Abstract

We report here the observation of the enhancement of Europium-tetracycline complex emission in Low Density Lipoprotein (LDL) solutions. Europium emission band of tetracycline solution containing Europium (III) chloride hexahydrate was tested to obtain effective enhancement in the presence of native LDL and oxidized LDL. Europium emission lifetime in the presence of lipoproteins was measured, resulting in a simple method to measure the lipoproteins quantity in an aqueous solution at physiological pH. This method shows that the complex can be used as a sensor to determine the different states of native and oxidized LDL in biological fluids.

© 2007 Optical Society of America

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  14. N. Arnaud and J. Georges, "Comprehensive study of the luminescent properties and lifetimes of Eu[3][+] and Tb[3][+] chelated with various ligands in aqueous solutions: influence ofthe synergic agent, the surfactant and the energy level of the ligand triplet," Spectrochim. Acta A Mol. Biomol. Spectrosc. 59, 1829-1840 (2003).
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    [CrossRef] [PubMed]
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    [CrossRef] [PubMed]
  23. L.M. Hirschy, T.F. Van Geel, J. D. Winefordner, R.N. Kelly, and S.G. Schulman, "Characteristics of the binding of Europium(III) to Tetracycline," Anal. Chim. Acta. 166, 207-219 (1984).
    [CrossRef]
  24. I. Chopra and M. Roberts, "Tetracycline Antibiotics: Mode of Action, Applications, Molecular Biology, and Epidemiology of Bacterial Resistence," Microb. Mol. Biol. Rev. 65, 232-260 (2001).
    [CrossRef]
  25. T. Ha, T. Enderle, D.S. Chemia, P.R Selvin, S Weiss, "Quantun jumps of single molecules at room temperature," Chem. Phys. Lett. 271, 1-5 (1997).
    [CrossRef]
  26. H. Esterbauer, J. Gebicki, H. Puhl, and G. Jurgens, "The role of lipid peroxidation and antioxidants in oxidative modification of LDL," Free Rad. Biol. Med. 13, 341-390 (1992).
    [CrossRef] [PubMed]
  27. H. Puhl, G. Waeg and H. Esterbauer, "Methods to determine oxidation of low-density lipoproteins," Meth. Enzymol. 233, 425-441 (1994).
  28. M. Wu, Z.H. Lin, A. Durkop, O.S. Wolfbeis, "Time-resolved enzymatic determination of glucose using a fluorescent Europium probe for hydrogen peroxide," Anal. and Bioanal. Chem. 380, 619-626 (2004).
    [CrossRef]
  29. B. Vedie, I. Myara, M.A. Pech, J.C. Maziere, C. Maziere, A. Caprani, and N. Moatti, "Fractionation of chargemodified low density lipoproteins by fast protein liquid chromatography," J. Lipid Research 32, 1359-1369 (1991).

2006 (2)

C. Danese, A.R. Vestri, V. D. Alfonso, G. Deriu, S. Dispensa, R. Baldini, M. Ambrosino, M. Colotto, "Do hypertension and diabetes mellitus influence the site of Atherosclerotic plaques?," Clin. Ter. 157, 9-13 (2006).
[PubMed]

Z. Lin,M. Wu, O.S. Wolfbeis,M. Schaferling, "A novel method for time-resolved fluorimetric determination and imaging of the activity of peroxidase, and its application to an enzyme-linked immunosorbent assay," Chemistry,  12, 2730-2738 (2006).
[CrossRef] [PubMed]

2005 (2)

M. Wu, Z. Lin, M. Schaferling, A. Durkop, O.S. Wolfbeis, "Fluorescence imaging of the activity of glucose oxidase using a hydrogen-peroxide-sensitive Europium probe," Anal Biochem. 340, 66-73 (2005).
[CrossRef] [PubMed]

F. R. da Silva, L. C. Courrol, L. V. Gomes Tarelho, L. Gomes, N. Dias VieiraJ’unior, "Enhancement of Europium luminescence in tetracycline-Europium complexes in the presence of urea hydrogen peroxide," J. Fluoresc. 15, 667-671 (2005).
[CrossRef] [PubMed]

2004 (3)

S.L. G’omez, R.F . Turchiello, M.C. Jurado, P. Boschcov, M. Gidlund and A.M. Figueiredo Neto, Chemistry and Physics of Lipids, "Characterization of Native and Oxidized Human Low-Density Lipoproteins by the Z-scan Technique,"  132, 185-195 (2004).

E.C. Fernvik, D.F.J. Ketelhuth, M. Russo, and M. Gidlund, "The autoantibody repertoire against copper- or macrophage-modified LDL differs in normolipidemics and hypercholesterolemic patients," J. Clin. Immunology 24, 170-176 (2004).
[CrossRef]

M. Wu, Z.H. Lin, A. Durkop, O.S. Wolfbeis, "Time-resolved enzymatic determination of glucose using a fluorescent Europium probe for hydrogen peroxide," Anal. and Bioanal. Chem. 380, 619-626 (2004).
[CrossRef]

2003 (1)

N. Arnaud and J. Georges, "Comprehensive study of the luminescent properties and lifetimes of Eu[3][+] and Tb[3][+] chelated with various ligands in aqueous solutions: influence ofthe synergic agent, the surfactant and the energy level of the ligand triplet," Spectrochim. Acta A Mol. Biomol. Spectrosc. 59, 1829-1840 (2003).
[CrossRef] [PubMed]

2002 (1)

Detection and Evaluation, Circulation, American Heart Association 106, 3227-3234 (2002).

2001 (2)

S. Xu and B. Lin, "The Mechanism of Oxidation-Induced Low-Density Lipoprotein Aggregation: An Analogy to Colloidal Aggregation and Beyond?," Biophys. J. 81, 2403-2413 (2001).
[CrossRef] [PubMed]

I. Chopra and M. Roberts, "Tetracycline Antibiotics: Mode of Action, Applications, Molecular Biology, and Epidemiology of Bacterial Resistence," Microb. Mol. Biol. Rev. 65, 232-260 (2001).
[CrossRef]

2000 (1)

T. Hevonoja, M.O. Pentikäinen, M.T. Hyvönen, P.T. Kovanen and M. Ala-Korpela, "Structure of low density lipoprotein (LDL) particles. Basis for understanding molecular changes in modified LDL," Biochim. Biophys. Acta 1488, 189-210 (2000).
[PubMed]

1999 (1)

Y. Rakicioglu, J. H. Perrin, S. G. Schuman, "Increased luminescence of the tetracycline-Europium(III) system following oxidation by hydrogen peroxide," J. Pharm. and Biom. Analysis 20, 397-399 (1999).
[CrossRef]

1998 (2)

D. O’Neal, P. Harrip, G. Dragicevic, D. Rae, and J. D. Best, "A comparison of LDL size determination using gradient gel electrophoresis and light-scattering method," J. Lipid Research 39, 2086-2090 (1998).

