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https://www.readbyqxmd.com/read/28205171/synthesis-and-live-cell-imaging-of-fluorescent-sterols-for-analysis-of-intracellular-cholesterol-transport
#1
Maciej Modzel, Frederik W Lund, Daniel Wüstner
Cellular cholesterol homeostasis relies on precise control of the sterol content of organelle membranes. Obtaining insight into cholesterol trafficking pathways and kinetics by live-cell imaging relies on two conditions. First, one needs to develop suitable analogs that resemble cholesterol as closely as possible with respect to their biophysical and biochemical properties. Second, the cholesterol analogs should have good fluorescence properties. This interferes, however, often with the first requirement, such that the imaging instrumentation must be optimized to collect photons from suboptimal fluorophores, but good cholesterol mimics, such as the intrinsically fluorescent sterols, cholestatrienol (CTL) or dehydroergosterol (DHE)...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28204818/reactive-oxygen-species-mediate-angiotensin-ii-induced-transcytosis-of-low-density-lipoprotein-across-endothelial-cells
#2
Fang Bian, Jun Cui, Tao Zheng, Si Jin
The retention of plasma low-density lipoprotein (LDL) particles to subendothelial spaces through transcytosis across the endothelium is the initial step of atherosclerosis (AS). Angiotensin II (Ang II), as the principal effector molecule of the renin-angiotensin system (RAS), is implicated in several important steps of AS development. However, whether or not Ang II can directly exert a pro‑atherogenic effect by promoting LDL transcytosis across endothelial barriers, has not been defined. In the present study, we found that Ang II upregulated intracellular reactive oxygen species (ROS) levels in endothelial cells (ECs) by measuring fluorescence of 2',7'-dichlorofluorescein (DCF‑DA)...
February 13, 2017: International Journal of Molecular Medicine
https://www.readbyqxmd.com/read/28194311/tracking-the-virus-like-particles-of-macrobrachium-rosenbergii-nodavirus-in-insect-cells
#3
Ummi Fairuz Hanapi, Chean Yeah Yong, Zee Hong Goh, Noorjahan Banu Alitheen, Swee Keong Yeap, Wen Siang Tan
Macrobrachium rosenbergii nodavirus (MrNv) poses a major threat to the prawn industry. Currently, no effective vaccine and treatment are available to prevent the spread of MrNv. Its infection mechanism and localisation in a host cell are also not well characterised. The MrNv capsid protein (MrNvc) produced in Escherichia coli self-assembled into virus-like particles (VLPs) resembling the native virus. Thus, fluorescein labelled MrNvc VLPs were employed as a model to study the virus entry and localisation in Spodoptera frugiperda, Sf9 cells...
2017: PeerJ
https://www.readbyqxmd.com/read/28188926/redox-modification-of-caveolar-proteins-in-the-cardiovascular-system-role-in-cellular-signalling-and-disease
#4
REVIEW
Kristen J Bubb, Asa Birna Birgisdottir, Owen Tang, Thomas Hansen, Gemma A Figtree
Rapid and coordinated release of a variety of reactive oxygen species (ROS) such as superoxide (O2(.-)), hydrogen peroxide (H2O2) and peroxynitrite, in specific microdomains, play a crucial role in cell signalling in the cardiovascular system. These reactions are mediated by reversible and functional modifications of a wide variety of key proteins. Dysregulation of this oxidative signaling occurs in almost all forms of cardiovascular disease (CVD), including at the very early phases. Despite the heavily publicized failure of "antioxidants" to improve CVD progression, pharmacotherapies such as those targeting the renin-angiotensin system, or statins, exert at least part of their large clinical benefit via modulating cellular redox signalling...
February 7, 2017: Free Radical Biology & Medicine
https://www.readbyqxmd.com/read/28187162/loss-of-caveolin-1-alters-extracellular-matrix-protein-expression-and-ductal-architecture-in-murine-mammary-glands
#5
Christopher Thompson, Sahar Rahim, Jeremiah Arnold, Abigail Hielscher
The extracellular matrix (ECM) is abnormal in breast tumors and has been reported to contribute to breast tumor progression. One factor, which may drive ongoing matrix synthesis in breast tumors, is the loss of stromal caveolin-1 (cav-1), a scaffolding protein of caveolae, which has been linked to breast tumor aggressiveness. To determine whether loss of cav-1 results in the abnormal expression of matrix proteins, mammary glands from cav- 1-/- and cav- 1 +/+ mice were investigated for differences in expression of several ECM proteins...
