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Collagen high throughput

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https://www.readbyqxmd.com/read/28216326/a-scalable-lysyl-hydroxylase-2-expression-system-and-luciferase-based-enzymatic-activity-assay
#1
Hou-Fu Guo, Eun Jeong Cho, Ashwini K Devkota, Yulong Chen, William Russell, George N Phillips, Mitsuo Yamauchi, Kevin N Dalby, Jonathan M Kurie
Hydroxylysine aldehyde-derived collagen cross-links (HLCCs) accumulate in fibrotic tissues and certain types of cancer and are thought to drive the progression of these diseases. HLCC formation is initiated by lysyl hydroxylase 2 (LH2), an Fe(II) and α-ketoglutarate (αKG)-dependent oxygenase that hydroxylates telopeptidyl lysine residues on collagen. Development of LH2 antagonists for the treatment of these diseases will require a reliable source of recombinant LH2 protein and a non-radioactive LH2 enzymatic activity assay that is amenable to high throughput screens of small molecule libraries...
February 16, 2017: Archives of Biochemistry and Biophysics
https://www.readbyqxmd.com/read/28207883/application-of-an-optimized-flow-cytometry-based-quantification-of-platelet-activation-pact-monitoring-platelet-activation-in-platelet-concentrates
#2
Cécile H Kicken, Mark Roest, Yvonne M C Henskens, Bas de Laat, Dana Huskens
BACKGROUND: Previous studies have shown that flow cytometry is a reliable test to quantify platelet function in stored platelet concentrates (PC). It is thought that flow cytometry is laborious and hence expensive. We have optimized the flow cytometry-based quantification of agonist induced platelet activation (PACT) to a labor, time and more cost-efficient test. Currently the quality of PCs is only monitored by visual inspection, because available assays are unreliable or too laborious for use in a clinical transfusion laboratory...
2017: PloS One
https://www.readbyqxmd.com/read/28198402/an-integrated-microfluidic-device-for-rapid-and-high-sensitivity-analysis-of-circulating-tumor-cells
#3
Jianing Jiang, Hui Zhao, Weiliang Shu, Jing Tian, Yuqing Huang, Yongxin Song, Ruoyu Wang, Encheng Li, Dennis Slamon, Dongmei Hou, Xiaohui Du, Lichuan Zhang, Yan Chen, Qi Wang
Recently there has been a more focus on the development of an efficient technique for detection of circulating tumor cells (CTCs), due to their significance in prognosis and therapy of metastatic cancer. However, it remains a challenge because of the low count of CTCs in the blood. Herein, a rapid and high-sensitivity approach for CTCs detection using an integrated microfluidic system, consisting of a deterministic lateral displacement (DLD) isolating structure, an automatic purifying device with CD45-labeled immunomagnetic beads and a capturing platform coated with rat-tail collagen was reported...
February 15, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28181140/real-architecture-for-3d-tissue-raft%C3%A2-culture-system-improves-viability-and-maintains-insulin-and-glucagon-production-of-mouse-pancreatic-islet-cells
#4
Gabor J Szebeni, Zsuzsanna Tancos, Liliana Z Feher, Robert Alfoldi, Julianna Kobolak, Andras Dinnyes, Laszlo G Puskas
There is an unmet medical need for the improvement of pancreatic islet maintenance in culture. Due to restricted donor availability it is essential to ameliorate islet viability and graft engraftment. The aim of this study was to compare the standard tissue culture techniques with the advanced Real Architecture For 3D Tissue (RAFT™) culture system in terms of viability and hormone production. Here, we first report that islets embedded in RAFT™ collagen type I advanced tissue culture system maintain their tissue integrity better than in monolayer and suspension cultures...
February 8, 2017: Cytotechnology
https://www.readbyqxmd.com/read/28166248/characterization-of-three-dimensional-cancer-cell-migration-in-mixed-collagen-matrigel-scaffolds-using-microfluidics-and-image-analysis
#5
María Anguiano, Carlos Castilla, Martin Maška, Cristina Ederra, Rafael Peláez, Xabier Morales, Gorka Muñoz-Arrieta, Maite Mujika, Michal Kozubek, Arrate Muñoz-Barrutia, Ana Rouzaut, Sergio Arana, José Manuel Garcia-Aznar, Carlos Ortiz-de-Solorzano
Microfluidic devices are becoming mainstream tools to recapitulate in vitro the behavior of cells and tissues. In this study, we use microfluidic devices filled with hydrogels of mixed collagen-Matrigel composition to study the migration of lung cancer cells under different cancer invasion microenvironments. We present the design of the microfluidic device, characterize the hydrogels morphologically and mechanically and use quantitative image analysis to measure the migration of H1299 lung adenocarcinoma cancer cells in different experimental conditions...