W. Castelli, "Lipoproteins and Cardiovascular Disease: Biological Basis and Epidemiological Studies," Value Health. 1, 105-109, (1998).
[CrossRef]

1997 (1)

T. Ha, T. Enderle, D.S. Chemia, P.R Selvin, S Weiss, "Quantun jumps of single molecules at room temperature," Chem. Phys. Lett. 271, 1-5 (1997).
[CrossRef]

1995 (1)

S. Stemme, B. Faber, J. Holm, O. Wiklund, J.L. Witztum, G.K. Hansson, "T lymphocytes from human atherosclerotic plaques recognize oxidized Low density lipoprotein," Proc. Nat. Acad. Sci. USA 92, 3893-3897 (1995).
[CrossRef] [PubMed]

1994 (2)

P. R. Selvin, J.E. Hearst, "Luminescence Energy Transfer Using a Terbium Chelate: Improvements on Fluorescence Energy Transfer," Proc. Nat. Acad. Sci. USA 91, 10024-10028 (1994).
[CrossRef] [PubMed]

H. Puhl, G. Waeg and H. Esterbauer, "Methods to determine oxidation of low-density lipoproteins," Meth. Enzymol. 233, 425-441 (1994).

1992 (2)

H. Esterbauer, J. Gebicki, H. Puhl, and G. Jurgens, "The role of lipid peroxidation and antioxidants in oxidative modification of LDL," Free Rad. Biol. Med. 13, 341-390 (1992).
[CrossRef] [PubMed]

D.G. Hassall, "Three probe flow cytometry of a human foam-cell forming macrophage," Cytometry 13, 381-388 (1992).
[CrossRef] [PubMed]

1991 (1)

B. Vedie, I. Myara, M.A. Pech, J.C. Maziere, C. Maziere, A. Caprani, and N. Moatti, "Fractionation of chargemodified low density lipoproteins by fast protein liquid chromatography," J. Lipid Research 32, 1359-1369 (1991).

1987 (1)

H. Hara, H. Tanishita, S. Yokoyama, S. Tajima, and A. Yamamoto, "Induction of acetylated low density lipoprotein receptors and suppression of low density liprotein receptors on the cells of human monocytic leukemia cell line (THP-I cell)," Biochem. Biophys. Res. Commun. 146, 802-808 (1987).
[CrossRef] [PubMed]

1985 (1)

J. R. Crouse, J. S. Parks, H. M. Schey, F. R. Kahl, "Studies of low density lipoprotein molecular weight in human beings with coronary artery disease," J. Lipid Research. 26, 566-574 (1985).

1984 (2)

J. Magnam, D. Barthes, J.J. Giraud., "Complexes of tetracyclines. Study of 1:1 complexes of iron(III), cobalt(II), copper(II) and nickel(II) with tetracycline, oxytetracycline, demeclocycline, doxycycline and minocycline," Ann. Pharm. Fr. 42, 155-159 (1984).
[PubMed]

L.M. Hirschy, T.F. Van Geel, J. D. Winefordner, R.N. Kelly, and S.G. Schulman, "Characteristics of the binding of Europium(III) to Tetracycline," Anal. Chim. Acta. 166, 207-219 (1984).
[CrossRef]

Ala-Korpela, M.

T. Hevonoja, M.O. Pentikäinen, M.T. Hyvönen, P.T. Kovanen and M. Ala-Korpela, "Structure of low density lipoprotein (LDL) particles. Basis for understanding molecular changes in modified LDL," Biochim. Biophys. Acta 1488, 189-210 (2000).
[PubMed]

Alfonso, V. D.

C. Danese, A.R. Vestri, V. D. Alfonso, G. Deriu, S. Dispensa, R. Baldini, M. Ambrosino, M. Colotto, "Do hypertension and diabetes mellitus influence the site of Atherosclerotic plaques?," Clin. Ter. 157, 9-13 (2006).
[PubMed]

Ambrosino, M.

C. Danese, A.R. Vestri, V. D. Alfonso, G. Deriu, S. Dispensa, R. Baldini, M. Ambrosino, M. Colotto, "Do hypertension and diabetes mellitus influence the site of Atherosclerotic plaques?," Clin. Ter. 157, 9-13 (2006).
[PubMed]

Arnaud, N.

N. Arnaud and J. Georges, "Comprehensive study of the luminescent properties and lifetimes of Eu[3][+] and Tb[3][+] chelated with various ligands in aqueous solutions: influence ofthe synergic agent, the surfactant and the energy level of the ligand triplet," Spectrochim. Acta A Mol. Biomol. Spectrosc. 59, 1829-1840 (2003).
[CrossRef] [PubMed]

Baldini, R.

C. Danese, A.R. Vestri, V. D. Alfonso, G. Deriu, S. Dispensa, R. Baldini, M. Ambrosino, M. Colotto, "Do hypertension and diabetes mellitus influence the site of Atherosclerotic plaques?," Clin. Ter. 157, 9-13 (2006).
[PubMed]

Barthes, D.

J. Magnam, D. Barthes, J.J. Giraud., "Complexes of tetracyclines. Study of 1:1 complexes of iron(III), cobalt(II), copper(II) and nickel(II) with tetracycline, oxytetracycline, demeclocycline, doxycycline and minocycline," Ann. Pharm. Fr. 42, 155-159 (1984).
[PubMed]

Best, J. D.

D. O’Neal, P. Harrip, G. Dragicevic, D. Rae, and J. D. Best, "A comparison of LDL size determination using gradient gel electrophoresis and light-scattering method," J. Lipid Research 39, 2086-2090 (1998).

Boschcov, P.

S.L. G’omez, R.F . Turchiello, M.C. Jurado, P. Boschcov, M. Gidlund and A.M. Figueiredo Neto, Chemistry and Physics of Lipids, "Characterization of Native and Oxidized Human Low-Density Lipoproteins by the Z-scan Technique,"  132, 185-195 (2004).

Caprani, A.

