scholarly journals Imaging specific cellular glycan structures using glycosyltransferases via click chemistry

2017 ◽  
Author(s):  
Zhengliang L Wu ◽  
Anthony Person ◽  
Matthew Anderson ◽  
Barbara Burroughs ◽  
Timothy Tatge ◽  
...  
Keyword(s):  
Umbilical Vein ◽  
Functional Characterization ◽  
Tn Antigen ◽  
Stem Cell Line ◽  
Rate Limiting Step ◽  
Architectural Features ◽  
Huvec Cells ◽  
Umbilical Vein Endothelial Cells ◽  
Rate Limiting

AbstractHeparan sulfate (HS) is a polysaccharide fundamentally important for biologically activities. T/Tn antigens are universal carbohydrate cancer markers. Here, we report the specific imaging of these carbohydrates using a mesenchymal stem cell line and human umbilical vein endothelial cells (HUVEC). The staining specificities were demonstrated by comparing imaging of different glycans and validated by either removal of target glycans, which results in loss of signal, or installation of target glycans, which results in gain of signal. As controls, representative key glycans including O-GlcNAc, lactosaminyl glycans and hyaluronan were also imaged. HS staining revealed novel architectural features of the extracellular matrix (ECM) of HUVEC cells. Results from T/Tn antigen staining suggest that O-GalNAcylation is a rate-limiting step for O-glycan synthesis. Overall, these highly specific approaches for HS and T/Tn antigen imaging should greatly facilitate the detection and functional characterization of these biologically important glycans.

scholarly journals Hexose transport in Torulaspora delbrueckii: identification of Igt1, a new dual-affinity transporter

2020 ◽  
Vol 20 (1) ◽  
Author(s):  
Andreia Pacheco ◽  
Lorena Donzella ◽  
Maria Jose Hernandez-Lopez ◽  
Maria Judite Almeida ◽  
Jose Antonio Prieto ◽  
...  
Keyword(s):  
Glucose Transporter ◽  
Functional Characterization ◽  
Sugar Transport ◽  
Sugar Metabolism ◽  
Torulaspora Delbrueckii ◽  
Sugar Transporters ◽  
Rate Limiting Step ◽  
Efficient Fermentation ◽  
Rate Limiting

ABSTRACT Torulaspora delbrueckii is a yeast species receiving increasing attention from the biotechnology industry, with particular relevance in the wine, beer and baking sectors. However, little is known about its sugar transporters and sugar transport capacity, frequently a rate-limiting step of sugar metabolism and efficient fermentation. Actually, only one glucose transporter, Lgt1, has been characterized so far. Here we report the identification and characterization of a second glucose transporter gene, IGT1, located in a cluster, upstream of LGT1 and downstream of two other putative hexose transporters. Functional characterization of IGT1 in a Saccharomyces cerevisiae hxt-null strain revealed that it encodes a transporter able to mediate uptake of glucose, fructose and mannose and established that its affinity, as measured by Km, could be modulated by glucose concentration in the medium. In fact, IGT1-transformed S. cerevisiae hxt-null cells, grown in 0.1% glucose displayed biphasic glucose uptake kinetics with an intermediate- (Km = 6.5 ± 2.0 mM) and a high-affinity (Km = 0.10 ± 0.01 mM) component, whereas cells grown in 2% glucose displayed monophasic kinetics with an intermediate-affinity (Km of 11.5 ± 1.5 mM). This work contributes to a better characterization of glucose transport in T. delbrueckii, with relevant implications for its exploitation in the food industry.

Partial Characterization of a Fibrinolytic Inhibitor Produced by Cultured Endothelial Cells Derived from Human Umbilical Vein

1982 ◽  
Vol 47 (02) ◽  
pp. 128-131 ◽  
Author(s):  
F Esnard ◽  
E Dupuy ◽  
A M Dosne ◽  
E Bodevin
Keyword(s):  
Endothelial Cells ◽  
Primary Culture ◽  
Ammonium Sulphate ◽  
Concanavalin A ◽  
Umbilical Vein ◽  
Human Umbilical Vein ◽  
Preliminary Characterization ◽  
Umbilical Vein Endothelial Cells ◽  
Ammonium Sulphate Saturation

SummaryA preliminary characterization of a fibrinolytic inhibitor released by human umbilical vein endothelial cells in primary culture is reported. This molecule of Mr comprised between 2 × 105 and 106 and of μ2 mobility precipitates at 43% ammonium sulphate saturation and is totally adsorbed on Concanavalin A Sepharose 4 B. A possible relationship with a macroglobulins is discussed.

scholarly journals Photodynamic therapy targeting VCAM-1-expressing human umbilical vein endothelial cells using a PpIX–VCAM-1 binding peptide–quantum dot conjugate

