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https://www.readbyqxmd.com/read/28595551/nmr-based-drug-development-and-improvement-against-malignant-melanoma-implications-for-the-mia-protein-family
#1
Oliver Arnolds, Xueyin Zhong, King Tuo Yip, Miriam Schöpel, Bastian Kohl, Stefanie Pütz, Raid Abdel-Jalil, Raphael Stoll
The Melanoma Inhibitory Activity (MIA) protein is strongly expressed and secreted by malignant melanoma cells and was shown to promote melanoma development and invasion. The MIA protein was the first extracellular protein shown to adopt a Src homology 3 (SH3) domain-like fold in solution that can bind to fibronectin type III domains. Together with MIA, the homologous proteins OTOR (or FDP), MIA-2, and TANGO (or MIA-3) constitute a protein family of non-cytoplasmic and - except for full-length TANGO and TANGO1-like (TALI) - extracellular SH3-domain containing proteins...
June 8, 2017: Current Medicinal Chemistry
https://www.readbyqxmd.com/read/28559982/long-noncoding-rnas-and-novel-inflammatory-genes-determined-by-rna-sequencing-in-human-lymphocytes-are-up-regulated-in-permanent-atrial-fibrillation
#2
Xue-Jing Yu, Li-Hui Zou, Jun-Hua Jin, Fei Xiao, Li Li, Nan Liu, Jie-Fu Yang, Tong Zou
Atrial fibrillation (AF) is a common arrhythmia in clinical practice. Currently, approximately 33.5 million individuals are affected by AF globally. AF involves multiple complicated mechanisms which have not been fully investigated yet. RNA sequencing (RNAseq) is an outstanding method for investigation of diseases due to its high-throughput information. Here, RNAseq was applied to determine mRNA and long noncoding RNA (lncRNA) expression profiles in human lymphocytes from 6 permanent atrial fibrillation (pmAF) patients and 6 healthy controls...
2017: American Journal of Translational Research
https://www.readbyqxmd.com/read/28455445/the-soluble-protease-adamdec1-released-from-activated-platelets-hydrolyzes-platelet-membrane-pro-epidermal-growth-factor-egf-to-active-high-molecular-weight-egf
#3
Rui Chen, Ge Jin, Thomas M McIntyre
Platelets are the sole source of EGF in circulation, yet how EGF is stored or released from stimulated cells is undefined. In fact, we found platelets did not store EGF, synthesized as a single 6-kDa domain in pro-EGF, but rather expressed intact pro-EGF precursor on granular and plasma membranes. Activated platelets released high-molecular-weight (HMW)-EGF, produced by a single cleavage between the EGF and the transmembrane domains of pro-EGF. We synthesized a fluorogenic peptide encompassing residues surrounding the putative sessile arginyl residue and found stimulated platelets released soluble activity that cleaved this pro-EGF1020-1027 peptide...
June 16, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28448052/an-in-vitro-organ-culture-model-of-the-murine-intervertebral-disc
#4
Jennifer W Liu, Kevin H Lin, Christian Weber, Sameer Bhalla, Sean Kelso, Kaixi Wang, Simon Y Tang
Intervertebral disc (IVD) degeneration is a significant contributor to low back pain. The IVD is a fibrocartilaginous joint that serves to transmit and dampen loads in the spine. The IVD consists of a proteoglycan-rich nucleus pulposus (NP) and a collagen-rich annulus fibrosis (AF) sandwiched by cartilaginous end-plates. Together with the adjacent vertebrae, the vertebrae-IVD structure forms a functional spine unit (FSU). These microstructures contain unique cell types as well as unique extracellular matrices...
April 11, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28444343/lysolecithin-as-feed-additive-enhances-collagen-expression-and-villus-length-in-the-jejunum-of-broiler-chickens
#5
D L Brautigan, R Li, E Kubicka, S D Turner, J S Garcia, M L Weintraut, E A Wong
Adding lysolecithin to feed has reportedly improved the performance of broiler chickens. Lysolecithin is generated by phospholipase catalyzed hydrolysis of lecithin. The enzymatic reaction converts various phospholipids into the corresponding lysophospholipids, with lysophosphatidylcholine (LPC) one of the primary products. Here we compared supplementation with a commercial lysolecithin (Lysoforte®) with comparable levels of highly purified LPC for effects on broilers. Despite no differences in weight gain during the starter period, we discovered a significant increase in average villus length with lysolecithin and an increase in villus width with purified LPC...