B. Vedie, I. Myara, M.A. Pech, J.C. Maziere, C. Maziere, A. Caprani, and N. Moatti, "Fractionation of chargemodified low density lipoproteins by fast protein liquid chromatography," J. Lipid Research 32, 1359-1369 (1991).

Castelli, W.

W. Castelli, "Lipoproteins and Cardiovascular Disease: Biological Basis and Epidemiological Studies," Value Health. 1, 105-109, (1998).
[CrossRef]

Chemia, D.S.

T. Ha, T. Enderle, D.S. Chemia, P.R Selvin, S Weiss, "Quantun jumps of single molecules at room temperature," Chem. Phys. Lett. 271, 1-5 (1997).
[CrossRef]

Chopra, I.

I. Chopra and M. Roberts, "Tetracycline Antibiotics: Mode of Action, Applications, Molecular Biology, and Epidemiology of Bacterial Resistence," Microb. Mol. Biol. Rev. 65, 232-260 (2001).
[CrossRef]

Colotto, M.

C. Danese, A.R. Vestri, V. D. Alfonso, G. Deriu, S. Dispensa, R. Baldini, M. Ambrosino, M. Colotto, "Do hypertension and diabetes mellitus influence the site of Atherosclerotic plaques?," Clin. Ter. 157, 9-13 (2006).
[PubMed]

Courrol, L. C.

F. R. da Silva, L. C. Courrol, L. V. Gomes Tarelho, L. Gomes, N. Dias VieiraJ’unior, "Enhancement of Europium luminescence in tetracycline-Europium complexes in the presence of urea hydrogen peroxide," J. Fluoresc. 15, 667-671 (2005).
[CrossRef] [PubMed]

Crouse, J. R.

J. R. Crouse, J. S. Parks, H. M. Schey, F. R. Kahl, "Studies of low density lipoprotein molecular weight in human beings with coronary artery disease," J. Lipid Research. 26, 566-574 (1985).

da Silva, F. R.

F. R. da Silva, L. C. Courrol, L. V. Gomes Tarelho, L. Gomes, N. Dias VieiraJ’unior, "Enhancement of Europium luminescence in tetracycline-Europium complexes in the presence of urea hydrogen peroxide," J. Fluoresc. 15, 667-671 (2005).
[CrossRef] [PubMed]

Danese, C.

C. Danese, A.R. Vestri, V. D. Alfonso, G. Deriu, S. Dispensa, R. Baldini, M. Ambrosino, M. Colotto, "Do hypertension and diabetes mellitus influence the site of Atherosclerotic plaques?," Clin. Ter. 157, 9-13 (2006).
[PubMed]

Deriu, G.

C. Danese, A.R. Vestri, V. D. Alfonso, G. Deriu, S. Dispensa, R. Baldini, M. Ambrosino, M. Colotto, "Do hypertension and diabetes mellitus influence the site of Atherosclerotic plaques?," Clin. Ter. 157, 9-13 (2006).
[PubMed]

Dias Vieira, N.

F. R. da Silva, L. C. Courrol, L. V. Gomes Tarelho, L. Gomes, N. Dias VieiraJ’unior, "Enhancement of Europium luminescence in tetracycline-Europium complexes in the presence of urea hydrogen peroxide," J. Fluoresc. 15, 667-671 (2005).
[CrossRef] [PubMed]

Dispensa, S.

C. Danese, A.R. Vestri, V. D. Alfonso, G. Deriu, S. Dispensa, R. Baldini, M. Ambrosino, M. Colotto, "Do hypertension and diabetes mellitus influence the site of Atherosclerotic plaques?," Clin. Ter. 157, 9-13 (2006).
[PubMed]

Dragicevic, G.

D. O’Neal, P. Harrip, G. Dragicevic, D. Rae, and J. D. Best, "A comparison of LDL size determination using gradient gel electrophoresis and light-scattering method," J. Lipid Research 39, 2086-2090 (1998).

Durkop, A.

M. Wu, Z. Lin, M. Schaferling, A. Durkop, O.S. Wolfbeis, "Fluorescence imaging of the activity of glucose oxidase using a hydrogen-peroxide-sensitive Europium probe," Anal Biochem. 340, 66-73 (2005).
[CrossRef] [PubMed]

M. Wu, Z.H. Lin, A. Durkop, O.S. Wolfbeis, "Time-resolved enzymatic determination of glucose using a fluorescent Europium probe for hydrogen peroxide," Anal. and Bioanal. Chem. 380, 619-626 (2004).
[CrossRef]

Enderle, T.

T. Ha, T. Enderle, D.S. Chemia, P.R Selvin, S Weiss, "Quantun jumps of single molecules at room temperature," Chem. Phys. Lett. 271, 1-5 (1997).
[CrossRef]

Esterbauer, H.

H. Puhl, G. Waeg and H. Esterbauer, "Methods to determine oxidation of low-density lipoproteins," Meth. Enzymol. 233, 425-441 (1994).

H. Esterbauer, J. Gebicki, H. Puhl, and G. Jurgens, "The role of lipid peroxidation and antioxidants in oxidative modification of LDL," Free Rad. Biol. Med. 13, 341-390 (1992).
[CrossRef] [PubMed]

Faber, B.

S. Stemme, B. Faber, J. Holm, O. Wiklund, J.L. Witztum, G.K. Hansson, "T lymphocytes from human atherosclerotic plaques recognize oxidized Low density lipoprotein," Proc. Nat. Acad. Sci. USA 92, 3893-3897 (1995).
[CrossRef] [PubMed]

Fernvik, E.C.

E.C. Fernvik, D.F.J. Ketelhuth, M. Russo, and M. Gidlund, "The autoantibody repertoire against copper- or macrophage-modified LDL differs in normolipidemics and hypercholesterolemic patients," J. Clin. Immunology 24, 170-176 (2004).
[CrossRef]

Figueiredo Neto, A.M.

S.L. G’omez, R.F . Turchiello, M.C. Jurado, P. Boschcov, M. Gidlund and A.M. Figueiredo Neto, Chemistry and Physics of Lipids, "Characterization of Native and Oxidized Human Low-Density Lipoproteins by the Z-scan Technique,"  132, 185-195 (2004).

G’omez, S.L.

S.L. G’omez, R.F . Turchiello, M.C. Jurado, P. Boschcov, M. Gidlund and A.M. Figueiredo Neto, Chemistry and Physics of Lipids, "Characterization of Native and Oxidized Human Low-Density Lipoproteins by the Z-scan Technique,"  132, 185-195 (2004).

