Post on 05-Feb-2018
HASIL PENELITIAN
PENGARUH EKSTRAK JINTAN HITAM (Nigella Sativa) TOPIKAL
TETES MATA TERHADAP PENURUNAN EKSPRESI Nuclear Factor
Kappa Beta (NF-Kβ) PADA KORNEA TIKUS JANTAN (Rattus Novergicus
Strain Wistar) MODEL INFLAMASI
Oleh:
IMANIYAH HUSNI
201110330311151
UNIVERSITAS MUHAMMADIYAH MALANG
FAKULTAS KEDOKTERAN
2015
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KATA PENGANTAR
Puji syukur kehadirat Allah SWT atas rahmat dan inayah-Nya sehingga
penulis diberi kekuatan dan kemampuan dalam menyelesaikan karya tulis akhir
ini yang berjudul “PENGARUH EKSTRAK JINTAN HITAM (Nigella Sativa)
TOPIKAL TETES MATA TERHADAP PENURUNAN EKSPRESI Nuclear
Factor Kappa Beta (NF-Kβ) PADA KORNEA TIKUS JANTAN (Rattus
Novergicus Strain Wistar) MODEL INFLAMASI”. Karya tulis akhir ini diajukan
untuk memenuhi persyaratan Pendidikan Sarjana Fakultas Kedokteran Universitas
Muhammadiyah Malang.
Pada kesempatan ini, penulis ingin menyampaikan terimakasih kepada :
1. dr. Irma Suswati, M.Kes beserta para pembantu dekan selaku Dewan
Dekanat Fakultas Kedokteran UMM.
2. dr. Bragastio Sidharta, Sp.M, selaku pembimbing 1 atas kesabaran,
kebaikan hati serta kesediaan dalam meluangkan waktu dalam membantu,
membimbing dan mengarahkan penulis dalam menyelesaikan karya tulis
akhir ini.
3. dr. Thontowi Djauhari, NS, M.Kes, selaku pembimbing 2 atas kesabaran,
kebaikan hati serta kesediaan dalam meluangkan waktu dalam membimbing
penulis hingga dapat menyelesaikan menyelesaikan dengan baik.
4. dr. Heru Priyo Husodo, Sp.Rad, selaku penguji tugas akhir ini yang telah
membantu dan memberikan masukan kepada penulis dalam menyelesaikan
karya tulis akhir ini.
5. Seluruh dosen beserta karyawan-karyawati Fakultas Kedokteran UMM yang
selalu membantu dan membimbing penulis.
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6. Kedua orang tua penulis, Ayahanda Husni Husain Hasan dan Ibunda
Djasna Ali atas doa dan dukungannya yang tak pernah putus. Kedua saudari
penulis Inayati Husni dan Amaliyah Husni atas bantuan doa, motivasi dan
tenaganya beserta seluruh keluarga tercinta yang senantiasa setia
mendukung penulis dalam menyelesaikan karya tulis akhir ini.
7. Seluruh staf TU dan laboran, Mas Miftah dan Pak Joko yang bersedia
membantu setiap tahapan proses penelitian hingga penyelesaian tugas akhir
ini.
8. Sejawat FK UMM angkatan 2011 “Adorables” atas dukungan, bantuan, dan
kerjasamanya. Semoga kelak dapat menjadi dokter-dokter profesional yang
dapat bermanfaat bagi orang lain serta dapat menjalankan amanah sebagai
dokter karena Allah SWT.
9. Sahabat-sahabat terbaik yang selalu membantu selama penyelesaian karya
tulis akhir ini, Mariyah,Lilly, Tari, Alfi, Laksita, Putrinda, Kinanthi, Jeanny,
Ilma, Dwi, Friska, Hakimah, terimakasih atas doa, dukungan dan bantuan
yang telah diberikan selama ini.
10. Kelompok Jintan Hitam Lilly, Muqsit, Rofwiun atas dukungan dan
kerjasama dalam penyelesaian karya tulis akhir ini.