RSC Advances ◽  
2017 ◽  
Vol 7 (80) ◽  
pp. 50562-50570 ◽  
Author(s):  
Huijuan Yin ◽  
Xiafei Shi ◽  
Hong Wang ◽  
Wendong Jin ◽  
Yingxin Li ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Photodynamic Therapy ◽  
Quantum Dot ◽  
Umbilical Vein ◽  
Photodynamic Effect ◽  
Binding Peptide ◽  
Human Umbilical Vein ◽  
Huvec Cells ◽  
Umbilical Vein Endothelial Cells

Enhanced PDT was induced by the conjugate of PpIX (photodynamic effect)–VCAM-1 binding peptide (target)–QD (carrier) by the augmented ROS on VCAM-1 expressing HUVEC cells.

Production and Functional Characterization of Two Mouse/Human Chimeric Antibodies With Specificity for the Tumor-Associated Tn-Antigen

Hybridoma ◽  
2000 ◽  
Vol 19 (3) ◽  
pp. 229-239 ◽  
Author(s):  
Pablo Oppezzo ◽  
Eduardo Osinaga ◽  
Diana Tello ◽  
Sylvie Bay ◽  
Daniele Cantacuzene ◽  
...  
Keyword(s):  
Functional Characterization ◽  
Tn Antigen ◽  
Chimeric Antibodies

scholarly journals Characterization of the interactions between procoagulant albumin and human endothelial cells

Blood ◽  
1993 ◽  
Vol 82 (9) ◽  
pp. 2684-2692
Author(s):  
KJ Faucette ◽  
LA Fitzgerald ◽  
L Liu ◽  
CJ Parker ◽  
GM Rodgers
Keyword(s):  
Endothelial Cells ◽  
Umbilical Vein ◽  
Quantitative Polymerase Chain Reaction ◽  
Scavenger Receptor ◽  
Kinase C ◽  
Normal Human ◽  
Membrane Bound ◽  
Polymerase Chain ◽  
Umbilical Vein Endothelial Cells

Normal human plasma contains procoagulant albumin (PC-Al), an anionic form of albumin that induces tissue factor (TF) activity in human umbilical vein endothelial cells (HUVEC) and monocytes. In this study, we investigated both the interactions between HUVEC and PC-Al and the mechanism by which PC-Al induces TF activity. Binding of PC-Al to HUVEC was specific and reversible. Further studies indicated that membrane- bound PC-Al was not internalized by HUVEC. A potential receptor on HUVEC was suggested by studies in which the capacity of a variety of reagents to inhibit the activity of PC-Al was quantitated. Induction of TF activity by PC-Al was antagonized by dextran sulfate, heparin, fucoidan, and concanavalin A but not by ovalbumin, polyglutamic acid, or polyvinyl sulfate. This competition profile bears similarities to those reported for scavenger receptors that have been identified on both HUVEC and monocytes. Involvement of protein kinase C (PKC) in the PC-Al-induced enhancement of TF activity was suggested by experiments in which staurosporine, an inhibitor of PKC, suppressed the activity of PC-Al. The induction of TF activity by PC-Al was further characterized by using a quantitative polymerase chain reaction assay. Increased TF mRNA was first seen after 1 hour of incubation with PC-Al. Maximal observed expression occurred at 2 hours, but at 5 hours, expression had significantly decreased. Monocytes could also be induced to express TF mRNA after a 2-hour incubation with PC-Al. These results suggest that the functionally relevant binding of PC-Al to HUVEC may be mediated through interactions with a membrane constituent that has some of the properties of a scavenger receptor and that this interaction augments TF activity by enhancing transcription of TF mRNA, at least in part, by a mechanism that is dependent on activation of PKC.

scholarly journals Parallel Isolation and Characterization of Porcine Smooth Muscle, Endothelial and Mesenchymal Stromal Cells for Bioengineering Applications

2021 ◽  
Author(s):  
Sara Morini ◽  
Iris Pla-Palacín ◽  
Pilar Sainz-Arnal ◽  
Natalia Sánchez-Romero ◽  
Maria Falceto ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Cultured Cells ◽  
Umbilical Vein ◽  
Building Blocks ◽  
Cell Types ◽  
Aortic Smooth Muscle ◽  
Isolation And Characterization ◽  
Umbilical Vein Endothelial Cells

Abstract There is significant interest in the pig as the animal model of choice for organ transplantation and the study of tissue engineering (TE) products and applications. Currently, efforts are being taken to bioengineer solid organs to reduce donor shortages for transplantation. For complex organs such as the lung, heart, and liver, the vasculature represents a fundamental feature. Thus, to generate organs with a functional vascular network, the different cells constituting the building blocks of the blood vessels should be procured. However, due to species' specificities, porcine cell isolation, expansion, and characterization are not entirely straightforward compared to human cell procurement. Here, we report the establishment of simple and suitable methods for the isolation and characterization of distinct porcine cells for bioengineering purposes.We successfully isolated, expanded and characterized porcine bone marrow-derived mesenchymal stromal (pBM-MSC), aortic smooth muscle (pASMC), and umbilical vein endothelial cells (pUVEC). We demonstrated that the three cell types showed specific immunophenotypical features. Moreover, we demonstrated that pBM-MSC could preserve their multipotency in vitro, and pUVEC were capable of maintaining their functionality in vitro.These cultured cells could be further expanded and represent a useful cellular tool for TE purposes (i.e., for recellularization approaches of vascularized organs or in vitro angiogenesis studies).

