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Angaben zur Quelle [Bearbeiten]

Autor     Eiji Toyota, David C. Warltier, Tommy Brock, Erik Ritman, Christopher Kolz, Peter O'Malley, Petra Rocic, Marta Focardi, William M. Chilian
Titel    Vascular Endothelial Growth Factor Is Required for Coronary Collateral Growth in the Rat
Zeitschrift    Circulation
Herausgeber    American Heart Association
Ausgabe    112
Jahr    2005
Seiten    2108-2113
ISSN    1524-4539
DOI    10.1161/CIRCULATIONAHA.104.526954
URL    http://circ.ahajournals.org/content/112/14/2108.full.pdf

Literaturverz.   

yes
Fußnoten    yes
Fragmente    7


Fragmente der Quelle:
[1.] Arc/Fragment 040 03 - Diskussion
Zuletzt bearbeitet: 2014-02-26 23:06:53 Guckar
Arc, BauernOpfer, Fragment, Gesichtet, SMWFragment, Schutzlevel sysop, Toyota et al 2005

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Quelle: Toyota et al 2005
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Male Sprague-Dawley rats (3–4 mo old, 300–350 g) were used for chronic (5 days) implantation of a pneumatic occluder over the left anterior descending coronary artery (LAD; as described by Toyota et al.96) to produce repetitive ischemia (RI). Briefly, for surgery, rats were premedicated (ketamine 50 mg/mL plus acepromazine 2.5 mg/mL plus torbuterol 2.0 mg/mL, 0.2 mL/100 g body weight, i.p.) and intubated. Oral intubation (I6-G polyethylene tubing) was done under direct observation of the vocal cords with an otoscope. General anesthesia was introduced and maintained by sevoflurane inhalation (1.0% to 2.0%, with 100% oxygen). Body temperature was controlled at 37°C by an electric heating table. Surgery was performed using aseptic technique. The animal was initially placed on its dorsal side, and after a neck incision, the right carotid artery was isolated, and a PE-50 catheter filled with heparin (10 U/mL)-saline was inserted. This tubing was used for monitoring of systemic hemodynamics, sampling arterial blood, and maintaining the blood volume. Blood pH, PaO2, PaCO2, and systemic hemodynamics were maintained within physiological ranges throughout the surgery. The animal was repositioned on its right side, and the heart was exposed by left thoracotomy. A mini-pneumatic snare occluder (see the Mini- Pneumatic Snare Occluder section for details) was implanted around the mid to proximal LAD. Confirmation that the occluder was functional, i.e., producing myocardial ischemia, was determined initially by observation of blanching and hypokinesis of the left ventricle (LV) during inflation. Rats were randomly divided into 3 groups based on the type of measurement: coronary blood flow (CBF) (neutron activated microspheres), oxidative stress analysis (Dihydroethidium, DHE) or vascular imaging (micro-CT). CBF was measured during coronary occlusion to determine flows to the normal and collateral-dependent zones (see the Microsphere measurements of myocardial and collateral-dependent blood flow and the Coronary Microvascular imaging with Cryomicrotome sections). After instrumentation and measurements, the chest was closed under positive end-expiratory pressure, and the thoracic cavity was evacuated of air. The occluder was tunnelled subcutaneously and exteriorized between the scapulae. This catheter was protected by a stainless steel spring coil connected to a ring that was secured subcutaneously between the scapulae.

96. Toyota E, Warltier DC, Brock T, Ritman E, Kolz C, O'Malley P, Rocic P, Focardi M, Chilian WM. Vascular endothelial growth factor is required for coronary collateral growth in the rat. Circulation. 2005;112:2108-2113.