Gebicki, J.

H. Esterbauer, J. Gebicki, H. Puhl, and G. Jurgens, "The role of lipid peroxidation and antioxidants in oxidative modification of LDL," Free Rad. Biol. Med. 13, 341-390 (1992).
[CrossRef] [PubMed]

Georges, J.

N. Arnaud and J. Georges, "Comprehensive study of the luminescent properties and lifetimes of Eu[3][+] and Tb[3][+] chelated with various ligands in aqueous solutions: influence ofthe synergic agent, the surfactant and the energy level of the ligand triplet," Spectrochim. Acta A Mol. Biomol. Spectrosc. 59, 1829-1840 (2003).
[CrossRef] [PubMed]

Gidlund, M.

S.L. G’omez, R.F . Turchiello, M.C. Jurado, P. Boschcov, M. Gidlund and A.M. Figueiredo Neto, Chemistry and Physics of Lipids, "Characterization of Native and Oxidized Human Low-Density Lipoproteins by the Z-scan Technique,"  132, 185-195 (2004).

E.C. Fernvik, D.F.J. Ketelhuth, M. Russo, and M. Gidlund, "The autoantibody repertoire against copper- or macrophage-modified LDL differs in normolipidemics and hypercholesterolemic patients," J. Clin. Immunology 24, 170-176 (2004).
[CrossRef]

Giraud, J.J.

J. Magnam, D. Barthes, J.J. Giraud., "Complexes of tetracyclines. Study of 1:1 complexes of iron(III), cobalt(II), copper(II) and nickel(II) with tetracycline, oxytetracycline, demeclocycline, doxycycline and minocycline," Ann. Pharm. Fr. 42, 155-159 (1984).
[PubMed]

Gomes, L.

F. R. da Silva, L. C. Courrol, L. V. Gomes Tarelho, L. Gomes, N. Dias VieiraJ’unior, "Enhancement of Europium luminescence in tetracycline-Europium complexes in the presence of urea hydrogen peroxide," J. Fluoresc. 15, 667-671 (2005).
[CrossRef] [PubMed]

Gomes Tarelho, L. V.

F. R. da Silva, L. C. Courrol, L. V. Gomes Tarelho, L. Gomes, N. Dias VieiraJ’unior, "Enhancement of Europium luminescence in tetracycline-Europium complexes in the presence of urea hydrogen peroxide," J. Fluoresc. 15, 667-671 (2005).
[CrossRef] [PubMed]

Ha, T.

T. Ha, T. Enderle, D.S. Chemia, P.R Selvin, S Weiss, "Quantun jumps of single molecules at room temperature," Chem. Phys. Lett. 271, 1-5 (1997).
[CrossRef]

Hansson, G.K.

S. Stemme, B. Faber, J. Holm, O. Wiklund, J.L. Witztum, G.K. Hansson, "T lymphocytes from human atherosclerotic plaques recognize oxidized Low density lipoprotein," Proc. Nat. Acad. Sci. USA 92, 3893-3897 (1995).
[CrossRef] [PubMed]

Hara, H.

H. Hara, H. Tanishita, S. Yokoyama, S. Tajima, and A. Yamamoto, "Induction of acetylated low density lipoprotein receptors and suppression of low density liprotein receptors on the cells of human monocytic leukemia cell line (THP-I cell)," Biochem. Biophys. Res. Commun. 146, 802-808 (1987).
[CrossRef] [PubMed]

Harrip, P.

D. O’Neal, P. Harrip, G. Dragicevic, D. Rae, and J. D. Best, "A comparison of LDL size determination using gradient gel electrophoresis and light-scattering method," J. Lipid Research 39, 2086-2090 (1998).

Hassall, D.G.

D.G. Hassall, "Three probe flow cytometry of a human foam-cell forming macrophage," Cytometry 13, 381-388 (1992).
[CrossRef] [PubMed]

Hearst, J.E.

P. R. Selvin, J.E. Hearst, "Luminescence Energy Transfer Using a Terbium Chelate: Improvements on Fluorescence Energy Transfer," Proc. Nat. Acad. Sci. USA 91, 10024-10028 (1994).
[CrossRef] [PubMed]

Hevonoja, T.

T. Hevonoja, M.O. Pentikäinen, M.T. Hyvönen, P.T. Kovanen and M. Ala-Korpela, "Structure of low density lipoprotein (LDL) particles. Basis for understanding molecular changes in modified LDL," Biochim. Biophys. Acta 1488, 189-210 (2000).
[PubMed]

Hirschy, L.M.

L.M. Hirschy, T.F. Van Geel, J. D. Winefordner, R.N. Kelly, and S.G. Schulman, "Characteristics of the binding of Europium(III) to Tetracycline," Anal. Chim. Acta. 166, 207-219 (1984).
[CrossRef]

Holm, J.

S. Stemme, B. Faber, J. Holm, O. Wiklund, J.L. Witztum, G.K. Hansson, "T lymphocytes from human atherosclerotic plaques recognize oxidized Low density lipoprotein," Proc. Nat. Acad. Sci. USA 92, 3893-3897 (1995).
[CrossRef] [PubMed]

Hyvönen, M.T.

T. Hevonoja, M.O. Pentikäinen, M.T. Hyvönen, P.T. Kovanen and M. Ala-Korpela, "Structure of low density lipoprotein (LDL) particles. Basis for understanding molecular changes in modified LDL," Biochim. Biophys. Acta 1488, 189-210 (2000).
[PubMed]

Jurado, M.C.

S.L. G’omez, R.F . Turchiello, M.C. Jurado, P. Boschcov, M. Gidlund and A.M. Figueiredo Neto, Chemistry and Physics of Lipids, "Characterization of Native and Oxidized Human Low-Density Lipoproteins by the Z-scan Technique,"  132, 185-195 (2004).

Jurgens, G.

H. Esterbauer, J. Gebicki, H. Puhl, and G. Jurgens, "The role of lipid peroxidation and antioxidants in oxidative modification of LDL," Free Rad. Biol. Med. 13, 341-390 (1992).
[CrossRef] [PubMed]

Kahl, F. R.

J. R. Crouse, J. S. Parks, H. M. Schey, F. R. Kahl, "Studies of low density lipoprotein molecular weight in human beings with coronary artery disease," J. Lipid Research. 26, 566-574 (1985).

Kelly, R.N.

