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A new insight on direct actions of Granulocyte-Colony Stimulating Factor in the myocardium

von Dr. Ana Catarina Ribeiro Carrão

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[1.] Arc/Fragment 044 01 - Diskussion
Zuletzt bearbeitet: 2014-02-26 23:34:10 Hindemith
Arc, Becker et al 1999, Fragment, Gesichtet, KomplettPlagiat, SMWFragment, Schutzlevel sysop

Typus
KomplettPlagiat
Bearbeiter
Hindemith
Gesichtet
Yes.png
Untersuchte Arbeit:
Seite: 44, Zeilen: 1-6
Quelle: Becker et al 1999
Seite(n): H2241, Zeilen: l.col: 21-31
[Intracellular oxidant stress can be monitored by measuring] changes in fluorescence resulting from intracellular probe oxidation. DHE enters the cell and is oxidized by ROS, particularly superoxide, to yield fluorescent ethidium. Ethidium binds to DNA (Eth-DNA), further amplifying its fluorescence. Eth-DNA fluorescence is generally stable but can be decreased by hydroxyl radical attack. Thus increases in DHE oxidation to Eth-DNA (i.e., increases in Eth-DNA fluorescence) are suggestive of superoxide generation. Intracellular oxidant stress was monitored by measuring changes in fluorescence resulting from intracellular probe oxidation. Dihydroethidine (DHE, 1–10 μmol/l; Molecular Probes) enters the cell and is oxidized by ROS, particularly superoxide, to yield fluorescent ethidium. Ethidium binds to DNA (Eth-DNA), further amplifying its fluorescence (5). Eth-DNA fluorescence is generally stable but can be decreased by hydroxyl radical attack (27). Thus increases in DHE oxidation to Eth-DNA (i.e., increases in Eth-DNA fluorescence) are suggestive of superoxide generation.

5. Carter,W. O., P. K. Narayanan, and J. P. Robinson. Intracellular hydrogen peroxide and superoxide anion detection in endothelial cells. J. Leukoc. Biol. 55: 253–258, 1994.

27. Prutz, W. A. Inhibition of DNA-ethidium bromide intercalation due to free radical attack upon DNA. Radiat. Environ. Biophys. 23: 1–18, 1984.

Anmerkungen

The source is not mentioned.

Sichter
(Hindemith) Schumann

[2.] Arc/Fragment 044 15 - Diskussion
Zuletzt bearbeitet: 2014-02-26 10:54:52 Hindemith
Arc, Fragment, Gesichtet, Rocic et al 2007, SMWFragment, Schutzlevel sysop, Verschleierung

Typus
Verschleierung
Bearbeiter
Graf Isolan
Gesichtet
Yes.png
Untersuchte Arbeit:
Seite: 44, Zeilen: 15-25
Quelle: Rocic et al 2007
Seite(n): H2730, Zeilen: left col. 33-47
For the in vitro studies in freshly isolated cardiomyocytes (Cardiomyocyte Isolation for details), G-CSF was administered for 10 min (total treatment time 30 min), and DHE was added during the last 20 min of treatment with G-CSF. Cells were then immediately observed under a fluorescent microscope. For in vivo studies, DHE was injected into the LV (60 μg/kg, n=3/group) for 20 min before two consecutive periods of ischemia-reperfusion (40s occlusion followed by 20min reperfusion and another 40s occlusion and 20min reperfusion). Animals were then sacrificed. The heart was removed, frozen in optimum cutting temperature compound on dry ice, and stored at -80°C until sectioning. Sections (5 μm) were made in a cryomicrotome and were mounted on glass slides. DHE fluorescence was detected with excitation/emission at 518/605 nm. All images were analyzed at the same microscope settings, and fluorescence intensity was obtained by Metamorph Software on three hearts (10 sections per heart). For HCAEC cell culture, DHE was administered during the last 20 min of treatment. Cells were then observed immediately under a fluorescent microscope. For in vivo studies, DHE or DCF were injected into the LV (60 μg/kg) for 20 min before two consecutive periods of ischemia-reperfusion (40 s occlusion followed by 20 min reperfusion and another 40 s occlusion and 20 min reperfusion). Animals were then killed. The heart was removed, frozen in optimum cutting temperature compound on dry ice, and stored at -70°C until sectioning. Sections (5 μm) were made in a cryomicrotome and were mounted on glass slides. DHE or DCF fluorescence was detected with excitation/emission at 518/605 nm (for DHE) or 480/515 nm (for DCF). All images were analyzed at the same microscope settings, and relative fluorescence readings were obtained by Metamorph Software on three hearts (5 consecutive sections per heart).
Anmerkungen

Though taken verbatim nothing has been marked as a citation and the source is not given.

Sichter
(Graf Isolan), Hindemith


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Letzte Bearbeitung dieser Seite: durch Benutzer:Hindemith, Zeitstempel: 20140226233500

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