, regardless of cycle regularity and length...
February 2016: Linacre Quarterly
https://www.readbyqxmd.com/read/27829567/digital-quantification-of-gene-methylation-in-stool-dna-by-emulsion-pcr-coupled-with-hydrogel-immobilized-bead-array
#19
Yunlong Liu, Haiping Wu, Qiang Zhou, Qinxin Song, Jianzhong Rui, Bingjie Zou, Guohua Zhou
Aberrations of gene methylation in stool DNA (sDNA) is an effective biomarker for non-invasive colorectal cancer diagnosis. However, it is challenging to accurately quantitate the gene methylation levels in sDNA due to the low abundance and degradation of sDNA. In this study, a digital quantification strategy was proposed by combining emulsion PCR (emPCR) with hydrogel immobilized bead-array. The assay includes following steps: bisulfite conversion of sDNA, pre-amplification by PCR with specific primers containing 5' universal sequences, emPCR of pre-amplicons with beaded primers to achieve single-molecular amplification and identification of hydrogel embedding beads coated with amplicons...
October 22, 2016: Biosensors & Bioelectronics
https://www.readbyqxmd.com/read/27826602/a-novel-method-for-anti-hla-antibody-detection-using-personalized-peptide-arrays
#20
Pan Liu, Tomokazu Souma, Andrew Zu-Sern Wei, Xueying Xie, Xunrong Luo, Jing Jin
BACKGROUND: HLA mismatches are the primary cause of alloantibody-mediated rejection (AMR) in organ transplantation. To delineate antigenic and immunogenic potentials among individual HLA mismatches, information regarding antibody specificity at the epitope level, instead of the allelic level, is needed. METHODS: This study explores a direct screening method for HLA linear epitopes in kidney transplant patients. We custom synthesized a large panel of 15-residue HLA peptides in an array format and measured alloantibody reactivity to these peptides from the sera of post and/or pretransplant patients...
November 2016: Transplantation Direct
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