L.M. Hirschy, T.F. Van Geel, J. D. Winefordner, R.N. Kelly, and S.G. Schulman, "Characteristics of the binding of Europium(III) to Tetracycline," Anal. Chim. Acta. 166, 207-219 (1984).
[CrossRef]

Ketelhuth, D.F.J.

E.C. Fernvik, D.F.J. Ketelhuth, M. Russo, and M. Gidlund, "The autoantibody repertoire against copper- or macrophage-modified LDL differs in normolipidemics and hypercholesterolemic patients," J. Clin. Immunology 24, 170-176 (2004).
[CrossRef]

Kovanen, P.T.

T. Hevonoja, M.O. Pentikäinen, M.T. Hyvönen, P.T. Kovanen and M. Ala-Korpela, "Structure of low density lipoprotein (LDL) particles. Basis for understanding molecular changes in modified LDL," Biochim. Biophys. Acta 1488, 189-210 (2000).
[PubMed]

Lin, B.

S. Xu and B. Lin, "The Mechanism of Oxidation-Induced Low-Density Lipoprotein Aggregation: An Analogy to Colloidal Aggregation and Beyond?," Biophys. J. 81, 2403-2413 (2001).
[CrossRef] [PubMed]

Lin, Z.

Z. Lin,M. Wu, O.S. Wolfbeis,M. Schaferling, "A novel method for time-resolved fluorimetric determination and imaging of the activity of peroxidase, and its application to an enzyme-linked immunosorbent assay," Chemistry,  12, 2730-2738 (2006).
[CrossRef] [PubMed]

M. Wu, Z. Lin, M. Schaferling, A. Durkop, O.S. Wolfbeis, "Fluorescence imaging of the activity of glucose oxidase using a hydrogen-peroxide-sensitive Europium probe," Anal Biochem. 340, 66-73 (2005).
[CrossRef] [PubMed]

Lin, Z.H.

M. Wu, Z.H. Lin, A. Durkop, O.S. Wolfbeis, "Time-resolved enzymatic determination of glucose using a fluorescent Europium probe for hydrogen peroxide," Anal. and Bioanal. Chem. 380, 619-626 (2004).
[CrossRef]

Magnam, J.

J. Magnam, D. Barthes, J.J. Giraud., "Complexes of tetracyclines. Study of 1:1 complexes of iron(III), cobalt(II), copper(II) and nickel(II) with tetracycline, oxytetracycline, demeclocycline, doxycycline and minocycline," Ann. Pharm. Fr. 42, 155-159 (1984).
[PubMed]

Maziere, C.

B. Vedie, I. Myara, M.A. Pech, J.C. Maziere, C. Maziere, A. Caprani, and N. Moatti, "Fractionation of chargemodified low density lipoproteins by fast protein liquid chromatography," J. Lipid Research 32, 1359-1369 (1991).

Maziere, J.C.

B. Vedie, I. Myara, M.A. Pech, J.C. Maziere, C. Maziere, A. Caprani, and N. Moatti, "Fractionation of chargemodified low density lipoproteins by fast protein liquid chromatography," J. Lipid Research 32, 1359-1369 (1991).

Moatti, N.

B. Vedie, I. Myara, M.A. Pech, J.C. Maziere, C. Maziere, A. Caprani, and N. Moatti, "Fractionation of chargemodified low density lipoproteins by fast protein liquid chromatography," J. Lipid Research 32, 1359-1369 (1991).

Myara, I.

B. Vedie, I. Myara, M.A. Pech, J.C. Maziere, C. Maziere, A. Caprani, and N. Moatti, "Fractionation of chargemodified low density lipoproteins by fast protein liquid chromatography," J. Lipid Research 32, 1359-1369 (1991).

O’Neal, D.

D. O’Neal, P. Harrip, G. Dragicevic, D. Rae, and J. D. Best, "A comparison of LDL size determination using gradient gel electrophoresis and light-scattering method," J. Lipid Research 39, 2086-2090 (1998).

Parks, J. S.

J. R. Crouse, J. S. Parks, H. M. Schey, F. R. Kahl, "Studies of low density lipoprotein molecular weight in human beings with coronary artery disease," J. Lipid Research. 26, 566-574 (1985).

Pech, M.A.

B. Vedie, I. Myara, M.A. Pech, J.C. Maziere, C. Maziere, A. Caprani, and N. Moatti, "Fractionation of chargemodified low density lipoproteins by fast protein liquid chromatography," J. Lipid Research 32, 1359-1369 (1991).

Pentikäinen, M.O.

T. Hevonoja, M.O. Pentikäinen, M.T. Hyvönen, P.T. Kovanen and M. Ala-Korpela, "Structure of low density lipoprotein (LDL) particles. Basis for understanding molecular changes in modified LDL," Biochim. Biophys. Acta 1488, 189-210 (2000).
[PubMed]

Perrin, J. H.

Y. Rakicioglu, J. H. Perrin, S. G. Schuman, "Increased luminescence of the tetracycline-Europium(III) system following oxidation by hydrogen peroxide," J. Pharm. and Biom. Analysis 20, 397-399 (1999).
[CrossRef]

Puhl, H.

H. Puhl, G. Waeg and H. Esterbauer, "Methods to determine oxidation of low-density lipoproteins," Meth. Enzymol. 233, 425-441 (1994).

H. Esterbauer, J. Gebicki, H. Puhl, and G. Jurgens, "The role of lipid peroxidation and antioxidants in oxidative modification of LDL," Free Rad. Biol. Med. 13, 341-390 (1992).
[CrossRef] [PubMed]

Rae, D.

D. O’Neal, P. Harrip, G. Dragicevic, D. Rae, and J. D. Best, "A comparison of LDL size determination using gradient gel electrophoresis and light-scattering method," J. Lipid Research 39, 2086-2090 (1998).

Rakicioglu, Y.

Y. Rakicioglu, J. H. Perrin, S. G. Schuman, "Increased luminescence of the tetracycline-Europium(III) system following oxidation by hydrogen peroxide," J. Pharm. and Biom. Analysis 20, 397-399 (1999).
[CrossRef]

Roberts, M.

I. Chopra and M. Roberts, "Tetracycline Antibiotics: Mode of Action, Applications, Molecular Biology, and Epidemiology of Bacterial Resistence," Microb. Mol. Biol. Rev. 65, 232-260 (2001).
[CrossRef]

Russo, M.