11. Keluarga besar Skill laboratorium FK UMM atas doa dan dukungannya.
12. Keluarga Besar Al-Zahrawi FK UMM atas doa, dukungan serta bantuan
yang diberikan selama ini.
13. Semua pihak yang terlibat secara langsung maupun tidak langsung,
terimakasih atas bantuan dan dukungannya.
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Penulis menyadari bahwa tugas akhir ini masih jauh dari kesempurnaan.
Oleh karena itu, penulis sangat mengharapkan saran, kritik dan masukan yang
membangun. Dengan mengharapkan Keridhaan-Nya, semoga karya tulis ini dapat
menambah wawasan keilmuan dan bermanfaat bagi semua pihak.
Malang, Agustus 2015
Penulis
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DAFTAR ISI
Halaman
LEMBAR PENGESEHAN .....................................................................................i
LEMBAR PENGUJIAN .........................................................................................ii
KATA PENGANTAR ............................................................................................iii
ABSTRAK ..............................................................................................................vi
DAFTAR ISI ........................................................................................................viii
DAFTAR GAMBAR .............................................................................................xi
DAFTAR TABEL .................................................................................................xii
DAFTAR SINGKATAN ......................................................................................xiii
DAFTAR LAMPIRNAN ......................................................................................xiv
BAB 1 PENDAHULUAN
1.1 Latar Belakang .................................................................................................1
1.2 Rumusan Masalah .............................................................................................4
1.3 Tujuan Penelitian ..............................................................................................4
1.3.1 Tujuan Umum .........................................................................................4
1.3.2 Tujuan Khusus ........................................................................................4
1.4 Manfaat Penelitian ............................................................................................5
1.4.1 Akademis .................................................................................................5
1.4.2 Klinis .......................................................................................................5
BAB 2 TINJAUAN PUSTAKA
2.1 Kornea ...............................................................................................................6
2.1.1 Definisi ...................................................................................................6
2.1.2 Anatomi Kornea ......................................................................................7
2.2 Inflamasi .........................................................................................................10
2.2.1 Definisi ..................................................................................................10
2.2.2 Proses Inflamasi ....................................................................................11
2.3 Inflamasi Kornea ............................................................................................13
2.4 Jintan Hitam (Nigella sativa) ..........................................................................14
2.4.1 Definisi dan Taksonomi .........................................................................14
2.4.2 Komposisi Kimia Nigella sativa ...........................................................15
2.5 Thymoquinon ..................................................................................................17
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2.5.1 Definisi .................................................................................................17
2.5.2 Mekanisme Thymoquinon menghambat aktifasi
Nuclear Factor Kappa Beta (NF-Kβ) ...................................................18
2.6 Nuclear Factor Kappa Beta (NF-Kβ) ............................................................19
2.6.1 Definisi ..................................................................................................19
2.6.2 Struktur Nuclear Factor Kappa Beta (NF-Kβ) .....................................21
2.6.3 Mekanisme Nuclear Factor Kappa Beta (NF-Kβ) ...............................22
2.6.5 Aktivasi Nuclear Factor Kappa Beta (NF-Kβ) ....................................23
2.6.6 Jalur Aktivasi Nuclear Factor Kappa Beta (NF-Kβ) ............................24
BAB 3 KERANGKA KONSEP DAN HIPOTESIS
3.1 Kerangka Konsep ............................................................................................27
3.2 Hipotesis .........................................................................................................28
BAB 4 METODOLOGI PENELITIAN
4.1 Rancangan Penelitian .....................................................................................29
