Zeytin Yaprağı Ekstraktı ve Oleuropeinin Antioksidan, Antimikrobiyal Aktivitesi, Gıdalarda Kullanım Olanakları

Zeytin ağacı yetiştiriciliği ve zeytin işleme endüstrisinin yan ürünlerinden biri olan zeytin yaprağı yüzyıllar boyunca geleneksel halk hekimliğinde kullanılmıştır. Son yıllarda yapılan çalışmalarda ise zeytin yaprağının birçok biyoaktiviteye sahip olduğu belirlenmiş ve bu etkiler içeriğinde bulunan yüksek fenolik bileşik miktarı ile ilişkilendirilmiştir. Zeytin yaprağında bulunan en baskın fenolik bileşik elenolik asit ve 3,4-dihidroksifeniletanolün heterozidik esteri olan oleuropeindir. Bundan dolayı, zeytin yaprağından katma değeri yüksek bileşiklerin ekstrakte edilmesine yönelik bazı çalışmalar gerçekleştirilmiş ve elde edilen ham fenolik ekstrakt ve oleuropeinin antioksidan ve antimikrobiyal aktiviteleri incelenmiştir. Ayrıca sentetik gıda koruyucularının yan etkisi ve toksisitesine karşı şüphelerin artmasından dolayı zeytin yaprağı ekstraktı ve oleuropeinin gıda sanayiinde kullanım olanaklarının arttırılmasına yönelik bazı çalışmalarda gerçekleştirilmiştir. Bu derlemede, zeytin yaprağı fenolik bileşikleri ve zeytin yaprağından fenolik bileşik ekstraksiyonu, zeytin yaprağı ekstraktı ve oleuropeinin antioksidan kapasitesi, antimikrobiyal aktivitesi ile gıdalarda kullanım olanaklarının incelenmesi amaçlanmıştır.

Antioxidant, Antimicrobial Activity of Olive Leaf Extract and Oleuropein, Their Possibilities Usage in Foods

Olive leaves which is one of the by-products of olive tree cultivation and olive processing industry,have been used in traditional folk medicine for centuries. In recent studies, it has been determinedthat olive leaf has many bioactivities and these effects have been associated with high phenoliccompound content. The most dominant phenolic compound of olive leaf is oleuropein, which is theheterosidic ester of elenolic acid and 3,4-dihydroxyphenylethanol. Therefore, some studies havebeen carried out for extracting high value added compounds from olive leaves in recent years.Antioxidant and antimicrobial activities of crude phenolic extract and oleuropein have beeninvestigated. Moreover, some studies have been carried out to increase the possibility of using oliveleaf extract and oleuropein in food industry due to increasing suspicion of side effects and toxicityof synthetic food preservatives. In this review, it was aimed to investigate phenolic compounds of olive leaf extract, phenolic compound extraction from olive leaf as well as antioxidant, antimicrobial activity of olive leaf extract and oleuropein and possibilities of use in foods.

PDF

___

Abaza L, Youssef NB, Manai H, Haddada FM, Methenni K, Zarrouk M. 2011. Chétoui olive leaf extracts: influence of the solvent type on phenolics and antioxidant activities. Grasas y Aceites, 62(1): 96-104.
Ahmad-Qasem MH, Cánovas J, Barrajón-Catalán E, Micol V, Andrés J, García-Pérez JV. 2013. Kinetic and compositional study of phenolic extraction from olive leaves (var. Serrana) by using power ultrasound. Innovative Food Science and Emerging Technologies, 17: 120-129.
Alagna F, Mariotti R, Panara F, Caporali S, Urbani S, Veneziani G, Esposto S, Taticchi A, Rosati A, Rao R, Perrotta G, Servili M, Baldoni L. 2012. Olive phenolic compounds: metabolic and transcriptional profiling during fruit development. BMC Plant Biology, 12(1): 162.
Al-Azzawie HF, Alhamdani MSS. 2006. Hypoglycemic and antioxidant effect of oleuropein in alloxan-diabetic rabbits. Life sciences, 78(12): 1371-1377.
Altıok E, Bayçın D, Bayraktar O, Ülkü S. 2008. Isolation of polyphenols from the extracts of olive leaves (Olea europaea L.) by adsorption on silk fibroin. Separation and Purification Technology, 62(2): 342-348.