2017: PloS One
https://www.readbyqxmd.com/read/28186394/mechanisms-of-ph-sensitivity-and-cellular-internalization-of-peoz-b-pla-micelles-with-varied-hydrophilic-hydrophobic-ratios-and-intracellular-trafficking-routes-and-fate-of-the-copolymer
#6
Dishi Wang, Yanxia Zhou, Xinru Li, Xiaoyou Qu, Yunqiang Deng, Ziqi Wang, Chuyu He, Yang Zou, Yiguang Jin, Yan Liu
pH-responsive polymeric micelles have shown promise for the targeted and intracellular delivery of antitumor agents. The present study aimed to elucidate the possible mechanisms of pH-sensitivity and cellular internalization of PEOz-b-PLA micelles in detail, further unravel the effect of hydrophilic/hydrophobic ratio of the micelles on their cellular internalization, and examine the intracellular trafficking routes and fate of PEOz-b-PLA after internalization of the micelles. The results of variations in the size and Zeta potential of PEOz-b-PLA micelles and cross-sectional area of PEOz-b-PLA molecules with pH values suggested that electrostatic repulsion between PEOz chains resulting from ionization of the tertiary amide groups along PEOz chain at lower pH than its pKa was responsible for pH-sensitivity of PEOz-b-PLA micelles...
February 10, 2017: ACS Applied Materials & Interfaces
https://www.readbyqxmd.com/read/28184336/versatile-functions-of-caveolin-1-in-aging-related-diseases
#7
REVIEW
Kim Cuc Thi Nguyen, Kyung A Cho
Caveolin-1 (Cav-1) is a trans-membrane protein that is a major component of the caveolae structure on the plasma membrane. Cav-1 is involved in the regulation of various cellular processes, including cell growth, differentiation, endocytosis, and in particular it has been implied in cellular senescence. Here we review current knowledge about Cav-1 in cellular signaling and discuss the role of Cav-1 in aging-related diseases.
January 2017: Chonnam Medical Journal
https://www.readbyqxmd.com/read/28181394/effects-of-silver-nanoparticles-on-the-interactions-of-neuron-and-glia-like-cells-toxicity-uptake-mechanisms-and-lysosomal-tracking
#8
I-Lun Hsiao, Yi-Kong Hsieh, Chun-Yu Chuang, Chu-Fang Wang, Yuh-Jeen Huang
Silver nanoparticles (AgNPs) are commonly used nanomaterials in consumer products. Previous studies focused on its effects on neurons; however, little is known about their effects and uptake mechanisms on glial cells under normal or activated states. Here, ALT astrocyte-like, BV-2 microglia and differentiated N2a neuroblastoma cells were directly or indirectly exposed to 10 nm AgNPs using mono- and co-culture system. A lipopolysaccharide (LPS) was pretreated to activate glial cells before AgNP treatment for mimicking NP exposure under brain inflammation...
February 9, 2017: Environmental Toxicology
https://www.readbyqxmd.com/read/28179028/betanodavirus-like-particles-enter-host-cells-via-clathrin-mediated-endocytosis-in-a-cholesterol-ph-and-cytoskeleton-dependent-manner
#9
Runqing Huang, Guohua Zhu, Jing Zhang, Yuxiong Lai, Yu Xu, Jianguo He, Junfeng Xie
Betanodavirus, also referred to nervous necrosis virus (NNV), is the causative agent of the fatal disease, viral nervous necrosis and has brought significant economic losses in marine and freshwater cultured fish, especially larvae and juveniles. Here, we used an established invasion model with virus-like particle (VLP)-cells, mimicking orange-spotted grouper nervous necrosis virus (OGNNV), to investigate the crucial events of virus entry. VLP were observed in the perinuclear regions of Asian sea bass (SB) cells within 1...
February 8, 2017: Veterinary Research
https://www.readbyqxmd.com/read/28167128/testosterone-promotes-tube-formation-of-endothelial-cells-isolated-from-veins-via-activation-of-smad1-protein
#10
Pei Liu, Xiaosa Li, Fuhu Song, Ping Li, Jinzhi Wei, Qing Yan, Xingyan Xu, Jun Yang, Chuanxiang Li, Xiaodong Fu
Testosterone (T) deficiency is positively correlated with the increased incidence of cardiovascular disease. However, the effects of T on vascular endothelial cells remain obscure. Tube formation capacity is critical for vascular regeneration/repair and Smad1 plays an important role in these events. In this study, we investigated the effects of T on Smad1 activation and tube formation of cultured human umbilical endothelial cells (HUVECs). Our results showed that T rapidly increased endothelial Smad1 phosphorylation...