4.2 Tempat dan Waktu Penelitian ........................................................................29
4.3 Populasi dan Sampel .......................................................................................29
4.3.1 Populasi .................................................................................................29
4.3.2 Sampel ...................................................................................................29
4.3.3 Besar Sampel Penelitian .......................................................................30
4.3.4 Teknik Pengambilan Sampel ................................................................32
4.3.5 Karakteristik Sampel Penelitian ............................................................32
4.3.6 Variabel Penelitian ................................................................................33
4.3.7 Definisi Operasional Variabel Penelitian ..............................................33
4.4 Alat dan Bahan ...............................................................................................34
4.3.1 Alat .......................................................................................................34
4.3.2 Bahan ...................................................................................................34
4.5 Kerangka Operasional Penelitian ...................................................................35
4.6 Prosedur Penelitian ........................................................................................36
4.6.1 Adaptasi ................................................................................................36
4.6.2 Pembuatan Ekstrak Nigella sativa ........................................................36
4.6.3 Pembuatan Solusio Nigella sativa Tetes Mata Topikal .........................37
4.6.4 Perlakuan Sampel ..................................................................................37
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4.6.5 Pengambilan Sampel Kornea untuk Pengamatan
Ekspresi NF-Kβ .....................................................................................38
4.6.6 Pemeriksaan NF-Kβ dalam Metode Imunohistokimia .........................39
4.6.7 Pengamatan Ekpresi NF-Kβ dengan
Metode Imunhistokimia pada Kornea ...................................................40
4.6.8 Penghitungan Ekspresi NF-Kβ ..............................................................41
4.7 Analisa Data ...................................................................................................42
BAB 5 HASIL PENELITIAN DAN ANALISIS DATA
5.1 Hasil Penelitian ...............................................................................................37
5.2 Analisis Data ....................................................................................................38
5.2.1 Uji Kruskal Wallis ..................................................................................38
5.2.2 Uji Post-Hoc (Mann-Whitney) ...............................................................39
5.2.3 Uji Korelasi …………..........................................................................39
BAB 6 PEMBAHASAN ........................................................................................41
BAB 7 KESIMPULAN DAN SARAN
7.1 Kesimpulan ......................................................................................................52
7.2 Saran ................................................................................................................52
DAFTAR PUSTAKA ............................................................................................53
LAMPIRAN ...................... ...................................................................................59
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DAFTAR GAMBAR
Gambar Halaman
2.1 Anatomi Mata ...................................................................................................7
2.2 Anatomi Kornea ..............................................................................................9
2.3 Jintan Hitam (a) bunga dan (b) biji .................................................................14
2.4 Struktur Kimia Thymoquinon .........................................................................17
2.5 Mekanisme Thymoquinon Menghambat Aktifasi
Nuclear Factor Kappa Beta (NF-Kβ) ...........................................................18
2.6 Protein Nuclear Factor Kappa Beta (NF-Kβ) ................................................20
2.7 Kelas Nuclear Factor Kappa Beta (NF-Kβ) .................................................21
2.8 Mekanisme Nuclear Factor Kappa Beta (NF-Kβ) .........................................22
2.9 Jalur Aktifasi Nuclear Factor Kappa Beta (NF-Kβ) ......................................25
3.1 Kerangka Konsep ...........................................................................................26
4.1 Kerangka Operasional Penelitian ...................................................................34
4.2 Lokasi Pengambilan Sampel Kornea ..............................................................38