Andreadou I, Sigala F, Iliodromitis EK, Papaefthimiou M, Sigalas C, Aligiannis N, Savvari P, Gorgoulis V, Papalabros E, Kremastinos DT. 2007. Acute doxorubicin cardiotoxicity is successfully treated with the phytochemical oleuropein through suppression of oxidative and nitrosative stress. Journal of Molecular and Cellular Cardiology, 42(3): 549-558.
Anonim 2017a. http://www.fao.org/faostat/en/#data/QC. [Erişim Tarihi: 10/07/2019].Anonim, 2017b. https://pubchem.ncbi.nlm.nih.gov/compound/ Oleuropein. [Erişim Tarihi: 5/07/2019].
Aouidi F, Okba A, Hamdi M. 2017. Valorization of functional properties of extract and powder of olive leaves in raw and cooked minced beef meat. Journal of the Science of Food and Agriculture, 97(10): 3195-3203.
Aytul KK. 2010. Antimicrobial and antioxidant activities of olive leaf extract and its food applications (Doktora Tezi), İzmir Yüksek Teknoloji Enstitüsü, İzmir-Türkiye.
Azmir J, Zaidul ISM, Rahman MM, Sharif KM, Mohamed A, Sahena F, Jahurul MHA, Ghafoor K, Norulaini NAN, Omar AKM. 2013. Techniques for extraction of bioactive compounds from plant materials: a review. Journal of Food Engineering, 117(4): 426-436.
Basmacıoğlu-Malayoğlu HB, Aktaş B. 2011. Zeytinyağı işleme yan ürünlerinden zeytin yaprağı ile zeytin karasuyunun antimikrobiyal ve antioksidan etkileri. Hayvansal Üretim, 52(1): 49-58.
Başoğlu F. 2010. Yemeklik Yağ Teknolojileri. Dora Yayın Dağıtım, Bursa. Benavente-Garcia O, Castillo J, Lorente J, Ortuno A, Del Rio JA. 2000. Antioxidant activity of phenolics extracted from Olea europaea L. Leaves. Food Chemistry, 68(4): 457-462.
Bıçakçı A, Altunoğlu MK, Tosunoğlu A, Çelenk S, Canitez Y, Malyer H, Sapan N. 2009. Türkiye’de Oleaceae familyasına ait allerjenik Olea (zeytin ağacı) ve Fraxinus (dişbudak ağacı) polenlerinin havadaki dağılımları. Asthma Allergy Immunol, 7: 133-146.
Bisignano G, Tomaino A, Cascio RL, Crisafi G, Uccella L, Saija A. 1999. On the In-vitro Antimicrobial Activity of Oleuropein and Hydroxytyrosol. Journal of Pharmacy and Pharmacology, (51): 971-974.
Boskou D. 1996. Olive oil chemistry and technology. History and characteristics of the olive tree. AOCS Press, Champaign, İllinois, 1-6.
Boss A, Bishop K, Marlow G, Barnett M, Ferguson L. 2016. Evidence to support the anti-cancer effect of olive leaf extract and future directions. Nutrients, 8(8): 513.
Bouaziz M, Sayadi S. 2005. Isolation and evaluation of antioxidants from leaves of a Tunisian cultivar olive tree. European Journal of Lipid Science and Technology, 107(7‐ 8): 497-504.
Bouaziz M, Feki I, Ayadi M, Jemai H, Sayadi S. 2010. Stability of refined olive oil and olive‐pomace oil added by phenolic compounds from olive leaves. European Journal of Lipid Science and Technology, 112(8): 894-905.
Bulotta S, Corradino R, Celano M, D’Agostino M, Maiuolo J, Oliverio M, Procopio A, Iannone M, Rotiroti D, Russo D. 2011. Antiproliferative and antioxidant effects on breast cancer cells of oleuropein and its semisynthetic peracetylated derivatives. Food Chemistry, 127(4): 1609-1614.
Bulotta S, Celano M, Lepore SM, Montalcini T, Pujia A, Russo D. 2014. Beneficial effects of the olive oil phenolic components oleuropein and hydroxytyrosol: focus on protection against cardiovascular and metabolic diseases. Journal of Translational Medicine, 12(1): 219.