E.C. Fernvik, D.F.J. Ketelhuth, M. Russo, and M. Gidlund, "The autoantibody repertoire against copper- or macrophage-modified LDL differs in normolipidemics and hypercholesterolemic patients," J. Clin. Immunology 24, 170-176 (2004).
[CrossRef]

Schaferling, M.

Z. Lin,M. Wu, O.S. Wolfbeis,M. Schaferling, "A novel method for time-resolved fluorimetric determination and imaging of the activity of peroxidase, and its application to an enzyme-linked immunosorbent assay," Chemistry,  12, 2730-2738 (2006).
[CrossRef] [PubMed]

M. Wu, Z. Lin, M. Schaferling, A. Durkop, O.S. Wolfbeis, "Fluorescence imaging of the activity of glucose oxidase using a hydrogen-peroxide-sensitive Europium probe," Anal Biochem. 340, 66-73 (2005).
[CrossRef] [PubMed]

Schey, H. M.

J. R. Crouse, J. S. Parks, H. M. Schey, F. R. Kahl, "Studies of low density lipoprotein molecular weight in human beings with coronary artery disease," J. Lipid Research. 26, 566-574 (1985).

Schulman, S.G.

L.M. Hirschy, T.F. Van Geel, J. D. Winefordner, R.N. Kelly, and S.G. Schulman, "Characteristics of the binding of Europium(III) to Tetracycline," Anal. Chim. Acta. 166, 207-219 (1984).
[CrossRef]

Schuman, S. G.

Y. Rakicioglu, J. H. Perrin, S. G. Schuman, "Increased luminescence of the tetracycline-Europium(III) system following oxidation by hydrogen peroxide," J. Pharm. and Biom. Analysis 20, 397-399 (1999).
[CrossRef]

Selvin, P. R.

P. R. Selvin, J.E. Hearst, "Luminescence Energy Transfer Using a Terbium Chelate: Improvements on Fluorescence Energy Transfer," Proc. Nat. Acad. Sci. USA 91, 10024-10028 (1994).
[CrossRef] [PubMed]

Selvin, P.R

T. Ha, T. Enderle, D.S. Chemia, P.R Selvin, S Weiss, "Quantun jumps of single molecules at room temperature," Chem. Phys. Lett. 271, 1-5 (1997).
[CrossRef]

Stemme, S.

S. Stemme, B. Faber, J. Holm, O. Wiklund, J.L. Witztum, G.K. Hansson, "T lymphocytes from human atherosclerotic plaques recognize oxidized Low density lipoprotein," Proc. Nat. Acad. Sci. USA 92, 3893-3897 (1995).
[CrossRef] [PubMed]

Tajima, S.

H. Hara, H. Tanishita, S. Yokoyama, S. Tajima, and A. Yamamoto, "Induction of acetylated low density lipoprotein receptors and suppression of low density liprotein receptors on the cells of human monocytic leukemia cell line (THP-I cell)," Biochem. Biophys. Res. Commun. 146, 802-808 (1987).
[CrossRef] [PubMed]

Tanishita, H.

H. Hara, H. Tanishita, S. Yokoyama, S. Tajima, and A. Yamamoto, "Induction of acetylated low density lipoprotein receptors and suppression of low density liprotein receptors on the cells of human monocytic leukemia cell line (THP-I cell)," Biochem. Biophys. Res. Commun. 146, 802-808 (1987).
[CrossRef] [PubMed]

Turchiello, R.F

S.L. G’omez, R.F . Turchiello, M.C. Jurado, P. Boschcov, M. Gidlund and A.M. Figueiredo Neto, Chemistry and Physics of Lipids, "Characterization of Native and Oxidized Human Low-Density Lipoproteins by the Z-scan Technique,"  132, 185-195 (2004).

Van Geel, T.F.

L.M. Hirschy, T.F. Van Geel, J. D. Winefordner, R.N. Kelly, and S.G. Schulman, "Characteristics of the binding of Europium(III) to Tetracycline," Anal. Chim. Acta. 166, 207-219 (1984).
[CrossRef]

Vedie, B.

B. Vedie, I. Myara, M.A. Pech, J.C. Maziere, C. Maziere, A. Caprani, and N. Moatti, "Fractionation of chargemodified low density lipoproteins by fast protein liquid chromatography," J. Lipid Research 32, 1359-1369 (1991).

Vestri, A.R.

C. Danese, A.R. Vestri, V. D. Alfonso, G. Deriu, S. Dispensa, R. Baldini, M. Ambrosino, M. Colotto, "Do hypertension and diabetes mellitus influence the site of Atherosclerotic plaques?," Clin. Ter. 157, 9-13 (2006).
[PubMed]

Waeg, G.

H. Puhl, G. Waeg and H. Esterbauer, "Methods to determine oxidation of low-density lipoproteins," Meth. Enzymol. 233, 425-441 (1994).

Weiss, S

T. Ha, T. Enderle, D.S. Chemia, P.R Selvin, S Weiss, "Quantun jumps of single molecules at room temperature," Chem. Phys. Lett. 271, 1-5 (1997).
[CrossRef]

Wiklund, O.

S. Stemme, B. Faber, J. Holm, O. Wiklund, J.L. Witztum, G.K. Hansson, "T lymphocytes from human atherosclerotic plaques recognize oxidized Low density lipoprotein," Proc. Nat. Acad. Sci. USA 92, 3893-3897 (1995).
[CrossRef] [PubMed]

Winefordner, J. D.

L.M. Hirschy, T.F. Van Geel, J. D. Winefordner, R.N. Kelly, and S.G. Schulman, "Characteristics of the binding of Europium(III) to Tetracycline," Anal. Chim. Acta. 166, 207-219 (1984).
[CrossRef]

Witztum, J.L.

S. Stemme, B. Faber, J. Holm, O. Wiklund, J.L. Witztum, G.K. Hansson, "T lymphocytes from human atherosclerotic plaques recognize oxidized Low density lipoprotein," Proc. Nat. Acad. Sci. USA 92, 3893-3897 (1995).
[CrossRef] [PubMed]

Wolfbeis, O.S.