February 3, 2017: Molecular and Cellular Endocrinology
https://www.readbyqxmd.com/read/28161538/chloride-channel-clc-2-enhances-intestinal-epithelial-tight-junction-barrier-function-via-regulation-of-caveolin-1-and-caveolar-trafficking-of-occludin
#11
Prashant K Nighot, Lana Leung, Thomas Y Ma
Previous studies have demonstrated that the chloride channel ClC-2 plays a critical role in intestinal epithelial tight junction (TJ) barrier function via intracellular trafficking of TJ protein occludin. To study the mechanism of ClC-2-mediated TJ barrier function and intracellular trafficking of occludin, we established ClC-2 over-expressing Caco-2 cell line (Caco-2(CLCN2)) by full length ClC-2 ORF transfection. ClC-2 over-expression (Caco-2(CLCN2)) significantly enhanced TJ barrier (increased TER by ≥2 times and reduced inulin flux by 50%) compared to control Caco-2(pEZ) cells...
February 2, 2017: Experimental Cell Research
https://www.readbyqxmd.com/read/28160133/cardioprotective-kinase-signaling-to-subsarcolemmal-and-interfibrillar-mitochondria-is-mediated-by-caveolar-structures
#12
Wylly Ramsés García-Niño, Francisco Correa, Julia Isabel Rodríguez-Barrena, Juan Carlos León-Contreras, Mabel Buelna-Chontal, Elizabeth Soria-Castro, Rogelio Hernández-Pando, José Pedraza-Chaverri, Cecilia Zazueta
The demonstration that caveolin-3 overexpression reduces myocardial ischemia/reperfusion injury and our own finding that multiprotein signaling complexes increase in mitochondria in association with caveolin-3 levels, led us to investigate the contribution of caveolae-driven extracellular signal-regulated kinases 1/2 (ERK1/2) on maintaining the function of cardiac mitochondrial subpopulations from reperfused hearts subjected to postconditioning (PostC). Rat hearts were isolated and subjected to ischemia/reperfusion and to PostC...
March 2017: Basic Research in Cardiology
https://www.readbyqxmd.com/read/28154158/caveolae-provide-a-specialized-membrane-environment-for-respiratory-syncytial-virus-assembly
#13
Alexander Ludwig, Tra Huong Nguyen, Daniel Leong, Laxmi Iyer Ravi, Tan Boon Huan, Sara Sandin, Richard J Sugrue
Respiratory syncytial virus (RSV) is an enveloped virus that assembles into filamentous virus particles on the surface of infected cells. Morphogenesis of RSV is dependent upon cholesterol-rich (lipid raft) membrane microdomains, but the specific role of individual raft molecules in RSV assembly is not well defined. Here we show that RSV morphogenesis occurs within caveolar membranes and that both caveolin-1 and cavin-1, the two major structural and functional components of caveolae, are actively recruited to and incorporated into the RSV envelope...
February 2, 2017: Journal of Cell Science
https://www.readbyqxmd.com/read/28149711/hepatic-caveolin-1-is-enhanced-in-cyp27a1-apoe-double-knockout-mice
#14
Line Zurkinden, Yosef T Mansour, Beatrice Rohrbach, Bruno Vogt, Hiten D Mistry, Geneviève Escher
Sterol 27-hydroxylase (CYP27A1) is involved in bile acid synthesis and cholesterol homoeostasis. Cyp27a1((-/-))/Apolipoprotein E((-/-)) double knockout mice (DKO) fed a western diet failed to develop atherosclerosis. Caveolin-1 (CAV-1), the main component of caveolae, is associated with lipid homoeostasis and has regulatory roles in vascular diseases. We hypothesized that liver CAV-1 would contribute to the athero-protective mechanism in DKO mice. Cyp27a1((+/+))/ApoE((-/-)) (ApoE KO), Cyp27a1((+/-))/ApoE((-/-)) (het), and DKO mice were fed a western diet for 2 months...
October 2016: FEBS Open Bio
https://www.readbyqxmd.com/read/28143572/poly-lactic-acid-nanoparticles-pla-np-promote-physiological-modifications-in-lung-epithelial-cells-and-are-internalized-by-clathrin-coated-pits-and-lipid-rafts
#15
Camila Macedo da Luz, Matthew Samuel Powys Boyles, Priscila Falagan-Lotsch, Mariana Rodrigues Pereira, Henrique Rudolf Tutumi, Eidy de Oliveira Santos, Nathalia Balthazar Martins, Martin Himly, Aniela Sommer, Ilse Foissner, Albert Duschl, José Mauro Granjeiro, Paulo Emílio Corrêa Leite
BACKGROUND: Poly-lactic acid nanoparticles (PLA-NP) are a type of polymeric NP, frequently used as nanomedicines, which have advantages over metallic NP such as the ability to maintain therapeutic drug levels for sustained periods of time. Despite PLA-NP being considered biocompatible, data concerning alterations in cellular physiology are scarce. METHODS: We conducted an extensive evaluation of PLA-NP biocompatibility in human lung epithelial A549 cells using high throughput screening and more complex methodologies...