Cicerale S, Lucas LJ, Keast RSJ. 2012. Antimicrobial, antioxidant and antiinflammatory phenolic activities in extra virgin olive oil. Current Opinion in Biotechnology, 23(2): 129-135.
Cifa D, Skrt M, Pittia P, Di Mattia C, Poklar Ulrih N. 2018. Enhanced yield of oleuropein from olive leaves using ultrasound‐assisted extraction. Food Science & Nutrition, 6:1128-1137.
Czerwińska M, Kiss AK, Naruszewicz M. 2012. A comparison of antioxidant activities of oleuropein and its dialdehydic derivative from olive oil, oleacein. Food Chemistry, 131(3):940-947.
De Leonardis A, Aretini A, Alfano G, Macciola V, Ranalli G. 2008. Isolation of a hydroxytyrosol-rich extract from olive leaves (Olea europaea L.) and evaluation of its antioxidant properties and bioactivity. European Food Research and Technology, 226(4): 653-659.
Difonzo G, Russo A, Trani A, Paradiso VM, Ranieri M, Pasqualone A, Summo C, Tamma G, Silletti R, Caponio F. 2017. Green extracts from Coratina olive cultivar leaves: Antioxidant characterization and biological activity. Journal of Functional Foods, 31: 63-70.
Dominciano LCC, Oliveira CAF, Lee SH, Corassin CH. 2016. Individual and combined antimicrobial activity of oleuropein and chemical sanitizers. Journal of Food Chemistry Nanotechnology, 2(3): 124-127.
Domitrović R, Jakovac H, Marchesi VV, Šain I, Romić Ž, Rahelić D. 2012. Preventive and therapeutic effects of oleuropein against carbon tetrachloride induced liver damage in mice. Pharmacological Research, 65(4): 451-464.
Dua S, Bhat ZF, Kumar S. 2015. Effect of oleuropein on the oxidative stability and storage quality of Tabaq-Maz, fried mutton ribs. Food Bioscience, (12): 84-92.
Elgin-Cebe G, Konyalıoğlu S, Zeybek U. 2012. Olea europaea var. Europaea (Zeytin) Yaprak İnfüzyonunun Antioksidan Etkisi. Ege Üniversitesi Ziraat Fakültesi Dergisi, 49(3): 209-212.
Erbay Z, Icier F. 2010. The importance and potential uses of olive leaves. Food Reviews International, 26(4): 319-334.
Farag RS, Mahmoud EA, Basuny AM. 2007. Use crude olive leaf juice as a natural antioxidant for the stability of sunflower oil during heating. International Journal of Food Science & Technology, 42(1): 107-115.
Furneri PM, Marino A, Saija A, Uccella N, Bisignano G. 2002. In vitro antimycoplasmal activity of oleuropein. International Journal of Antimicrobial Agents, (20): 293-296.
Goulas V, Exarchou V, Troganis AN, Psomiadou E, Fotsis T, Briasoulis E, Gerothanassis IP. 2009. Phytochemicals in olive-leaf extracts and their antiproliferative activity against cancer and endothelial cells. Molecular Nutrition & Food Research, (53): 600-608.
Haddadin MSY. 2010. Effect of olive leaf extracts on the growth and metabolism of two probiotic bacteria of intestinal origin. Pakistan Journal of Nutrition, 9(8): 787-793.
Hadrich F, Mahmoudi A, Bouallagui Z, Feki I, Isoda H, Feve B, Sayadi S. 2016. Evaluation of hypocholesterolemic effect of oleuropein in cholesterolfed rats. Chemico-Biological Interactions, (252): 54-60.
Hassen I, Casabianca H, Hosni K. 2015. Biological activities of the natural antioxidant oleuropein: Exceeding the expectation-A mini-review. Journal of Functional Foods, 18: 926-940.
Hayes JE, Stepanyan V, Allen P, O’grady MN, Kerry JP. 2010. Effect of lutein, sesamol, ellagic acid and olive leaf extract on the quality and shelf-life stability of packaged raw minced beef patties. Meat Science, 84(4): 613-620.
Herrero M, Temirzoda TN, Segura-Carretero A, Quirantes R,Plaza M, Ibañez E. 2011. New possibilities for the valorization of olive oil by-products. Journal of Chromatography A, 1218(42): 7511-7520.