Male Wistar rats (290- to 360-g body weight; n=50) were used for experiments (preparations were successful in 39 animals). For surgery, rats were premedicated (ketamine 50 mg/mL plus acepromazine 2.5 mg/mL plus torbuterol 2.0 mg/mL, 0.2 mL/100 g body weight IP) and intubated. Oral intubation (I6-G polyethylene tubing) was done under direct observation of the vocal cords with an otoscope. General anesthesia was introduced and maintained by sevoflurane inhalation (1.0% to 2.0%, with 100% oxygen). Body temperature was controlled at 37°C by an electric heating table. Surgery was performed using aseptic technique. The animal was initially placed on its dorsal side, and after a neck incision, the right carotid artery was isolated, and a PE-50 catheter filled with heparin (10 U/mL)-saline was inserted. This tubing was used for monitoring of systemic hemodynamics, sampling arterial blood, and maintaining the blood volume. Blood pH, PaO2, PaCO2, and systemic hemodynamics were maintained within physiological ranges throughout the surgery. The animal was repositioned on its right side, and the heart was exposed by left thoracotomy. A mini-pneumatic snare occluder (see the Mini-Pneumatic Snare Occluder section for details) was implanted around the mid to proximal left anterior descending coronary artery (LAD). Confirmation that the occluder was functional, ie, producing myocardial ischemia, was determined initially by observation of blanching and hypokinesis of the left ventricle (LV) during inflation. Rats were randomly divided into 2 groups based on the type of measurement: coronary blood flow (CBF) (radioactive microspheres, n=20) or vascular imaging (micro-CT, n=13). CBF was measured during coronary occlusion to determine flows to the normal and collateral-dependent zones (see the Measurement of Coronary Blood Flow and the Coronary Vascular Imaging With Micro-CT sections). After instrumentation and measurements, the chest was closed under positive end-expiratory pressure, and the thoracic cavity was evacuated of air. An intraperitoneal catheter (PE-50) for drug administration was inserted, and this catheter and the occluder were tunneled subcutaneously and exteriorized between the scapulae. These catheters were protected by a stainless steel spring coil connected to a ring that was secured subcutaneously between the scapulae.
Anmerkungen

The source is given at the beginning of the section in a way that indicates that the here described method is similar to the one employed by the authors of the source. However, it is not clear to the reader that the description of the method is a largely literal copy of the source.

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(Hindemith) Schumann

[2.] Arc/Fragment 041 01 - Diskussion
Zuletzt bearbeitet: 2014-02-26 23:07:05 Guckar
Arc, Fragment, Gesichtet, SMWFragment, Schutzlevel sysop, Toyota et al 2005, Verschleierung

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Untersuchte Arbeit:
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Quelle: Toyota et al 2005
Seite(n): 2109, Zeilen: l.col: 46ff
[After the surgery, analgesic] (buprenorphine 0.05 mg/kg s.c.) and antibiotic (enrofloxacin 10 mg/kg s.c.) compounds were administered. Rats were observed in a recovery cage for 2 hours and then transferred to the animal care facility. For 3 days after the surgery, buprenorphine (0.5 mg/kg BID mixed in strawberry Jello) was taken orally for pain relief. On the fourth day after the surgery, the ischemic protocol was started (see the Experimental Protocol section). After 5 days of the experimental protocol, the rats were anesthetized and the chest was opened by mid thoracotomy. In CBF measurement or DHE analysis groups, the hearts were excised at the end of procedures, and the tissue was prepared for analyses.

3.2 Mini-Pneumatic Snare Occluder for Rat Heart

A mini-pneumatic snare occluder consisting of a mini-balloon, sheath tubing, suture, and catheter (Fig. 11) was placed on the LAD of the rat heart. The balloon (7 mm long) is made of soft latex membrane and is sufficiently pliable to give negligible physical force on the coronary vessels during balloon deflation. The balloon is mounted within an umbrella sheath (3.2 or 4.8 mm in diameter, 12 mm in length; protects the balloon from fibrous infiltration). Prolene (5– 0) is passed around the LAD and attached to the sheath, securing the occluder to the heart, so that myocardial ischemia is produced by balloon inflation. Inflation volume is small (0.2 to 0.25 mL air), but occlusion occurs by 2 physical actions: “crimping” the LAD toward upward/outside and compressing the LAD by the inflated balloon/sheath. The balloon is connected to a catheter (PE-50) that is exteriorized. Balloon inflation and deflation are controlled from outside the rat cage.

41a diss.png

Fig. 11 - Schematic diagram of the mini-pneumatic snare and its actions. Top: Cross-sectional and longitudinal views when the balloon is deflated. Bottom: Views during inflation. The artery is patent when the balloon is deflated, but during inflation, a snare situated underneath the artery is pulled “upward” during inflation, producing the coronary occlusion94.


94. Kappel A, Ronicke V, Damert A, Flamme I, Risau W, Breier G. Identification of Vascular Endothelial Growth Factor (VEGF) Receptor-2 (Flk-1) Promoter/Enhancer Sequences Sufficient for Angioblast and Endothelial Cell- Specific Transcription in Transgenic Mice. Blood. 1999;93:4284-4292.

After the surgery, analgesic (buprenorphine 0.05 mg/kg SC) and antibiotic (enrofloxacin 10 mg/kg SC) were administered. Rats were observed in a recovery cage for 2 hours and then transferred to the animal care facility. For 3 days after the surgery, buprenorphine (0.5 mg/kg BID mixed in strawberry Jello) was taken orally for pain. On the fourth day after the surgery, ischemic protocol was started (see the Experimental Protocol section).

