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green were counted as dead cells. The cells were counted by examining 10 random microscopy fields and identiffing the apoptotic and live myocytes. Data are expressed as a count of apoptotic cells/l00 cells. II.e.Oxidative stress studies Oxidative stress was quantified using a previously described method (Kajstura et al. 2001). The endogenous production of reactive 02 species was quantified utilizing a 5- (6)-chloromethyl-2'7'-dihydroflourescein diacetate probe (CM-H2 DCFDA) (Molecular Probes, Eugene, Oregon, USA). 100 pM stock solution of CM-H2 DCFDA in DMSO v/as prepared fresh. Cell loading with fluorescent probes was achieved by incubating the cells with 10 ¡rM solution of CM-H2 DCFDA in PBS for 30 minutes. lncubation was performed in a humidified chamber, protected from light, at 37oC. Fluorescence intensity, which was proportional to the level of oxidative stress, was rapidly measured by a fluorescent microscopy. An excitation wavelength of 485 nm and emission wavelength of 530 nm were used. The analysis of 10-12 randomly chosen fieids of microscopy was performed using the Olympus BX 51 fluorescent microscope equipped with green filter, 10x0.30 lens, "cool snap" digital camera (Photometrics, Tucson, Arizorra, USA) and digital imaging processing software (Image Pro Plus, Media Cybernetics, Silver Springs, Maryland, USA). 76
III. Protein Assessment and Statistical Analvsis la order to equalize protein loading, protein concentration was analyzed using the modified Lowry method (LOWRY et al. 1951). Ponceau S stain, Coomasie blue, B- actin (Sigma- Aldrich, Oakville, Ontario, Canada) and troponin C IgG antibodies (Cell Signaling, Beverly, MA, USA) were used to ensure equal loading of the samples. Data were expressed as Mean + SEM. For statistical analysis of the data, for each group the mean was compared to its control thus one way ANOVA and Bonferroni's test was used to identiff differences between the control and each experimental group. Statisiical difference with P
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il\VOLVEMENT OF RETII\OIC ACID II{
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Title: AKNOWLEDGMENTS DEDICATION LI
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IV.b.3. The uptake to peripheral ti
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II.a. Total RAR and total RXR recep
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ACKNO\ryLEDGMENTS I must say that m
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DEDICATION To my toughest critic, m
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Figure: LIST OF FIGURES Page: 1. Ch
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30. A and B The expression of anti-
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treated (ADR), trolox treated (TROL
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INTRODUCTION Adriamycin, an anthrac
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apoptosis, which will ultimately pr
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LITERATURE REVIE\il I. Adriamycin i
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Binding and alkylation of DNA Inter
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myocardial inflammation which was s
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1990; Kusuoka et al. l99l; Olson et
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levels of antioxidant enzymes (Odom
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causing the increased leakage of el
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mechanisms which are involved in th
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to protect against adriamycin-induc
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The usage of Probucol.' Most promis
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II. Oxidative stress II.a. Introduc
Page 43 and 44: of a large array of cellular abnorm
Page 45 and 46: menstruation (Kokawa et al. 1996),
Page 47 and 48: DNA(Cohen 1997; Saikumar et al. 199
Page 49 and 50: pathogenesis of heart failure. A nu
Page 51 and 52: elonging to spontaneous hlpertensiv
Page 53 and 54: AlþTrsns Retlnolc Acld fAfRAl g€
Page 55 and 56: dehydrogenase (SDR), alcohol dehydr
Page 57 and 58: (Bavik et a|. 1991). Soon after its
Page 59 and 60: et al. 1994; Napoli 1996). The func
Page 61: is achieved through its binding to
Page 64 and 65: Retinoic acid signaling pathwaybegi
Page 66 and 67: (Bavik et al. 1997). One of the mos
Page 68 and 69: cells (Gaetano et ai. ZOot¡. The s
Page 70 and 71: neuroblastoma, genn cell tumors and
Page 72 and 73: presence of two distinctive pathway
Page 74 and 75: cytochrome C release from mitochond
Page 76 and 77: peroxidation (Sultana et a|.2004).
Page 78 and 79: adipose tissue and to a lesser exte
Page 80 and 81: such as; increased generation of re
Page 82 and 83: physiological and pathological proc
Page 84 and 85: Protective effects of PPAR y agonis
Page 86 and 87: IIYPOTHESIS This study tests the hy
Page 88 and 89: I.c.Collection of Tissues Hearts we
Page 90 and 91: thoracotomy was performed and heart
Page 92 and 93: Il.c.Western Blot Analvsis Western
Page 96 and 97: RESULTS I.In Vivo Studies I.a.Gener
Page 101: Lc. Retinoic acid receptors Three w
Page 108: The RAR o and PPAR-õ receptors RNA
Page 113 and 114: significantly decreased when compar
Page 116: IL b.7. RAR alPha recePtor levels T
Page 120: ILb. 3. RAR sammø recEtor levels F
Page 124: ILb. 5. RXR baa recEúor levels The
Page 129: visible staining were accounted as
Page 137 and 138: DISSCUSION I. Adriamvcin CardiomvoP
Page 139 and 140: Treatment with adriamycin has been
Page 141 and 142: High RA 1 Adriamycin I oxidative st
Page 143 and 144: study as well as in others (Kumar e
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It is reported that PPAR y receptor
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expression. These effects of trolox
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REFERENCES Abdul Hamied,T.A., D.Par
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Bashor,M.M., D.O.Toft, and F.Chytll
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distribution of PPAR-alpha, -beta,
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colbert,M.C., D.G.Hall, T.R.Kimball
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Diep,Q.N., M'El Mabrouk, J.S.Cohn,
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Evans,R.M. 1988. "The steroid and t
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Ghione M. Development of Adriamycin
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Herman,E., R.Young, and S.Krop . Ig
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morphogenetic protein-2 inhibits se
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Khandoudi,N., P.Delerive, I.Berrebi
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Kusuoka,H., S.Futaki, Y.Koretsune,
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and myosin heavy chain gene express
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Mehta,J.L., B.Hu, J.Chen, and D.Li.
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Morriss-Kay,G.M. and s.J.ward. 1999
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Notario,B., M.Zamora, O.Vinas, and
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petkovich,M., N.J.Brand, A.Krust, a
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Ruberte,E., P.Dolle, P.Chambon, and
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Sharov,V.G., H.N.Sabbah, H.Shimoyam
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spiegelman,B.M. and J.s.Flier .1996
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Teebor,G.w., R.J.Boorstein, and J.c
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vanVeen,T.A.,H.V.vanRijen,R.F.V/ieg
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wi1liams,J.B. and J.L.Napoli. 1985.
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peroxisome proliferator-activated r
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