April 24, 2017: Poultry Science
https://www.readbyqxmd.com/read/28406494/laser-capture-microdissection-of-pancreatic-acinar-cells-to-identify-proteomic-alterations-in-a-murine-model-of-caerulein-induced-pancreatitis
#6
John P Shapiro, Hannah M Komar, Baris Hancioglu, Lianbo Yu, Ming Jin, Yuko Ogata, Phil A Hart, Zobeida Cruz-Monserrate, Gregory B Lesinski, Darwin L Conwell
OBJECTIVES: Chronic pancreatitis (CP) is characterized by inflammation and fibrosis of the pancreas, leading to pain, parenchymal damage, and loss of exocrine and endocrine function. There are currently no curative therapies; diagnosis remains difficult and aspects of pathogenesis remain unclear. Thus, there is a need to identify novel biomarkers to improve diagnosis and understand pathophysiology. We hypothesize that pancreatic acinar regions contain proteomic signatures relevant to disease processes, including secreted proteins that could be detected in biofluids...
April 13, 2017: Clinical and Translational Gastroenterology
https://www.readbyqxmd.com/read/28393762/engineering-a-functional-three-dimensional-human-cardiac-tissue-model-for-drug-toxicity-screening
#7
Hong Fang Lu, Meng Fatt Leong, Tze Chiun Lim, Ying Ping Chua, Jia Kai Lim, Chan Du, Andrew C A Wan
Cardiotoxicity is one of the major reasons for clinical drug attrition. In vitro tissue models that can provide efficient and accurate drug toxicity screening are highly desired for preclinical drug development and personalized therapy. Here, we report the fabrication and characterization of a human cardiac tissue model for high throughput drug toxicity studies. Cardiac tissues were fabricated via cellular self-assembly of human transgene-free induced pluripotent stem cells-derived cardiomyocytes in pre-fabricated polydimethylsiloxane molds...
May 11, 2017: Biofabrication
https://www.readbyqxmd.com/read/28355300/evaluation-of-synthetic-vascular-grafts-in-a-mouse-carotid-grafting-model
#8
Alex H P Chan, Richard P Tan, Praveesuda L Michael, Bob S L Lee, Laura Z Vanags, Martin K C Ng, Christina A Bursill, Steven G Wise
Current animal models for the evaluation of synthetic grafts are lacking many of the molecular tools and transgenic studies available to other branches of biology. A mouse model of vascular grafting would allow for the study of molecular mechanisms of graft failure, including in the context of clinically relevant disease states. In this study, we comprehensively characterise a sutureless grafting model which facilitates the evaluation of synthetic grafts in the mouse carotid artery. Using conduits electrospun from polycaprolactone (PCL) we show the gradual development of a significant neointima within 28 days, found to be greatest at the anastomoses...
2017: PloS One
https://www.readbyqxmd.com/read/28337850/high-throughput-screening-of-vascular-endothelium-destructive-or-protective-microenvironments-cooperative-actions-of-extracellular-matrix-composition-stiffness-and-structure
#9
Yonghui Ding, Michael Floren, Wei Tan
Pathological modification of the subendothelial extracellular matrix (ECM) has closely been associated with endothelial activation and subsequent cardiovascular disease progression. To understand regulatory mechanisms of these matrix modifications, the majority of previous efforts have focused on the modulation of either chemical composition or matrix stiffness on 2D smooth surfaces without simultaneously probing their cooperative effects on endothelium function on in vivo like 3D fibrous matrices. To this end, a high-throughput, combinatorial microarray platform on 2D and 3D hydrogel settings to resemble the compositions, stiffness, and structure of healthy and diseased subendothelial ECM has been established, and further their respective and combined effects on endothelial attachment, proliferation, inflammation, and junctional integrity have been investigated...