2017: PloS One
https://www.readbyqxmd.com/read/28143748/a-three-dimensional-neural-spheroid-model-for-capillary-like-network-formation
#6
Molly E Boutin, Liana L Kramer, Liane L Livi, Tyler Brown, Christopher Moore, Diane Hoffman-Kim
BACKGROUND: In vitro three-dimensional neural spheroid models have an in vivo-like cell density, and have the potential to reduce animal usage and increase experimental throughput. The aim of this study was to establish a spheroid model to study the formation of capillary-like networks in a three-dimensional environment that incorporates both neuronal and glial cell types, and does not require exogenous vasculogenic growth factors. NEW METHOD: We created self-assembled, scaffold-free cellular spheroids using primary-derived postnatal rodent cortex as a cell source...
January 29, 2017: Journal of Neuroscience Methods
https://www.readbyqxmd.com/read/28129584/bone-natural-autofluorescence-and-confocal-laser-scanning-microscopy-preliminary-results-of-a-novel-useful-tool-to-distinguish-between-forensic-and-ancient-human-skeletal-remains
#7
Luigi Capasso, Ruggero D'Anastasio, Simone Guarnieri, Joan Viciano, Maria Mariggiò
The fast, high-throughput distinction between palaeoanthropological/archaeological remains and recent forensic/clinical bone samples is of vital importance in the field of medico-legal science. In this paper, a novel dating method was developed using the autofluorescence of human bones and the confocal laser scanning microscope as the means to distinguish between archaeological and forensic anthropological skeletal findings. Human bones exhibit fluorescence, typically induced by natural antibiotics that are absorbed by collagen, and provide secondary, exogenous fluorophores...
January 22, 2017: Forensic Science International
https://www.readbyqxmd.com/read/27989921/a-model-system-for-developing-a-tissue-engineered-meniscal-enthesis
#8
Mary Clare McCorry, Melissa M Mansfield, Xiaozhou Sha, Daniel J Coppola, Jonathan W Lee, Lawrence J Bonassar
: The meniscus acts as a stabilizer, lubricator, and load distributer in the knee joint. The mechanical stability of the meniscus depends on its connection to the underlying bone by a fibrocartilage to bone transition zone called the meniscal enthesis. Tissue engineered menisci hold great promise as a treatment alternative however lack a means of integrated fixation to the underlying bone needed in order for a tissue engineered meniscal replacement to be successful. Tissue engineering the meniscal enthesis is a difficult task given the complex gradients of cell type, mineral, and extracellular matrix molecules...
October 29, 2016: Acta Biomaterialia
https://www.readbyqxmd.com/read/27870847/creating-high-resolution-multiscale-maps-of-human-tissue-using-multi-beam-sem
#9
André F Pereira, Daniel J Hageman, Tomasz Garbowski, Christof Riedesel, Ulf Knothe, Dirk Zeidler, Melissa L Knothe Tate
Multi-beam scanning electron microscopy (mSEM) enables high-throughput, nano-resolution imaging of macroscopic tissue samples, providing an unprecedented means for structure-function characterization of biological tissues and their cellular inhabitants, seamlessly across multiple length scales. Here we describe computational methods to reconstruct and navigate a multitude of high-resolution mSEM images of the human hip. We calculated cross-correlation shift vectors between overlapping images and used a mass-spring-damper model for optimal global registration...
November 2016: PLoS Computational Biology
https://www.readbyqxmd.com/read/27869911/controlled-production-of-sub-millimeter-liquid-core-hydrogel-capsules-for-parallelized-3d-cell-culture
#10
Hugo Doméjean, Mathieu de la Motte Saint Pierre, Anette Funfak, Nicolas Atrux-Tallau, Kevin Alessandri, Pierre Nassoy, Jérôme Bibette, Nicolas Bremond
Liquid core capsules having a hydrogel membrane are becoming a versatile tool for three-dimensional culture of micro-organisms and mammalian cells. Making sub-millimeter capsules at a high rate, via the breakup of a compound jet in air, opens the way to high-throughput screening applications. However, control of the capsule size monodispersity, especially required for quantitative bioassays, was still lacking. Here, we report how the understanding of the underlying hydrodynamic instabilities that occur during the process can lead to calibrated core-shell bioreactors...