5.1 Gambaran ekspresi NF-Kβ ............................................................................44
5.2 Ekspresi NF-Kβ ..............................................................................................45
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DAFTAR TABEL
Tabel Halaman
2.1 Komposisi Kimia Nigella sativa .....................................................................16
2.2 Komposisi Minyak Nigella sativa ...................................................................16
4.1 Klasifikasi perhitungan NF-Kβ ........................................................................40
4.2 Jadwal Penelitian ............................................................................................41
5.1 Hasil Penelitian Ekspresi NF-Kβ pada Kornea
Mata Tikus (Rattus novergius strain wistar) Jantan .......................................44
5.2 Hasil Kruskal Wallis .......................................................................................46
5.3 Hasil Uji Post-Hoc (Mann-Whitney) ...............................................................46
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DAFTAR SINGKATAN
a-MSH : Alpha-Melanosit Stimulating Hormon
BAFF : B Cell Activating Factor
DBD : DNA-Binding Domain
DNA : Deoxyribonucleic Acid
DTQ : Dimmer Thymoquinone
IKK : IKB Kinase
NF-Kβ : Nuclear Factor Kappa Betta
NIK : NFkβ-Inducing Kinase
NLS : Localization Signals
NSO : Nigella sativa Oil
TGF : Transformidezzing beta Growth Factor
THQ : Thymohydroquinone
THY : Timol
TLR : Toll-Like Receptor
TNF : Tumor Necrosis Factor
TNFR : Tumor Necrosis Factor Receptor
TQ : Thymoquinone
RANK : Receptor Activator of Nuclear Factor Kb
RE : Response Elements
RHD : Rel Homology Domain
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DAFTAR LAMPIRAN
Halaman
Lampiran 1 : Data Hasil Penelitian ......................................................................59
Lampiran 2 : Analisis Data ..................................................................................60
Lampiran 3 : Dokumentasi Kegiatan ....................................................................66
Lampiran 4 : Surat Determinasi Tanaman ............................................................70
Lampiran 5 : Surat Keterangan Ekstrak Jintan Hitam .........................................71
Lampiran 6 : Surat Keterangan Laboratorium Biomedik .....................................72
Lampiran 7 : Surat Keterangan Kelaikan Etik ......................................................73
Lampiran 8 : Surat Keterangan Laboratorium Biomedik UB ...............................74
Lampiran 9 : Surat Keterangan Pembacaan PA ....................................................75
Lampiran 10 : Kartu Konsultasi Tugas Akhir .......................................................76
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DAFTAR PUSTAKA
Abdullah,et al, 2013, Expression of NF-kB and COX2 in Colorectal Cancer
among Native Indonesians: The Role of Inflammation in Colorectal
Carcinogenesis, ActaMedicaIndonesiana-The Indonesian Journal of
Internal Medicine, Halaman 187-192 (online)
http://www.inaactamedica.org/archives/2013/24045388.pdf diakses
25 Februari 2015.
Ahmad,et al, 2013, A review on therapeutic potential of Nigella sativa: A miracle
herb, Asian Pacific Journal of Tropical Biomedicine, Vol 3, No 5,
Halaman 337-352 (online) www.elsevier.com/locate/apjtb diakses 25
Februari 2015.
Baratawidjaya, Karnen G, Iris R, 2012, ImunologiDasaredisi 10,
FakultasKedokteranUniversitas Indonesia, Jakarta.
Biswell R., 2010, Kornea, In : Vaughan, Asbury, OftalmologiUmumEdisi 17,
EGC, Jakarta.
Clancy J, 2000, Basic Concepts in Immunology a Student's Survival Guide, the
McGraw-Hill companies, United states.
Crane et al, 2012, Mass Spectrometric Analyses of Phosphatidylcholines in Alkali-
Exposed Corneal Tissue, Investigative Ophthalmology & Visual
Science, Vol 53, No 11, Halaman 7122-7130 (online)
http://www.iovs.org/content/53/11/7122.full.pdf+html diakses 25
Februari 2015.
Dursumet al, 2012, Comparison of the effects of bevacizumab and ranibizumab
injection on corneal angiogenesis in an alkali burn induced model, Inj
J Ophthalmol, Vol , No 4, Halaman 448-451, (online)
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3428539/pdf/ijo-05-
04-448.pdf diakses2 Februari 2015.
53
Erdurmuset al, 2007, Inhibitory Effects of Topical Thymoquinone on Corneal
Neovascularization, Cornea, Vol 26, Halaman 715-719, (online)
http://www.researchgate.net diakses 28 Oktober 2014.
Eroschenko V P, 2012, Atlas HistologidiFiore, EGC, Jakarta.
Gagen D,.2011,Cellular and Molecular Mechanisms of Corneal Inflammation and
Wound Healing, Disertation Doctor of Philosophy, Houston College,
Houston.
Ibrahim, Zein S, et al, 2008, Protection by Nigella sativa against carbon
tetrachloride-induced downregulation of hepatic cytochrome P450
isozymes in rats, Japanese Journal of Veterinary Research, Vol 56, No
3, Halaman 119-128, (online)
http://eprints.lib.hokudai.ac.jp/dspace/bitstream/2115/35112/1/56-
3_p119-128.pdf diakses 28 Oktober 2014.
James, Bruce, et al, 2006, Leture notes: Oftalmologiedisikesembilan, Erlangga,
Jakarta.