Hugo WB, Bloomfield SF. 1971. Studies on the mode of action of the phenolic antibacterial agent fentichlor against Staphylococcus aureus and Escherichia coli II. The effects of fentichlor on the bacterial membrane and the cytoplasmic constituents of the cell. Journal of Applied Bacteriology, 34(3): 579-591.
Iraqı R, Vermeulen C, Benzekrı A, Bouseta A, Collin S. 2005. Screening for Key Odorants in Moroccan Green Olives by Gas Chromatography-Olfactometry/Aroma Extract Dilution Analysis. Journal of Agricultural Food Chemistry, (53): 1179-1184.
Jaber H, Ayadi M, Makni J, Rigane G, Sayadi S, Bouaziz M.2012. Stabilization of refined olive oil by enrichment with chlorophyll pigments extracted from Chemlali olive leaves. European Journal of Lipid Science and Technology, 114(11):1274-1283.
Japón-Luján R, Luque-Rodríguez JM, De Castro ML. 2006a. Multivariate optimisation of the microwave-assisted extraction of oleuropein and related biophenols from olive leaves. Analytical and Bioanalytical Chemistry, 385(4): 753-759.
Japón-Luján R, Luque-Rodríguez JM, De Castro ML. 2006b. Dynamic ultrasound-assisted extraction of oleuropein and related biophenols from olive leaves. Journal of Chromatography A, 1108(1): 76-82.
Jemai H, Bouaziz M, Fki I, El Feki A, Sayadi S. 2008. Hypolipidimic and antioxidant activities of oleuropein and its hydrolysis derivative-rich extracts from Chemlali olive leaves. Chemico-Biological Interactions, 176(2-3): 88-98.
Jemai H, El Feki A, Sayadi S. 2009. Antidiabetic and antioxidant effects of hydroxytyrosol and oleuropein from olive leaves in alloxan-diabetic rats. Journal of Agricultural and Food Chemistry, 57(19): 8798-8804.
Jiménez P, García P, Bustamante A, Barriga A, Robert P. 2017. Thermal stability of oils added with avocado (Persea americana cv. Hass) or olive (Olea europaea cv. Arbequina) leaf extracts during the French potatoes frying. Food Chemistry, 221: 123-129.
Korukluoglu M, Sahan Y, Yigit A, Karakas R. 2006. Antifungal activity of olive leaf (Olea Europaea L.) extracts from the Trilye region of Turkey. Annals of Microbiology, 56(4): 359.
Korukluoğlu M, Şahan Y, Yiğit A, Tümay-Özer E, Gücer Ş. 2010. Antibacterial activity and chemical constitutions of Olea europaea L. leaf extracts. Journal of Food Processing and Preservation, (34): 383-396.
Kotyzová D, Hodková A, Eybl V. 2011. The effect of olive oil phenolics-Hydroxytyrosol and oleuropein on antioxidant defence status in acute arsenic exposed rats. Toxicology Letters, (205): S222.
Kurt Ş, Ceylan HG. 2017. Effects of olive leaf extract on the oxidation stability and microbiological quality of dry fermented sausage (sucuk). Carpathian Journal of Food Science & Technology: 9(4): 178-188.
Liu Y, McKeever LC, Malik NS. 2017. Assessment of the antimicrobial activity of olive leaf extract against foodborne bacterial pathogens. Frontiers in Microbiology, 8: 113.
Malheiro R, Casal S, Teixeira H, Bento A, Pereira JA. 2013. Effect of olive leaves addition during the extraction process of overmature fruits on olive oil quality. Food and Bioprocess Technology, 6(2): 509-521.
Marhamatizadeh MH, Ehsandoost E, Gholami P, Mohaghegh MD. 2013. Effect of olive leaf extract on growth and viability of Lactobacillus acidophilus and Bifidobacterium bifidum for production of probiotic milk and yoghurt. International Journal of Farming Allied Sciences, 2: 572-578.
Markin D, Duek L, Berdicevsky I. 2003. In vitro antimicrobial activity of olive leaves. Antimikrobielle Wirksamkeit von Olivenblättern in vitro. Mycoses, 46(3‐4): 132-136.
Mohamed MB, Guasmi F, Ali SB, Radhouani F, Faghim J, Triki T, Kammoun NG, Baffi C, Lucini L, Benincasa C. 2018. The LC-MS/MS characterization of phenolic compounds in leaves allows classifying olive cultivars grown in South Tunisia. Biochemical Systematics and Ecology, 78: 84-90.