Z. Lin,M. Wu, O.S. Wolfbeis,M. Schaferling, "A novel method for time-resolved fluorimetric determination and imaging of the activity of peroxidase, and its application to an enzyme-linked immunosorbent assay," Chemistry,  12, 2730-2738 (2006).
[CrossRef] [PubMed]

M. Wu, Z. Lin, M. Schaferling, A. Durkop, O.S. Wolfbeis, "Fluorescence imaging of the activity of glucose oxidase using a hydrogen-peroxide-sensitive Europium probe," Anal Biochem. 340, 66-73 (2005).
[CrossRef] [PubMed]

M. Wu, Z.H. Lin, A. Durkop, O.S. Wolfbeis, "Time-resolved enzymatic determination of glucose using a fluorescent Europium probe for hydrogen peroxide," Anal. and Bioanal. Chem. 380, 619-626 (2004).
[CrossRef]

Wu, M.

Z. Lin,M. Wu, O.S. Wolfbeis,M. Schaferling, "A novel method for time-resolved fluorimetric determination and imaging of the activity of peroxidase, and its application to an enzyme-linked immunosorbent assay," Chemistry,  12, 2730-2738 (2006).
[CrossRef] [PubMed]

M. Wu, Z. Lin, M. Schaferling, A. Durkop, O.S. Wolfbeis, "Fluorescence imaging of the activity of glucose oxidase using a hydrogen-peroxide-sensitive Europium probe," Anal Biochem. 340, 66-73 (2005).
[CrossRef] [PubMed]

M. Wu, Z.H. Lin, A. Durkop, O.S. Wolfbeis, "Time-resolved enzymatic determination of glucose using a fluorescent Europium probe for hydrogen peroxide," Anal. and Bioanal. Chem. 380, 619-626 (2004).
[CrossRef]

Xu, S.

S. Xu and B. Lin, "The Mechanism of Oxidation-Induced Low-Density Lipoprotein Aggregation: An Analogy to Colloidal Aggregation and Beyond?," Biophys. J. 81, 2403-2413 (2001).
[CrossRef] [PubMed]

Yamamoto, A.

H. Hara, H. Tanishita, S. Yokoyama, S. Tajima, and A. Yamamoto, "Induction of acetylated low density lipoprotein receptors and suppression of low density liprotein receptors on the cells of human monocytic leukemia cell line (THP-I cell)," Biochem. Biophys. Res. Commun. 146, 802-808 (1987).
[CrossRef] [PubMed]

Yokoyama, S.

H. Hara, H. Tanishita, S. Yokoyama, S. Tajima, and A. Yamamoto, "Induction of acetylated low density lipoprotein receptors and suppression of low density liprotein receptors on the cells of human monocytic leukemia cell line (THP-I cell)," Biochem. Biophys. Res. Commun. 146, 802-808 (1987).
[CrossRef] [PubMed]

American Heart Association (1)

Detection and Evaluation, Circulation, American Heart Association 106, 3227-3234 (2002).

Anal Biochem. (1)

M. Wu, Z. Lin, M. Schaferling, A. Durkop, O.S. Wolfbeis, "Fluorescence imaging of the activity of glucose oxidase using a hydrogen-peroxide-sensitive Europium probe," Anal Biochem. 340, 66-73 (2005).
[CrossRef] [PubMed]

Anal. and Bioanal. Chem. (1)

M. Wu, Z.H. Lin, A. Durkop, O.S. Wolfbeis, "Time-resolved enzymatic determination of glucose using a fluorescent Europium probe for hydrogen peroxide," Anal. and Bioanal. Chem. 380, 619-626 (2004).
[CrossRef]

Anal. Chim. Acta. (1)

L.M. Hirschy, T.F. Van Geel, J. D. Winefordner, R.N. Kelly, and S.G. Schulman, "Characteristics of the binding of Europium(III) to Tetracycline," Anal. Chim. Acta. 166, 207-219 (1984).
[CrossRef]

Ann. Pharm. Fr. (1)

J. Magnam, D. Barthes, J.J. Giraud., "Complexes of tetracyclines. Study of 1:1 complexes of iron(III), cobalt(II), copper(II) and nickel(II) with tetracycline, oxytetracycline, demeclocycline, doxycycline and minocycline," Ann. Pharm. Fr. 42, 155-159 (1984).
[PubMed]

Biochem. Biophys. Res. Commun. (1)

H. Hara, H. Tanishita, S. Yokoyama, S. Tajima, and A. Yamamoto, "Induction of acetylated low density lipoprotein receptors and suppression of low density liprotein receptors on the cells of human monocytic leukemia cell line (THP-I cell)," Biochem. Biophys. Res. Commun. 146, 802-808 (1987).
[CrossRef] [PubMed]

Biochim. Biophys. Acta (1)

T. Hevonoja, M.O. Pentikäinen, M.T. Hyvönen, P.T. Kovanen and M. Ala-Korpela, "Structure of low density lipoprotein (LDL) particles. Basis for understanding molecular changes in modified LDL," Biochim. Biophys. Acta 1488, 189-210 (2000).
[PubMed]

Biophys. J. (1)

S. Xu and B. Lin, "The Mechanism of Oxidation-Induced Low-Density Lipoprotein Aggregation: An Analogy to Colloidal Aggregation and Beyond?," Biophys. J. 81, 2403-2413 (2001).
[CrossRef] [PubMed]

Characterization of Native and Oxidized Human Low-Density Lipoproteins by the Z-scan Technique (1)

S.L. G’omez, R.F . Turchiello, M.C. Jurado, P. Boschcov, M. Gidlund and A.M. Figueiredo Neto, Chemistry and Physics of Lipids, "Characterization of Native and Oxidized Human Low-Density Lipoproteins by the Z-scan Technique,"  132, 185-195 (2004).

Chem. Phys. Lett. (1)

T. Ha, T. Enderle, D.S. Chemia, P.R Selvin, S Weiss, "Quantun jumps of single molecules at room temperature," Chem. Phys. Lett. 271, 1-5 (1997).
[CrossRef]

Chemistry (1)

Z. Lin,M. Wu, O.S. Wolfbeis,M. Schaferling, "A novel method for time-resolved fluorimetric determination and imaging of the activity of peroxidase, and its application to an enzyme-linked immunosorbent assay," Chemistry,  12, 2730-2738 (2006).
[CrossRef] [PubMed]

Clin. Ter. (1)

C. Danese, A.R. Vestri, V. D. Alfonso, G. Deriu, S. Dispensa, R. Baldini, M. Ambrosino, M. Colotto, "Do hypertension and diabetes mellitus influence the site of Atherosclerotic plaques?," Clin. Ter. 157, 9-13 (2006).
[PubMed]

Cytometry (1)