After 10 days of the experimental protocol, the rats were anesthetized, and the chest was opened by mid thoracotomy. In the micro-CT group, the hearts were immediately excised. In CBF measurement group, blood flow to the normal and collateraldependent zones during coronary occlusion was measured. The heart was excised at the end of measurements, and the tissue was prepared for analyses of radioactivity.

Mini-Pneumatic Snare Occluder for Rat Heart

We developed a mini-pneumatic snare occluder (patent application serial number: 11/071,617, E.T. and W.M.C.) consisting of a mini-balloon, sheath tubing, suture, and catheter (Figure 1). The balloon (7 mm long) is made of soft latex membrane and is sufficiently pliable to give negligible physical force on the coronary vessels during balloon deflation. The balloon is mounted within an umbrella sheath (3.2 or 4.8 mm in diameter, 12 mm in length; protects the balloon from fibrous infiltration). Prolene (5– 0) is passed around the LAD and attached to the sheath, securing the occluder to the heart, so that myocardial ischemia is produced by balloon inflation. Inflation volume is small (0.2 to 0.25 mL air), but occlusion occurs by 2 physical actions: “crimping” the LAD toward upward/outside and compressing the LAD by the inflated balloon/sheath. The balloon is connected to a catheter (PE-50) that is exteriorized. Balloon inflation and deflation are controlled from outside the rat cage.

41a source.png

Figure 1. Schematic of the mini-pneumatic snare and its actions. Top, Cross-sectional and longitudinal views when the balloon is deflated. Bottom, Views during inflation. The artery is patent when the balloon is deflated, but during inflation, a snare situated underneath the artery is pulled “upward” during inflation, producing the coronary occlusion.

Anmerkungen

At the beginning of the previous page the source is mentioned, but without indication that the following two pages are taken from it.

The given source Kappel et al. (1999) doesn't contain any of the copied material.

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(Hindemith) Schumann

[3.] Arc/Fragment 042 16 - Diskussion
Zuletzt bearbeitet: 2014-02-26 23:07:17 Guckar
Arc, Fragment, Gesichtet, SMWFragment, Schutzlevel sysop, Toyota et al 2005, Verschleierung

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Quelle: Toyota et al 2005
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CBF was measured with neutron activated microspheres (Biopal, 15 μm) before initiation of (after all the instruments had been implanted at the time of the initial surgery) and at the end of the repetitive occlusions (when the rats were anesthetized and the chest was open to mimic the conditions of the first measurement) to measure normal zone flows and flow in the developed collaterals. For the first measurement, neutron activated microspheres labelled with Samarium were mixed with fluorescent (FITC) microspheres (Invitrogen, 10 μm) to identify the collateral-dependent region as described below. For the second measurement, Gold labelled microspheres were used. The microspheres (5×105) were injected directly into the LV cavity via the LV apex during LAD occlusion with a 29-gauge insulin syringe over a 10-second period. During the course of the procedures, systemic pressure and heart rate were recorded (386- BIOS, American Megatrends Inc). The heart was excised and fresh LV was sliced along the short axis and observed with a dissecting microscope and fluorescent light source. The collateral-dependent area (ischemic zone, LAD region) was distinguished as the [area without fluorescent microspheres.] CBF was measured with radioactive microspheres (Perkins Elmer; φ; 15 μm; 95Nb and 103Ru, ≈4.5x105) before initiation of (after all the instruments had been implanted at the time of the initial surgery) and at the end of the repetitive occlusions (when the rats were anesthetized and the chest was open to mimic the conditions of the first measurement) to measure normal zone flows, native collateral flow, and flow in the developed collaterals. For the first measurement, radioactive microspheres were mixed with fluorescent (FITC) microspheres (φ, 10 μm; ≈9x105, Fluoresbrite Yellow Green, Polysciences, Inc) to identify the collateral-dependent region as described below. For the second measurement, the other nuclidelabeled microspheres were used. The microspheres were agitated for 15 minutes, suspended in saline (total volume, 150 μL), and then injected directly into the LV cavity via the LV apex during LAD occlusion with a 29-gauge insulin syringe over a 10-second period. [...] During the course of the procedures, systemic pressure and heart rate were recorded (386- BIOS, American Megatrends Inc).

The heart was excised and fixed in 4% paraformaldehyde solution overnight. The fixed LV was sliced along the short axis and observed with a dissecting microscope and fluorescent light source (LT-9800, Lightools Research). The collateral-dependent area (LAD region) was distinguished as the area without fluorescent microspheres.