Moudache M, Nerín C, Colón M, Zaidi F. 2017. Antioxidant effect of an innovative active plastic film containing olive leaves extract on fresh pork meat and its evaluation by Raman spectroscopy. Food Chemistry, 229: 98-103.
Omar SH. 2010a. Cardioprotective and neuroprotective roles of oleuropein in olive. Saudi Pharmaceutical Journal, (18): 111-121.
Omar SH. 2010b. Oleuropein in Olive and its Pharmacological Effects. Scientia Pharmaceutica, (78): 133-154.
Oskay D, Oskay M. 2009. Bitki sekonder metabolitlerinin biyoteknolojik önemi. Ecological Life Sciences, 4(2): 31-41.
Özcan MM, Matthäus B. 2017. A review: benefit and bioactive properties of olive (Olea europaea L.) leaves. European Food Research and Technology, 243(1): 89-99.
Peker H, Arslan S. 2017. Effect of Olive Leaf Extract on the Quality of Low Fat Apricot Yogurt. Journal of Food Processing and Preservation, 41(5): e13107.
Pereira AP, Ferreira IC, Marcelino F, Valentão P, Andrade PB, Seabra R, Estevinho L, Bento A, Pereira JA. 2007. Phenolic compounds and antimicrobial activity of olive (Olea europaea L. Cv. Cobrançosa) leaves. Molecules, 12(5): 1153-1162.
Putnik P, Barba FJ, Španić I, Zorić Z, Dragović-Uzelac V, Kovačević DB. 2017. Green extraction approach for the recovery of polyphenols from Croatian olive leaves (Olea europea). Food and Bioproducts Processing, 106: 19-28.
Qabaha K, AL-Rimawi F, Qasem A, Naser SA. 2018. Oleuropein is responsible for the major anti-inflammatory effects of olive leaf extract. Journal of Medicinal Food, 21(3): 302-305.
Qadir NM, Ali KA, Qader SW. 2016. Antidiabetic Effect of Oleuropein from Olea europaea Leaf against Alloxan Induced Type 1 Diabetic in Rats. Brazilian Archives of Biology and Technology, 59.
Romani A, Mulinacci N, Pinelli P, Vincieri FF, Cimato A. 1999. Polyphenolic content in five Tuscany cultivars of Olea europaea L. Journal of Agricultural and Food Chemistry, 47 (3): 964-967.
Romero M, Toral M, Gómez-Guzmán M, Jiménez R, Galindo P, Sánchez M, Olivares M, Galvez J, Duarte J. 2016. Antihypertensive effects of oleuropein-enriched olive leaf extract in spontaneously hypertensive rats. Food & Function, 7(1): 584-593.
Ruiz-Barba JL, Garrido‐Fernandez A, Jimenez‐Diaz R. 1991. Bactericidal action of oleuropein extracted from green olives against Lactobacillus plantarum. Letters in Applied Microbiology, 12(2): 65-68.
Ryan D, Antolovich M, Prenzler P, Robards K, Lavee S. 2002. Biotransformations of phenolic compounds in Olea europaea L. Scientia Horticulturae, 92(2): 147-176.
Sabry OM. 2014. Beneficial health effects of olive leaves extracts. Journal of Natural Sciences Research, 4(19): 1-9.
Salah MB, Abdelmelek H, Abderraba M. 2012. Study of phenolic composition and biological activities assessment of olive leaves from different varieties grown in Tunisia. Medicinal Chemistry, 2(5): 107-111.
Sanchez JC, Alsina MA, Herrlein MK, Mestres C. 2007. Interaction between the antibacterial compound, oleuropein, and model membranes. Colloid Polymer Science, (285): 1351-1360.
Souilem S, Fki I, Kobayashi I, Khalid N, Neves MA, Isoda H, Sayadi S, Nakajima M. 2017. Emerging technologies for recovery of value-added components from olive leaves and their applications in food/feed industries. Food and Bioprocess Technology, 10(2): 229-248.
Sudjana AN, D’Orazio C, Ryan V, Rasool N, Ng J, Islam N, Riley TV, Hammer KA. 2009. Antimicrobial activity of commercial Olea europaea (olive) leaf extract. International Journal of Antimicrobial Agents, 33(5): 461-463.