Abstract 5078: Physical Training Reduces the Ability of Serum from Patients with Heart Failure to Induce Endothelial Cell Death

Circulation ◽  
2008 ◽  
Vol 118 (suppl_18) ◽  
Author(s):  
Caterina Mammi ◽  
Massimiliano Caprio ◽  
Stefania Gorini ◽  
Andrea la Sala ◽  
Antonella Antelmi ◽  
...  
Keyword(s):  
Heart Failure ◽  
Endothelial Cells ◽  
Cell Death ◽  
Cardiac Rehabilitation ◽  
Physical Training ◽  
Umbilical Vein ◽  
Apoptotic Pathway ◽  
Patients With Heart Failure ◽  
Huvec Cells ◽  
Umbilical Vein Endothelial Cells

Physical training improves endothelial function and exercise capacity in patients with heart failure (HF). Serum from patients with cardiovascular diseases shows increased ability to induce apoptosis in human endothelial cells potentially contributing to the progression of atherosclerotic and endothelial dysfunction. Aim of this study was to evaluate the effect of cardiac rehabilitation on the pro-apoptotic properties of serum in patients with heart failure. Human Umbilical Vein Endothelial Cells (HUVECs) were incubated with 20% serum from 24 patients with HF (HYHA II) before and after cardiovascular rehabilitation. For apoptosis quantification, fragmented DNA was determined using Cell Death detection ELISA kit and propidium iodide (PI) staining analyzed by flow cytometry. The expression on endothelial cells of proteins involved in apoptotic pathway (bax, caspasi-3, bcl-2) was studied by Western blot analysis. Necrosis was evaluated measuring the lactate dehydrogenase released from damaged cells into the supernatant using a cytotoxicity detection kit. Serum from patients with heart failure showed a significantly greater ability to induce apoptosis (67 ± 4,6% vs 23 ± 5,2%, p<0,001) and necrosis (44 ± 3% vs 32 ± 2,1%, p<0,05) of HUVEC cells compared with controls. Cardiac rehabilitation significantly reduced the ability of patient serum to induce apoptosis (PI: 40.44% ± 3,9 vs 25.22% ± 2,6, p<0,01; ELISA: 67 ± 4,6% vs 42 ± 4,4%, p<0,001) and necrosis (44 ± 3% vs 33 ± 2%, p<0,01) of HUVEC cells compared to baseline. Moreover endothelial cells treated with serum after rehabilitation showed a significant decrease of bax expression (1,12 ± 0,06 vs 0,89 ± 0,08, p<0,05) and a significant decrease of cleaved Caspase-3 expression (0,91 ± 0,16 vs 0,72 ± 0,08, p<0.05). Serum of patients with heart failure has increased pro-apoptotic properties. Physical training reduces the ability of serum-contained factors to induce cell death of endothelial cells in patient with heart failure.

Wavelet Based Structural Analysis of Electroless Deposits in the Diffusion Limited Regime

1994 ◽  
Vol 367 ◽  
Author(s):  
A. Arneodo ◽  
F. Argoul ◽  
A. Kuhn ◽  
J.F. Muzy
Keyword(s):  
Electroless Deposition ◽  
Side Reactions ◽  
Statistical Features ◽  
Diffusion Limited ◽  
Rate Limiting Step ◽  
Limiting Step ◽  
Fractal Patterns ◽  
Rate Limiting ◽  
Comparative Structural Characterization

AbstractWe discuss the actual relevance of thin gap geometry electrodeposition to generate fractal patterns that mimic the morphology of Witten and Sander's diffusion-limited aggregates (DLA). Eliminating migration and convection, as well as electrochemical side reactions, we show that electroless deposition is a good candidate to meet the requirements for diffusion to be the rate limiting step of the growth process. We use the wavelet transform microscope to achieve a comparative structural characterization of both experimental electroless deposits and numerical DLA clusters. The fact that five-fold symmetry and Fibonacci hierarchical ordering are found as common predominant statistical features is, to our knowledge, the first demonstration, relying on an appropriate structural fractal analysis, of the existence of DLA morphologies in an experimental context.

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