December 20, 2016: Lab on a Chip
https://www.readbyqxmd.com/read/27627810/generation-of-an-in%C3%A2-vitro-3d-pdac-stroma-rich-spheroid-model
#11
Matthew J Ware, Vazrik Keshishian, Justin J Law, Jason C Ho, Carlos A Favela, Paul Rees, Billie Smith, Sayeeduddin Mohammad, Rosa F Hwang, Kimal Rajapakshe, Cristian Coarfa, Shixia Huang, Dean P Edwards, Stuart J Corr, Biana Godin, Steven A Curley
Pancreatic ductal adenocarcinoma (PDAC) is characterized by a prominent desmoplastic/stromal reaction, which contributes to the poor clinical outcome of this disease. Therefore, greater understanding of the stroma development and tumor-stroma interactions is highly required. Pancreatic stellate cells (PSC) are myofibroblast-like cells located in exocrine areas of the pancreas, which as a result of inflammation produced by PDAC migrate and accumulate in the tumor mass, secreting extracellular matrix components and producing the dense PDAC stroma...
November 2016: Biomaterials
https://www.readbyqxmd.com/read/27597161/discovery-of-a-new-structural-class-of-competitive-hdhodh-inhibitors-with-in-vitro-and-in-vivo-anti-inflammatory-immunosuppressive-effects
#12
Wenbin Shen, Xiao Ren, Jingtong Zhu, Yan Xu, Jie Lin, Yeying Li, Feng Zhao, Haizhou Zheng, Ruolan Li, Xiaolan Cui, Xuexia Zhang, Xinhua Lu, Zhihui Zheng
Human dihydroorotate dehydrogenase (hDHODH) is an inner mitochondrial membrane enzyme that involves in the fourth step of the biosynthesis of pyrimidine base. Inhibitors of hDHODH have been proven efficacy for the treatments of inflammation, rheumatoid arthritis, multiple sclerosis and cancer. In the present study, ascochlorin (ASC) and its derivatives, natural compounds from fungal metabolites, were discovered as hDHODH inhibitors by high-throughput screening. Enzyme kinetics studies showed that ASC competitively binds to hDHODH at the site of coenzyme Q substrate...
November 15, 2016: European Journal of Pharmacology
https://www.readbyqxmd.com/read/27564551/full-length-fibronectin-drives-fibroblast-accumulation-at-the-surface-of-collagen-microtissues-during-cell-induced-tissue-morphogenesis
#13
Jasper Foolen, Jau-Ye Shiu, Maria Mitsi, Yang Zhang, Christopher S Chen, Viola Vogel
Generating and maintaining gradients of cell density and extracellular matrix (ECM) components is a prerequisite for the development of functionality of healthy tissue. Therefore, gaining insights into the drivers of spatial organization of cells and the role of ECM during tissue morphogenesis is vital. In a 3D model system of tissue morphogenesis, a fibronectin-FRET sensor recently revealed the existence of two separate fibronectin populations with different conformations in microtissues, i.e. 'compact and adsorbed to collagen' versus 'extended and fibrillar' fibronectin that does not colocalize with the collagen scaffold...
2016: PloS One
https://www.readbyqxmd.com/read/27536780/redifferentiation-of-high-throughput-generated-fibrochondrocyte-micro-aggregates-impact-of-low-oxygen-tension
#14
Elke Berneel, Charlot Philips, Heidi Declercq, Ria Cornelissen
In meniscus tissue engineering strategies, enhancing the matrix quality of the neomeniscal tissue is important. When the differentiated phenotype of fibrochondrocytes is lost, the quality of the matrix becomes compromised. The objective of this study was to produce uniform fibrochondrocyte micro-aggregates with desirable phenotype and tissue homogeneity in large quantities using a simple and reproducible method. Furthermore, we investigated if hypoxia could enhance the matrix quality. Porcine fibrochondrocytes were expanded at 21% oxygen until passage 3 (P3) and a gene expression profile was determined...
2016: Cells, Tissues, Organs
https://www.readbyqxmd.com/read/27498178/combinatorial-extracellular-matrix-microenvironments-promote-survival-and-phenotype-of-human-induced-pluripotent-stem-cell-derived-endothelial-cells-in-hypoxia
#15
Luqia Hou, John Coller, Vanita Natu, Trevor J Hastie, Ngan F Huang
UNLABELLED: Recent developments in cell therapy using human induced pluripotent stem cell-derived endothelial cells (iPSC-ECs) hold great promise for treating ischemic cardiovascular tissues. However, poor post-transplantation viability largely limits the potential of stem cell therapy. Although the extracellular matrix (ECM) has become increasingly recognized as an important cell survival factor, conventional approaches primarily rely on single ECMs for in vivo co-delivery with cells, even though the endothelial basement membrane is comprised of a milieu of different ECMs...