Şahin S. 2011. Zeytin Ağacı Yapraklarından Süperkritik-CO2 ile Ekstrakt Eldesi ve Bileşimindeki Oleuropein Miktarının İncelenmesi. (Doktora Tezi), İstanbul Üniversitesi, Fen Bilimleri Enstitüsü, İstanbul.
Şahin S, Bilgin M, Dramur MU. 2011. Investigation of oleuropein content in olive leaf extract obtained by supercritical fluid extraction and soxhlet methods. Separation Science and Technology, 46(11): 1829-1837.
Şahin S, Bilgin M, Sayım E, Güvenilir B. 2017. Effects of natural antioxidants in the improvement of corn oil quality: olive leaf vs. lemon balm. International Journal of Food Science &Technology, 52(2): 374-380.
Savournin C, Baghdikian B, Elias R, Dargouth-Kesraoui F, Boukef K, Balansard, G. 2001. Rapid high-performance liquid chromatography analysis for the quantitative determination of oleuropein in Olea europaea leaves. Journal of Agricultural and Food Chemistry, 49(2): 618-621.
Soler-Rivas C, Espín JC, Wichers HJ. 2000. Oleuropein and related compounds. Journal of the Science of Food and Agriculture, 80(7): 1013-1023.
Taamalli A, Arráez-Roman D, Barrajón-Catalán E, Ruiz-Torres V, Pérez-Sánchez A, Herrero M, Ibanez E, Micol V, Zarrouk M, Segura-Carretero A, Fernandez-Gutierrez A. 2012. Use of advanced techniques for the extraction of phenolic compounds from Tunisian olive leaves: phenolic composition and cytotoxicity against human breast cancer cells. Food and Chemical Toxicology, 50(6): 1817–1825.
Talhaoui N, Taamalli A, Gómez-Caravaca AM, FernándezGutiérrez A, Segura-Carretero A. 2015. Phenolic compounds in olive leaves: Analytical determination, biotic and abiotic influence, and health benefits. Food Research International,77: 92-108.
Vogel P, Kasper Machado I, Garavaglia J, Zani VT, de Souza D, Morelo SDB. 2014. Polyphenols benefits of olive leaf (Olea europaea L) to human health. Nutricion Hospitalaria, 31(3): 1427-1433.
Wainstein J, Ganz T, Boaz M, Bar Dayan Y, Dolev E, Kerem Z, Madar Z. 2012. Olive leaf extract as a hypoglycemic agent in both human diabetic subjects and in rats. Journal of Medicinal Food, 15(7): 605-610.
Winkelhausen E, Pospiech R, Laufenberg G. 2005. Antifungal activity of phenolic compounds extracted from dried olive pomace. Bulletin of the Chemists and Technologists of Macedonia, 24(1): 41-46.
Xu Y, Burton S, Kim C, Sismour E. 2016. Phenolic compounds, antioxidant, and antibacterial properties of pomace extracts from four Virginia‐grown grape varieties. Food Science & Nutrition, 4(1): 125-133.
Xynos N, Papaefstathiou G, Psychis M, Argyropoulou A, Aligiannis N, Skaltsounis AL. 2012. Development of a green extraction procedure with super/subcritical fluids to produce extracts enriched in oleuropein from olive leaves. The Journal of Supercritical Fluids, (67): 89-93.
Yıldız G, Uylaşer V. 2011. Doğal bir antimikrobiyel: oleuropein. Uludağ Üniversitesi Ziraat Fakültesi Dergisi, 25(1): 131-142.
Zehra A, Naqvi SBS, Ali SQ. 2016. In Vitro Evaluation of Antimicrobial Effect of Extracts of Medicinal Plant’s Leaves. Journal of Medical Microbiology Diagnosis, 5: 236.
Zorić N, Kopjar N, Kraljić K, Oršolić N, Tomić S, Kosalec I. 2016. Olive leaf extract activity against Candida albicans and C. dubliniensis–the in vitro viability study. Acta Pharmaceutica, 66(3): 411-421.
Zribi A, Gargouri B, Jabeur H, Rebaï A, Abdelhedi R, Bouaziz M. 2013. Enrichment of pan‐frying refined oils with olive leaf phenolic‐rich extract to extend the usage life. European Journal of Lipid Science and Technology, 115(12): 1443-1453.