Preservation of human cell bioassay device for the on-site evaluation of environmental waters

2002 ◽  
Vol 46 (11-12) ◽  
pp. 355-362 ◽  
Author(s):  
R. Shoji ◽  
Y. Sakai ◽  
A. Sakoda ◽  
M. Suzuki
Keyword(s):  
River Water ◽  
Low Density Lipoprotein ◽  
Water Quality Management ◽  
Density Lipoprotein ◽  
Hep G2 ◽  
Environmental Waters ◽  
Water Toxicity ◽  
Promising Tool ◽  
Site Evaluation ◽  
Daily Water

We have already developed a novel disposable bioassay device based on the low-density lipoprotein (LDL) uptaking activity of human hepatoblastoma Hep G2 cells in our previous work. However, this device is not readily applicable to evaluate river water toxicity on-site because it cannot be preserved for more than one week. In this work, we developed the method for preservation of the device to enable it to be preserved for at least one month. The device can be supplied to individual environmental sites without any facilities for cell culture. We can evaluate river water toxicity by 2 hours of exposure after thawing. Therefore, this kind of device could be a promising tool for daily water quality management.

Formulating biossay data of chemicals and environmental water

2000 ◽  
Vol 42 (3-4) ◽  
pp. 115-123 ◽  
Author(s):  
R. Shoji ◽  
A. Sakoda ◽  
Y. Sakai ◽  
M. Suzuki
Keyword(s):  
Water Quality ◽  
Low Density Lipoprotein ◽  
Water Environment ◽  
Density Lipoprotein ◽  
Environmental Water ◽  
Toxicity Data ◽  
Human Beings ◽  
Environmental Waters ◽  
Bodies Of Water

The quality of environmental waters such as rivers is often deteriorated by various kinds of trace and unidentified chemicals despite the recent development of sewage systems and wastewater treatment technologies. In addition to contamination by particular toxicants, complex toxicity due to multi-component chemicals could be much more serious. The environmental situation in bodies of water in Japan led us to apply bioassays for monitoring the water quality of environmental waters in order to express the direct and potential toxicity to human beings and ecosystems rather than determinating concentrations of particular chemicals. However, problems arose from the fact that bioassays for pharmaceutical purposes generally required complicated, time-consuming, expert procedures. Also, a methodology for feedback of the resultant toxicity data to water environment management has not been established yet. To this end, we developed a novel bioassay based on the low-density lipoprotein (LDL) uptake activity of human hepatoblastoma cells. The assay enabled us to directly detect the toxicity of environmental waters within 4 hours of exposure. This is a significantly quick and easy procedure as compared to that of conventional bioassays. The toxicity data for 255 selected chemicals and environmental waters obtained by this method were organized by a mathematical equation in order to make those data much more effectively and practically useful to the management of environmental waters. Our methodology represents a promising example of applying bioassays to monitor environmental water quality and generating potential solutions to the toxicity problems encountered.

scholarly journals The effect of (−)-hydroxycitrate on the activity of the low-density-lipoprotein receptor and 3-hydroxy-3-methylglutaryl-CoA reductase levels in the human hepatoma cell line Hep G2

1990 ◽  
Vol 272 (1) ◽  
pp. 181-186 ◽  
Author(s):  
T A Berkhout ◽  
L M Havekes ◽  
N J Pearce ◽  
P H E Groot
Keyword(s):  
Low Density Lipoprotein ◽  
Ldl Receptor ◽  
Density Lipoprotein ◽  
Low Density ◽  
Hep G2 ◽  
Atp Citrate Lyase ◽  
Hmg Coa Reductase ◽  
Synthetic Pathway ◽  
Citrate Lyase ◽  
Coa Reductase