October 15, 2016: Acta Biomaterialia
https://www.readbyqxmd.com/read/27481784/barcoding-the-largest-animals-on-earth-ongoing-challenges-and-molecular-solutions-in-the-taxonomic-identification-of-ancient-cetaceans
#16
REVIEW
Camilla Speller, Youri van den Hurk, Anne Charpentier, Ana Rodrigues, Armelle Gardeisen, Barbara Wilkens, Krista McGrath, Keri Rowsell, Luke Spindler, Matthew Collins, Michael Hofreiter
Over the last few centuries, many cetacean species have witnessed dramatic global declines due to industrial overharvesting and other anthropogenic influences, and thus are key targets for conservation. Whale bones recovered from archaeological and palaeontological contexts can provide essential baseline information on the past geographical distribution and abundance of species required for developing informed conservation policies. Here we review the challenges with identifying whale bones through traditional anatomical methods, as well as the opportunities provided by new molecular analyses...
September 5, 2016: Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences
https://www.readbyqxmd.com/read/27455635/shape-dependent-genotoxicity-of-mesoporous-silica-nanoparticles-and-cellular-mechanisms
#17
Meng Niu, Hongshan Zhong, Haibo Shao, Duo Hong, Tengchuang Ma, Ke Xu, Xiaowei Chen, Jinhang Han, Jun Sun
Although mesoporous silica nanoparticles (MSNs) are widely used in food products, cosmetics and nanomedicines as vector for drug delivery, data on their potential genotoxocity are limited. The aim of this study was to investigate the cytotoxic and genotoxic potentials of MSNs of different shapes, and to establish a high-throughput screening method for nanoparticles. We used functional macrophage receptor with collagenous structure (MARCO)-expressing DNA repair deficient chicken DT40 cells, which are designed to internalize nanoparticles and to be deficient in several specific DNA repair pathways...
March 2016: Journal of Nanoscience and Nanotechnology
https://www.readbyqxmd.com/read/27408951/high-throughput-collagen-fingerprinting-of-intact-microfaunal-remains-a-low-cost-method-for-distinguishing-between-murine-rodent-bones
#18
Mike Buckley, Muxin Gu, Sanu Shameer, Soyab Patel, Andrew T Chamberlain
RATIONALE: Microfaunal skeletal remains can be sensitive indicators of the contemporary ecosystem in which they are sampled and are often recovered in owl pellets in large numbers. Species identification of these remains can be obtained using a range of morphological criteria established for particular skeletal elements, but typically dominated by a reliance on cranial characters. However, this can induce biases under different environmental and taphonomic conditions. The aim of this research was to develop a high-throughput method of objectively identifying rodent remains from archaeological deposits using collagen fingerprinting, most notably the identification of rats from other myomorph rodents as a means to identify disturbances in the archaeofauna through the presence of invasive taxa not contemporary with the archaeological deposits...
April 15, 2016: Rapid Communications in Mass Spectrometry: RCM
https://www.readbyqxmd.com/read/27388239/umbilical-cord-derived-mesenchymal-stem-cell-derived-exosomal-micrornas-suppress-myofibroblast-differentiation-by-inhibiting-the-transforming-growth-factor-%C3%AE-smad2-pathway-during-wound-healing
#19
Shuo Fang, Chen Xu, Yuntong Zhang, Chunyu Xue, Chao Yang, Hongda Bi, Xijing Qian, Minjuan Wu, Kaihong Ji, Yunpeng Zhao, Yue Wang, Houqi Liu, Xin Xing
: : Excessive scar formation caused by myofibroblast aggregations is of great clinical importance during skin wound healing. Studies have shown that mesenchymal stem cells (MSCs) can promote skin regeneration, but whether MSCs contribute to scar formation remains undefined. We found that umbilical cord-derived MSCs (uMSCs) reduced scar formation and myofibroblast accumulation in a skin-defect mouse model. We found that these functions were mainly dependent on uMSC-derived exosomes (uMSC-Exos) and especially exosomal microRNAs...
October 2016: Stem Cells Translational Medicine
https://www.readbyqxmd.com/read/27321578/in-depth-proteomic-analysis-of-the-byssus-from-marine-mussel-mytilus-coruscus
#20
Chuan-Li Qin, Qi-Dong Pan, Qi Qi, Mei-Hua Fan, Jing-Jing Sun, Nan-Nan Li, Zhi Liao
UNLABELLED: Mussels attach to various submerged surfaces by using the byssus, which contains different proteins and is a promising source of water-resistant bio-adhesives for potential use in biotechnological and medical applications. The protein composition of the byssus has not yet been fully understood although at least eleven byssal proteins were characterized previously. In order to increase genomic resources and identify new byssal proteins from mussel Mytilus coruscus, high-throughput Illumina sequencing was undertaken on the foot, and 79,997,776 paired-ends reads were generated, yielding a library containing 88,825ft unigenes...
July 20, 2016: Journal of Proteomics
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