Anmerkungen

The source is not given.

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[4.] Arc/Fragment 043 01 - Diskussion
Zuletzt bearbeitet: 2014-02-27 19:44:56 WiseWoman
Arc, Fragment, Gesichtet, SMWFragment, Schutzlevel sysop, Toyota et al 2005, Verschleierung

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The control area (non-LAD LV region) was determined by the area of distribution of the fluorescent microspheres. The normal and collateral-dependent zones were divided with a blade, and for each the total weight was measured. Collateral flow was calculated as a ratio between activity (dpm/g) of the tissue samples from the LAD-dependent and normal zones. The control area (non-LAD LV region) was determined by the area distribution of the fluorescent microspheres. The normal and collateral-dependent zones were divided with a blade, and each total weight was measured. CBF (mL · min-1 · g-1) in each area was calculated from the following formula: CBF=[(radioactive counts in myocardial specimen)x(blood withdrawal rate)/(radioactive count in blood)]/(weight of myocardial specimen).
Anmerkungen

The copied text begins on the previous page: Arc/Fragment_042_16

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[5.] Arc/Fragment 049 02 - Diskussion
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Arc, Fragment, Gesichtet, SMWFragment, Schutzlevel sysop, Toyota et al 2005, Verschleierung

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Evaluation of collateral growth was performed after completion of the protocol by a simple functional test: total coronary occlusion. The rationale for this procedure was that if collaterals were developed, then occlusion would not induce functional disturbances. Alternatively, if collaterals were not mature, then occlusion would cause hemodynamic disturbances. After the second measurement of coronary blood flow, the coronary occlusion was maintained, systemic hemodynamics were measured and also the number of arrhythmias. In animals without collaterals, coronary occlusion caused deterioration of systemic hemodynamics and arrhythmias, including premature ventricular contractions, ventricular tachycardia, and ventricular fibrillation; in animals with well developed collaterals, no such adverse effects were noted. We evaluated collateral growth after completion of the protocol by a simple functional test: total coronary occlusion. The rationale for this

[page 2110]

procedure was that if collaterals were developed, then occlusion would not induce functional disturbances. Alternatively, if collaterals were not mature, then occlusion would cause hemodynamic disturbances. After the second measurement of coronary blood flow, we maintained the coronary occlusion (n=9 in control group, n=6 in anti-VEGF group) and measured systemic hemodynamics and the number of arrhythmias. In animals without collaterals, coronary occlusion caused deterioration of systemic hemodynamics and arrhythmias, including premature ventricular contractions, ventricular tachycardia, and ventricular fibrillation; in animals with well-developed collaterals, no such adverse effects were noted.

Anmerkungen

The source is not mentioned here.

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[6.] Arc/Fragment 049 14 - Diskussion
Zuletzt bearbeitet: 2014-02-26 23:07:35 Guckar
Arc, Fragment, Gesichtet, SMWFragment, Schutzlevel sysop, Toyota et al 2005, Verschleierung

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Quelle: Toyota et al 2005
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After the completion of treatments, the heart was removed, and the left ventricle (LV) was sliced along the short axis and observed with a dissecting microscope under fluorescent light. The collateral-dependent area (LAD region, ischemic zone) was distinguished as the area without fluorescent microspheres. The control area (non-LAD LV region, normal zone) was determined by the area of distribution of the fluorescent microspheres (Fig. 13). The heart was excised and fixed in 4% paraformaldehyde solution overnight. The fixed LV was sliced along the short axis and observed with a dissecting microscope and fluorescent light source (LT-9800, Lightools Research). The collateral-dependent area (LAD region) was distinguished as the area without fluorescent microspheres. The control area (non-LAD LV region) was determined by the area distribution of the fluorescent microspheres.
Anmerkungen

The source is not given. To be continued on the next page: Arc/Fragment_050_01

Note that a similar text can also be found further up in the thesis, c.f. Arc/Fragment_042_16, Arc/Fragment_043_01

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[7.] Arc/Fragment 050 01 - Diskussion
Zuletzt bearbeitet: 2014-02-26 23:07:44 Guckar
Arc, Fragment, Gesichtet, KomplettPlagiat, SMWFragment, Schutzlevel sysop, Toyota et al 2005

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The normal and collateral-dependent zones were divided with a blade, and each total weight was measured. The normal and collateral-dependent zones were divided with a blade, and each total weight was measured.
Anmerkungen

The copied text starts on the previous page: Arc/Fragment_049_14

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