(-)-Hydroxycitrate, a potent inhibitor of ATP citrate-lyase, was tested in Hep G2 cells for effects on cholesterol homoeostasis. After 2.5 h and 18 h incubations with (-)-hydroxycitrate at concentrations of 0.5 mM or higher, incorporation of [1,5-14C]citrate into fatty acids and cholesterol was strongly inhibited. This most likely reflects an effective inhibition of ATP citrate-lyase. Cholesterol biosynthesis was decreased to 27% of the control value as measured by incorporations from 3H2O, indicating a decreased flux of carbon units through the cholesterol-synthetic pathway. After 18 h preincubation with 2 mM-(-)-hydroxycitrate, the cellular low-density-lipoprotein (LDL) receptor activity was increased by 50%, as determined by the receptor-mediated association and degradation. Measurements of receptor-mediated binding versus LDL concentration suggests that this increase was due to an increase in the numbers of LDL receptors. Simultaneously, enzyme levels of 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) reductase as determined by activity measurements increased 30-fold. Our results suggest that the increases in HMG-CoA reductase and the LDL receptor are initiated by the decreased flux of carbon units in the cholesterol-synthetic pathway, owing to inhibition of ATP citratelyase. A similar induction of HMG-CoA reductase and LDL receptor was also found after preincubations of cells with 0.3 microM-mevinolin, suggesting that the underlying mechanism for this induction is identical for both drugs.

Differences in metabolism of three low density lipoprotein subfractions in Hep G2 cells

1990 ◽  
Vol 1047 (3) ◽  
pp. 212-222 ◽  
Author(s):  
Dorine W. Swinkels ◽  
Jan C.M. Hendriks ◽  
Pierre N.M. Demacker ◽  
Anton F.H. Stalenhoef
Keyword(s):  
Low Density Lipoprotein ◽  
Density Lipoprotein ◽  
Low Density ◽  
Hep G2 ◽  
Hep G2 Cells ◽  
Lipoprotein Subfractions ◽  
G2 Cells

Interaction of apolipoprotein B-containing lipoprotein secreted by Hep G2 cells with receptors for low-density lipoprotein

1991 ◽  
Vol 1086 (2) ◽  
pp. 237-240 ◽  
Author(s):  
Ilia V. Fuki ◽  
Georgy B. Menschikov ◽  
Mario Menschikowski ◽  
Irina Yu. Adamova ◽  
Vadim S. Repin
Keyword(s):  
Apolipoprotein B ◽  
Low Density Lipoprotein ◽  
Density Lipoprotein ◽  
Low Density ◽  
Hep G2 ◽  
Hep G2 Cells ◽  
G2 Cells

scholarly journals Characterization of the low-density-lipoprotein-receptor-independent interaction of β-very-low-density lipoprotein with rat and human parenchymal liver cells in vitro

1992 ◽  
Vol 282 (1) ◽  
pp. 41-48 ◽  
Author(s):  
R De Water ◽  
J A A M Kamps ◽  
M C M Van Dijk ◽  
E A M J Hessels ◽  
J Kuiper ◽  
...  
Keyword(s):  
Low Density Lipoprotein ◽  
Ldl Receptor ◽  
Density Lipoprotein ◽  
Liver Cells ◽  
Recognition Site ◽  
Low Density ◽  
Hep G2 ◽  
Hep G2 Cells ◽  
Parenchymal Liver ◽  
Parenchymal Cells

beta-Migrating very-low-density lipoprotein (beta-VLDL) is a cholesteryl-ester-enriched lipoprotein which under normal conditions is rapidly cleared by parenchymal liver cells. In this study the characteristics of the interaction of beta-VLDL with rat parenchymal cells, Hep G2 cells and human parenchymal cells are evaluated. The binding of beta-VLDL to these cells follows saturation kinetics (Bmax. respectively 117, 106 and 103 ng of beta-VLDL apoliprotein/mg of cell protein), with a relatively high affinity (Kd respectively for beta-VLDL of 10.7, 5.1 and 8.4 micrograms/ml). Competition studies of unlabelled beta-VLDL, low-density lipoprotein (LDL) or acetylated LDL with the binding of radiolabelled beta-VLDL indicate that a LDL-receptor-independent, Ca(2+)-independent, specific recognition site for beta-VLDL is present on rat and human parenchymal cells, whereas with Hep G2 cells or mouse macrophages beta-VLDL recognition is performed by the LDL receptor. The binding of beta-VLDL to Hep G2 cells was down-regulated by 89% by prolonged exposure to beta-VLDL, whereas for human parenchymal and rat parenchymal cells down-regulation of 44% and 20% respectively was observed. Studies with antibodies against the LDL receptor support the presence of a LDL-receptor-independent specific beta-VLDL recognition site on rat and human parenchymal cells. It is concluded that a LDL-receptor-independent recognition site for beta-VLDL is present on rat and human parenchymal liver cells. The presence of a LDL-receptor-independent recognition site on human parenchymal cells may mediate in vivo the uptake of beta-VLDL during consumption of a cholesterol-rich diet, when LDL receptors are down-regulated, thus protecting against the extrahepatic accumulation of the atherogenic beta-VLDL constituents.