D.G. Hassall, "Three probe flow cytometry of a human foam-cell forming macrophage," Cytometry 13, 381-388 (1992).
[CrossRef] [PubMed]

Free Rad. Biol. Med. (1)

H. Esterbauer, J. Gebicki, H. Puhl, and G. Jurgens, "The role of lipid peroxidation and antioxidants in oxidative modification of LDL," Free Rad. Biol. Med. 13, 341-390 (1992).
[CrossRef] [PubMed]

J. Clin. Immunology (1)

E.C. Fernvik, D.F.J. Ketelhuth, M. Russo, and M. Gidlund, "The autoantibody repertoire against copper- or macrophage-modified LDL differs in normolipidemics and hypercholesterolemic patients," J. Clin. Immunology 24, 170-176 (2004).
[CrossRef]

J. Fluoresc. (1)

F. R. da Silva, L. C. Courrol, L. V. Gomes Tarelho, L. Gomes, N. Dias VieiraJ’unior, "Enhancement of Europium luminescence in tetracycline-Europium complexes in the presence of urea hydrogen peroxide," J. Fluoresc. 15, 667-671 (2005).
[CrossRef] [PubMed]

J. Lipid Research (2)

D. O’Neal, P. Harrip, G. Dragicevic, D. Rae, and J. D. Best, "A comparison of LDL size determination using gradient gel electrophoresis and light-scattering method," J. Lipid Research 39, 2086-2090 (1998).

B. Vedie, I. Myara, M.A. Pech, J.C. Maziere, C. Maziere, A. Caprani, and N. Moatti, "Fractionation of chargemodified low density lipoproteins by fast protein liquid chromatography," J. Lipid Research 32, 1359-1369 (1991).

J. Lipid Research. (1)

J. R. Crouse, J. S. Parks, H. M. Schey, F. R. Kahl, "Studies of low density lipoprotein molecular weight in human beings with coronary artery disease," J. Lipid Research. 26, 566-574 (1985).

J. Pharm. and Biom. Analysis (1)

Y. Rakicioglu, J. H. Perrin, S. G. Schuman, "Increased luminescence of the tetracycline-Europium(III) system following oxidation by hydrogen peroxide," J. Pharm. and Biom. Analysis 20, 397-399 (1999).
[CrossRef]

Meth. Enzymol. (1)

H. Puhl, G. Waeg and H. Esterbauer, "Methods to determine oxidation of low-density lipoproteins," Meth. Enzymol. 233, 425-441 (1994).

Microb. Mol. Biol. Rev. (1)

I. Chopra and M. Roberts, "Tetracycline Antibiotics: Mode of Action, Applications, Molecular Biology, and Epidemiology of Bacterial Resistence," Microb. Mol. Biol. Rev. 65, 232-260 (2001).
[CrossRef]

Proc. Nat. Acad. Sci. USA (2)

P. R. Selvin, J.E. Hearst, "Luminescence Energy Transfer Using a Terbium Chelate: Improvements on Fluorescence Energy Transfer," Proc. Nat. Acad. Sci. USA 91, 10024-10028 (1994).
[CrossRef] [PubMed]

S. Stemme, B. Faber, J. Holm, O. Wiklund, J.L. Witztum, G.K. Hansson, "T lymphocytes from human atherosclerotic plaques recognize oxidized Low density lipoprotein," Proc. Nat. Acad. Sci. USA 92, 3893-3897 (1995).
[CrossRef] [PubMed]

Spectrochim. Acta A Mol. Biomol. Spectrosc. (1)

N. Arnaud and J. Georges, "Comprehensive study of the luminescent properties and lifetimes of Eu[3][+] and Tb[3][+] chelated with various ligands in aqueous solutions: influence ofthe synergic agent, the surfactant and the energy level of the ligand triplet," Spectrochim. Acta A Mol. Biomol. Spectrosc. 59, 1829-1840 (2003).
[CrossRef] [PubMed]

Value Health. (1)

W. Castelli, "Lipoproteins and Cardiovascular Disease: Biological Basis and Epidemiological Studies," Value Health. 1, 105-109, (1998).
[CrossRef]

Other (3)

National Cholesterol Education Program. Second report of the expert panel on detection,evaluation, and treatment of high blood cholesterol in adults. NIH Pub. No. 93-3095. Bethesda, MD: National Heart, Lung, and Blood Institute, (1993).

P. R. Selvin, in Applied Fluorescence in Chemistry, Biology and Medicine, edited byW. Rettig, B. Strehmenl, S. Schrader, (Springer Verlag, 1999) pp. 457-487.

N. Sabbatini, S. Perathoner, V. Balzani, B., Alpha, J.M. Lehn, "Antenna Effect in Eu3+ and Tb3+ Cryptates," Supramolecular Photochemistry, Editors: V. Balzani., Reidel, Dordrecht. 187 - 206 (1987).

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Figures (4)

Fig. 1.
Fig. 1.

Absorption spectra of aqueous solutions. O.D. is the optical density (the optical path of the cuvette is 1 mm). a) LDL (7.7 mg/mL) and OxLDL (~ 6 mg/mL);b) EuTc, EuTc:LDL; EuTc:OxLDL.

Fig. 2.
Fig. 2.

a) Fluorescence emission spectra of EuTc and EuTc:LDL aqueous solutions at different lipoprotein concentrations;b) Integrated area (S) below the emission spectra as a function of the LDL concentration. The solid line corresponds to a linear fit to the experimental data.

Fig. 3.
Fig. 3.

a) Fluorescence emission spectra of EuTc and EuTc:OxLDL aqueous solutions at different lipoprotein concentrations;b) Integrated area (S) below the emission spectra as a function of the OxLDL concentration. The solid line corresponds to a linear fit to the experimental data.

Fig. 4.
Fig. 4.

Profile of the Eu emission lifetime in EuTc aqueous solution and in the presence of LDL (500 μL of aqueous solution of EuTc added to 5 μL of LDL at the concentration of 2.84 mg/mL) and OxLDL (500 μL of aqueous solution of EuTc added to 5 μL of OxLDL at the concentration of 2.84 mg/mL). The solid lines are fits of Eq. 1 to the experimental data, whose parameters are given in Table 1.

Tables (1)

Tables Icon

Table 1. Fitting parameters of Eq. 1 to the experimental data shown in Fig. 4.

Equations (2)

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y = y 0 + A 1 e ( t τ 1 ) + A 2 e ( t τ 2 ) ,
τ av = i A i τ i 2 i A i τ i .

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