January 31, 2017: Journal of Nanobiotechnology
https://www.readbyqxmd.com/read/28130355/2intestinal-epithelial-cell-caveolin-1-regulates-fatty-acid-and-lipoprotein-cholesterol-plasma-levels
#16
Jessica P Otis, Meng-Chieh Shen, Vanessa Quinlivan, Jennifer L Anderson, Steven A Farber
Caveolae and their structural protein caveolin 1 (CAV1) have roles in cellular lipid processing and systemic lipid metabolism. Global deletion of CAV1 in mice results in insulin resistance and increases in atherogenic plasma lipids and cholesterol, but protects from diet-induced obesity and atherosclerosis. Despite the fundamental role of the intestinal epithelia in the regulation of dietary lipid processing and metabolism, the contributions of CAV1 to lipid metabolism in this tissue have never been directly investigated...
January 26, 2017: Disease Models & Mechanisms
https://www.readbyqxmd.com/read/28130294/transcellular-pathways-in-lymphatic-endothelial-cells-regulate-changes-in-solute-transport-by-fluid-stress
#17
Valentina Triacca, Esra Guc, Witold W Kilarski, Marco Pisano, Melody A Swartz
RATIONALE: The transport of interstitial fluid and solutes into lymphatic vessels is important for maintaining interstitial homeostasis and delivering antigens and soluble factors to the lymph node for immune surveillance. Transendothelial transport across lymphatic endothelial cells (LECs) is commonly considered to occur paracellularly, or between cell-cell junctions, and driven by local pressure and concentration gradients. However, emerging evidence suggests that LECs also play active roles in regulating interstitial solute balance and can scavenge and store antigens, raising the possibility that vesicular or transcellular pathways may be important in lymphatic solute transport...
January 27, 2017: Circulation Research
https://www.readbyqxmd.com/read/28126510/caveolin1-protein-arginine-methyltransferase1-sirtuin1-axis-as-a-potential-target-against-endothelial-dysfunction
#18
REVIEW
Soniya Charles, Vijay Raj, Jesu Arokiaraj, Kanchana Mala
Endothelial dysfunction (ED), an established response to cardiovascular risk factors, is characterized by increased levels of soluble molecules secreted by endothelial cells (EC). Evidence suggest that ED is an independent predictor of cardiac events and that it is associated with a deficiency in production or bioavailability of nitric oxide (NO) and/or an imbalance in the relative contribution of endothelium-derived relaxing and contracting factors. ED can be reversed by treating cardiovascular risk factors, hence, beyond ambiguity, ED contributes to initiation and progression of atherosclerotic disease...
January 23, 2017: Pharmacological Research: the Official Journal of the Italian Pharmacological Society
https://www.readbyqxmd.com/read/28122734/hugh-davson-distinguished-lectureship-article-caveolins-and-cavins-in-the-trafficking-maturation-and-degradation-of-caveolae-implications-for-cell-physiology
#19
Anna R Busija, Hemal H Patel, Paul A Insel
Caveolins (Cavs) are ~20 kDa scaffolding proteins that assemble as oligomeric complexes in lipid raft domains to form caveolae, flask-shaped plasma membrane (PM) invaginations. Caveolae ("little caves") require lipid-lipid, protein-lipid, and protein-protein interactions that can modulate the localization, conformational stability, ligand affinity, effector specificity, and other functions of proteins that are partners of Cavs. Cavs are assembled into small oligomers in the endoplasmic reticulum (ER), transported to the Golgi for assembly with cholesterol and other oligomers, and then exported to the PM as an intact coat complex...
January 25, 2017: American Journal of Physiology. Cell Physiology
https://www.readbyqxmd.com/read/28116722/assessment-of-caveolae-lipid-rafts-in-isolated-cells
#20
G E Callera, Thiago Bruder-Nascimento, R M Touyz
This chapter outlines protocols to evaluate protein localization, recruitment or phosphorylation levels in cholesterol/sphingolipids-enriched cell membrane domains and recommends experimental designs with pharmacological tolls to evaluate potential cell functions associated with these domains. We emphasize the need for the combination of several approaches towards understanding the protein components and cellular functions attributed to these distinct microdomains.
2017: Methods in Molecular Biology
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