scholarly journals CRISPR Gene Editing in Lipid Disorders and Atherosclerosis: Mechanisms and Opportunities

Metabolites ◽  
2021 ◽  
Vol 11 (12) ◽  
pp. 857
Author(s):  
Harry E. Walker ◽  
Manfredi Rizzo ◽  
Zlatko Fras ◽  
Borut Jug ◽  
Maciej Banach ◽  
...  
Keyword(s):  
Cardiovascular Disease ◽  
Gene Editing ◽  
Low Density Lipoprotein ◽  
Experimental Studies ◽  
Ldl Receptor ◽  
Density Lipoprotein ◽  
Atherosclerotic Cardiovascular Disease ◽  
Subcutaneous Injections ◽  
Lipid Disorders ◽  
Promising Tool

Elevated circulating concentrations of low-density lipoprotein cholesterol (LDL-C) have been conclusively demonstrated in epidemiological and intervention studies to be causally associated with the development of atherosclerotic cardiovascular disease. Enormous advances in LDL-C reduction have been achieved through the use of statins, and in recent years, through drugs targeting proprotein convertase subtilisin/kexin type 9 (PCSK9), a key regulator of the hepatic LDL-receptor. Existing approaches to PCSK9 targeting have used monoclonal antibodies or RNA interference. Although these approaches do not require daily dosing, as statins do, repeated subcutaneous injections are nevertheless necessary to maintain effectiveness over time. Recent experimental studies suggest that clustered regularly interspaced short palindromic repeats (CRISPR) gene-editing targeted at PCSK9 may represent a promising tool to achieve the elusive goal of a ‘fire and forget’ lifelong approach to LDL-C reduction. This paper will provide an overview of CRISPR technology, with a particular focus on recent studies with relevance to its potential use in atherosclerotic cardiovascular disease.

Comparison of the LDL-receptor binding of VLDL and LDL from apoE4 and apoE3 homozygotes

1999 ◽  
Vol 276 (3) ◽  
pp. E553-E557 ◽  
Author(s):  
Cyril D. S. Mamotte ◽  
Marian Sturm ◽  
Jock I. Foo ◽  
Frank M. van Bockxmeer ◽  
Roger R. Taylor
Keyword(s):  
Plasma Lipids ◽  
Low Density Lipoprotein ◽  
Ldl Receptor ◽  
Density Lipoprotein ◽  
Low Density ◽  
Hep G2 ◽  
Hep G2 Cells ◽  
Cultured Fibroblasts ◽  
Ldl Receptors ◽  
G2 Cells

Compared with apolipoprotein E3 (apoE3), apoE2 is less effective in mediating the binding of lipoproteins to the low-density lipoprotein (LDL) receptor. The influence of the E4 isoform, which is associated with adverse effects on plasma lipids and coronary heart disease, is less clear. We compared the ability of very low density lipoprotein (VLDL) and LDL from paired E4/4 and E3/3 subjects to compete against125I-labeled LDL for binding with the LDL receptor on cultured fibroblasts and Hep G2 cells. The concentrations of VLDL or LDL required to inhibit binding of125I-LDL by 50% (IC50, μg apoB/ml) were determined, and results were assessed in terms of an IC50 ratio, E4/4 IC50 relative to E3/3 IC50, to reduce the influence of interassay variability. In Hep G2 cells, E4/4 VLDL was more effective than E3/3 VLDL in competing for the LDL receptor, the IC50 ratio being lower than unity (0.73 ± 0.31, P < 0.05, two-tailed t-test). IC50 values themselves were marginally lower in E4/4 than E3/3 subjects (3.7 ± 1.3 vs. 6.1 ± 3.7, P < 0.08). However, there was no difference between E4/4 and E3/3 VLDL in competing for the LDL receptor on fibroblasts or between E4/4 and E3/3 LDL in competing for the LDL receptor on either cell type. These results suggest that inheritance of apoE4 is associated with an increased affinity of VLDL particles for LDL receptors on hepatocytes and may partly explain the influence of the E4 isoform on lipid metabolism.

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