June 2017: Advanced Healthcare Materials
https://www.readbyqxmd.com/read/28322869/scaffold-free-high-throughput-generation-of-quiescent-valvular-microtissues
#10
Annelies Roosens, Inès Puype, Ria Cornelissen
AIMS: The major challenge of working with valvular interstitial cells in vitro is the preservation or recovery of their native quiescent state. In this study, a biomimetic approach is used which aims to engineer small volume, high quality valve microtissues, having a potential in regenerative medicine and as a relevant 3D in vitro model to provide insights into valve (patho)biology. METHODS AND RESULTS: To form micro-aggregates, porcine valvular interstitial cells were seeded in agarose micro-wells and cultured in medium supplemented with 250μM Ascorbic Acid 2-phosphate for 22days...
March 18, 2017: Journal of Molecular and Cellular Cardiology
https://www.readbyqxmd.com/read/28298644/the-combination-of-three-dimensional-and-rotary-cell-culture-system-promotes-the-proliferation-and-maintains-the-differentiation-potential-of-rat-bmscs
#11
Yilong Tang, Yan Xu, Zhifeng Xiao, Yannan Zhao, Jing Li, Sufang Han, Lei Chen, Bin Dai, Ling Wang, Bing Chen, Hong Wang
Bone marrow mesenchymal stem cells (BMSCs) are a good candidate for tissue engineering and clinical application. One of the challenges in its cell therapy is how to quickly obtain an adequate number of seed cells and meanwhile maintain suitable differentiation potential. In this study we combined three-dimensional (3D) collagen porous scaffolds with rotary cell culture system (RCCS) (RCCS-3D) to create a stereoscopic dynamic environment for the amplification of rat BMSCs in vitro. The results revealed that this RCCS-3D system could enhance BMSCs' proliferation and colony formation, as well as maintain the differentiation potential compared with conventional static two-dimensional (2D) and 3D cell culture conditions...
March 15, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28255014/macrothrombocytopenia-and-dense-granule-deficiency-associated-with-fli1-variants-ultrastructural-and-pathogenic-features
#12
Paul Saultier, Léa Vidal, Matthias Canault, Denis Bernot, Céline Falaise, Catherine Pouymayou, Jean-Claude Bordet, Noémie Saut, Agathe Rostan, Véronique Baccini, Franck Peiretti, Marie Favier, Pauline Lucca, Jean-François Deleuze, Robert Olaso, Anne Boland, Pierre Emmanuel Morange, Christian Gachet, Fabrice Malergue, Sixtine Fauré, Anita Eckly, David-Alexandre Trégouët, Marjorie Poggi, Marie-Christine Alessi
Congenital macrothrombocytopenia is a family of rare diseases, of which a significant fraction remains to be genetically characterized. To analyze cases of unexplained thrombocytopenia, 27 individuals from a patient cohort of the Bleeding and Thrombosis Exploration Center of the University Hospital of Marseille were recruited for a high-throughput gene sequencing study. This strategy led to the identification of two novel FLI1 variants (c.1010G>A and c.1033A>G) responsible for macrothrombocytopenia. The FLI1 variant carriers' platelets exhibited a defect in aggregation induced by low-dose adenosine diphosphate (ADP), collagen and thrombin receptor-activating peptide (TRAP), a defect in adenosine triphosphate (ATP) secretion, a reduced mepacrine uptake and release and a reduced CD63 expression upon TRAP stimulation...
June 2017: Haematologica
https://www.readbyqxmd.com/read/28240272/a-microfabricated-96-well-3d-assay-enabling-high-throughput-quantification-of-cellular-invasion-capabilities
#13
Rui Hao, Yuanchen Wei, Chaobo Li, Feng Chen, Deyong Chen, Xiaoting Zhao, Shaoliang Luan, Beiyuan Fan, Wei Guo, Junbo Wang, Jian Chen
This paper presents a 96-well microfabricated assay to study three-dimensional (3D) invasion of tumor cells. A 3D cluster of tumor cells was first generated within each well by seeding cells onto a micro-patterned surface consisting of a central fibronectin-coated area that promotes cellular attachment, surrounded by a poly ethylene glycol (PEG) coated area that is resistant to cellular attachment. Following the formation of the 3D cell clusters, a 3D collagen extracellular matrix was formed in each well by thermal-triggered gelation...
February 27, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28216326/a-scalable-lysyl-hydroxylase-2-expression-system-and-luciferase-based-enzymatic-activity-assay
#14
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...
March 15, 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
#15
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
#16
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
#17
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...
April 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
#18
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
#19
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
#20
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
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