Khan, et al, 2011, Anticancer Activities of Nigella Sativa (Black Cumin),Afr J
Tradit Complement Altern Med, Vol 8, Halaman 226-232, (online)
http://www.ajol.info/index.php/ajtcam/article/viewFile/67964/56060
diakses 24 Maret 2014.
Klausner, Eytan A, et al, 2007, Corneal gene therapy, Journal of Controlled
Release 124, Halaman 107–133, (online)
www.elsevier.com/locate/jconrel diakses 25 Februari 2015
Lin W W, Karin M, 2007. A Cytokine-Mediated Link Between Innate Immunity,
Inflamasi and Cancer, The Journal of Clinical Investigation, Vol 117,
No 5, Halaman 1175-1183 (online)
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1857251/pdf/JCI0731
537.pdf diakses 25 Februari 2015.
54
Morgan S R, 2014, Understanding the structural basis of corneal refractive
function and its modification via novel therapeutic approaches,
Thesis, Cardiff University, United Kingdom.
Notoarmodjo S, 2005, MetodologiPenelitianKesehatan, AsdiMahasatya, Jakarta.
O’Dea E, Alexande H, 2009, NF-kB Signaling, John Wiley & Sons Inc. Volume
1, Halaman 107-115, (online) www.wiley.com/wires/sysbio diakses
25 Desember 2014.
Paarakh P, 2010, Nigella sativa Linn – a Comprehensive Review, Indian Journal
of Natural Products and Resources, Vol 1, No 4, Halaman 409-429
(online)
http://probotanic.com/pdf_istrazivanja/crni_kumin/Sveobuhvatni%20
pregled%20zdravstvenih%20svojstava%20crnog%20kumina.pdf
diakses 24 Maret 2014.
Patil, et al, 2008, Heme Oxygenase-1 Induction Attenuates Corneal Inflammation
and Accelerates Wound Healing after Epithelial Injury. Investigative
Ophthalmology & Visual Science, Vol 49, No. 8, Halaman 3379-
3386, (online)
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2574648/pdf/nihms61
129.pdf diakses 25 Februari 2015
Prajapati, et al, 2010, Role of NFkB in Various Immunological & Inflammatory
Disorders, International Journal of Toxicological and
Pharmacological Research, Vol 2, No 1, Halaman 35-39, (online)
www.ijtpr.com diakses 25 Desember 2014.
Rajsekhar, et al, 2011, Pharmacognosy and Pharmacology of Nigella Sativa-A
Review, International Research Journal of Pharmacy, Vol 2, No 11,
Hal 36-39, (online) www.irjponline.com diakses 25 Desember 2014.
Riordan-Eva, Paul, 2010, AnatomidanEmbriologi Mata, In: Riordan-Eva P, John
P, Whitcher, Vaughan & Asbury OftalmologiUmum, EGC. Jakarta.
55
Salem M L, 2005, Immunomodulatory and therapeutic properties of the Nigella
sativa L. Seed, International Immunopharmacology, Vol 5. Halaman
1749–1770, (online) http://www.elsevier.com/locate/intimp
diakses 28 Oktober 2014.
Salim, et al, 2013, Thymoquinone Induces Mitochondria-Mediated Apoptosis in
Acute Lymphoblastic Leukaemia in Vitro, Molecules, Vol 18,
Halaman 11219-11240, (online) www.mdpi.com/journal/molecules
diakses 24 Maret 2014.
Salmani, et al, 2014, Aqueous Solubility and Degradation Kinetics of the
Phytochemical Anticancer Thymoquinone; Probing the Effects of
Solvents, pH and Light, Molecules, Vol 19, Halaman 5925-5939,
(online) www.mdpi.com/journal/molecules diakses 24 Maret 2014.
Sandhu KS, AC Rana, 2013, A Review of Plant Nigella Sativa: A Brief
Consideration of Its PharmacognosticCharacteers, Chemical
Constitutents and Therapeutic Benefits, Pharma Science Monitor An
International Journal Of Pharmaceutical Sciences, Vol 4, Issue 4, Hal
323-343, (online) www.pharmasm.com diakses 25 Desember 2014.
Sangi S, et al, 2014, Hepatoprotective Effects of Oleuropein. Thymoquinone and
Fruit of Phoenix Dactylifera on Ccl4, Induced Hepatotoxicity In Rats,
World Journal of Pharmaceutical ReseaRch, Volume 3, Issue 2,
Halaman 3475-3486, (online) www.wjpr.net diakses 25 Desember
2014.
Saryono, 2011,
MetodologiPenelitianKesehatanPenuntunPraktisBagiPemula.MitraCe
ndika Press, Yogyakarta.
Sethi G, et al, 2008, Targeting Nuclear Factor-KB Activation Pathway by
Thymoquinone: Role in Suppression of Antiapoptotic Gene Products
and Enhancement of Apoptosis, Mol Cancer Res, Vol 6, Halaman
1059-1070, (online)
56
http://mcr.aacrjournals.org/content/6/6/1059.full.pdf diakses 25
Februari 2015.
Siomek A, 2012, NF-κB Signaling Pathway and Free Radical Impact,
ActaBiochimicaPolonica, Vol 59, No 3, Halaman 323–331, (online)
www.actabp.pl diakses 25 Desember 2014.
Smolin, Throft’s, 2005, The Cornea Scientific Foundations and Clinical Practice
Fourth Edition,Lippincontt Williams & Wilkin, USA.
Sofia O, Rosy A, 2010, Effect of Genistein on Conjunctival Goblet Cells Density
and the Expression of Interleukin-1b in Ovariectomized Rats,
JurnalOftalmologi Indonesia, Vol 7, No. 3, Halaman 110-115,
(online)
http://journal.unair.ac.id/filerPDF/JOI%20Vol%207%20No%203%20
Juni%202010%20%28Ovi%20S%29.pdf diakses 28 Oktober 2014.
Stautd L, Micheal Karin, 2009, The Nuclear Factor NF-Kβ Pathway in
Inflammation, Cold Spring Harbor Laboratory Press, France.
Supranto J, 2007, StatistikTeoridanAplikasiEdisi 6, Erlangga, Jakarta.
Sutphin, et al, 2008.Basic and Clinical Science Course External Disease and
Cornea Section 8,American Academy of Opthalmologi, USA.
Tembhurne S V, et al, 2014, A Review on Therapeutic Potential of Nigella Sativa
(Kalonji) Seeds, Journal Of Medicinal Plants Research, Vol 8, No 3,
Hal 167-177, (online) http://www.academicjournals.org/JMPR
diakses 25 Desember 2014.
Toropainen E, 2007, Corneal Epithelial Cell Culture Model for Pharmaceutical
Studies, Doctoral Dissertation, University of Kuopio, Finland.
Yenita, 2010, PengaruhPemberianEkstrakJaheterhadap Kadar Malondialdehide
(MDA) Testis dan Diameter TubulusSeminiferusTestis Mencit yang
diberiPlumbumAsetat, JurnalKedokteranIbnuSina, Vol 5, No 3,
Halaman 106.
57
Wei Gu, et al, 2011, Effects of Amniotic Membrane Transplantation on Cytokines
Expression in Chemically Burned Rat Corneas, Int J Ophtthalmol,
Vol 4, No 1, Halaman 33-36, (online)
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3340686/pdf/ijo-04-
01-033.pdf diakses 25 Februari 2015.
Xiao, Gutian, Jing F, 2011, Nf-Kb and Cancer: A Paradigm Of Yin-Yang, Am J
Cancer Res, Vol 1, No 2, Halaman 192-221, (online) www.ajcr.us
diakses 25 Desember 2014.
Zhang,et al, 2005, Plasminogen Kringle 5 Inhibits Alkali-Burn–Induced Corneal
Neovascularization, Investigative Ophthalmology & Visual Science,
Vol 46, No 11, Halaman 4062-4071 (online)
http://www.ncbi.nlm.nih.gov/pubmed/16249481 diakses 25 Februari
2015.
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