الفهرس 
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Abstract
The Palms, Family Arecaceae Is One Of The Best Known Plant Families, Which Also Called Palmae, They Comprises 181 Genera With Around 2600 Species. Most Palms Are Distinguished By Their Large Evergreen Leaves, Known As Fronds, Arranged At The Top Of An Unbranched Stem. Palms Exhibit An Enormous Diversity In Physical characteristics And Inhabit Nearly Every Type Of Habitat Within Their Range, from Rainforests To Deserts (Chistenhusz And Byng, 2016). The Arecaceae Not Only Has Elegant And Predictable Shapes But Also Is Considered As A Commercial Source Of Nuts, Edible Fruits, Starchy Foods, Oils, Fibers And Woods. The Genus Livistona Is Widely Cultivated In Tropical Areas As A Landscaping Tree; It Is A Genus Of 36 Species Native To Australia, Horn Of Africa, And Southern And Southeastern Asia (Brown, 1810). There Are Four Livistona Species Growing In Egypt; Namely L. Australis, L. Chinensis, L. Decipiens And L. Rotundifolia And Three Species Are Growing In South China (Pei Et Al., 1991). L. Australis, L. Chinensis And L. Decipiens Were Chosen For This Study. They Have Been Traditionally Used For Their Analgesic, Hemostatic, Anti-Nasopharyngeal Carcinoma, Antichoriocarcinoma, Antiesophageal Cancer And Anti-Leukemic Properties (Zhao Et Al., 2001). Little Has Been Reported On L. Australis (Fam. Arecaceae). The Fruit Pulps Of L. Australis Exhibited Antioxidant, Antihyperlipidemic And Anti-Ulcer Activities (Kassem, Afifi Et Al. 2014). Previous Chemical Investigations On This Genus Reported Certain Flavonoids, Steroids, Amino Acids, Vitamins, Fatty Acids And Phenolics (Kaur And Singh, 2008; Li Et Al., 2008; Chen Et Al., 2007; Liu Et Al., 2009; Yaun Et Al., 2009; Mala And Dahot, 1994). This Study Aims To Isolate Phenolic Constituents Of L. Australis Fruits And Evaluate The Biological Activities Of L. Australis Leaves And Fruits.
Introduction And Literature Survey:
The Family Arecaceae (Palmae) Is A Large Botanical Family Comprises Several Genera Which Are Reported To Have Important Economic And Medicinal Uses. A Literature Survey Of Plants Of This Family Showed Presence Of Different Chemical Constituents With Interesting Biological Activities. So, L. Australis, L. Chinensis And L. Decipiens Family (Arecaceae) Were Chosen For Intensive Phytochemical And Biological Studies On Some Of These Plants.
Results And Discussion:
Part I: Genetic Profiling
DNA Fingerprinting Of Livistona Australis (R.Br.) Mart., Livistona Chinensis (Jacq.) R.Br. Ex Mart. And Livistona Decipiens Becc. Fresh Leaves: The Genomic DNA Of The Three Species Has Been Subjected To ISSR And Scot Analyses. Ten Decamer Olygonucleotide Primers Induced Successive Amplifications With A Large Number Of Polymorphic Bands. The Analysis Of The Amplicons Generated By ISSR And Scot Reactions Revealed That The Three Types Of L. Australis, L. Chinensis And L. Decipiens Species Were Reproducer And Gave Different Molecular Patterns. The Banding Patterns Produced By The Primers (HB-12) And (Scot 2) Generated The Highest Degree Of Polymorphism And Induced Unique Fragments. Therefore, These Two Primers Could Be Used To Discriminate Between Interspecific Species. However, Primers (HB-15), (Scot 6) And (Scot 8) Generated The Lowest Percent Of Polymorphism. These Three Primers Could Be Used For The Identification Of The Genotype. ISSR And Scot Techniques Are Widely Used To Fingerprint And Assess Genetic Diversity A Previously Reported By (Dahlberg, Zhang Et Al., 2002 And Salem Et Al., 2007).
Part II: Phytochemical Study
Chapter 1: Preliminary Phytochemical Screening Of Livistona Australis (R.Br.) Mart., Livistona Chinensis (Jacq.) R.Br. Ex Mart. And Livistona Decipiens Becc. Leaves And Fruits: Revealed That The Air Dried Leaves And Fruits Of The Three Species Contain Carbohydrates And/ Or Glycosides, Flavonoids, Sterols And/ Or Triterpenes from The Three Species While Coumarins, Saponins, Tannins And Volatile Oils Were Absent. Alkaloids And/Or Nitrogenous Bases Are Present In Leaves And Fruits Of L. Chinensis While L. Australis And L. Decipiens Didn’t Contain Any Traces.
Chapter2: Quantitative Estimation Of Total Phenolic And Total Flavonoids Content In Fruits And Leaves Of Livistona Australis (R.Br.) Mart., Livistona Chinensis (Jacq.) R.Br. Ex Mart. And Livistona Decipiens Becc.
Total Phenolics Content Was Determined In The Assigned Extracts By Folin–Ciocalteu Reagent, L. Australis Leaves Was Contained The Highest Phenolic Content (78.87 ± 0.313), Followed By The L. Decipiens Fruits (70.69 ± 1.732), Then L. Chinensis Fruits (44.50 ±0.9238), L. Australis Fruits (32.82 ± 0.7519) And L. Chinensis Leaves (19.72 ± 2.015) While L. Decipiens Leaves Contained The Least Phenolic Content (19.12 ± 0.6004).
Total Flavonoids Content Was Determined In The Assigned Extracts By Reaction With Sodium Nitrite, Followed By The Formation Of Colored Complex Of Flavonoid-Aluminum Using Aluminum Chloride That Was Spectrophotometrically Monitored At 510 Nm. L. Decipiens Fruits Contained The Highest Flavonoid Content (28.51± 0.29, Followed By The L. Chinensis Fruits (17.75 ± 1.039), Then L. Australis Fruits (12.81 ± 0.3551), L. Australis Leaves (8.16 ± 0.1906) And L. Decipiens Leaves (5.480 ± 0.05773) While L. Chinensis Leaves Contained The Least Flavonoid Content (4.82 ± 0.2214).
The Results Comparied Between The Fruits And Leaves Of Different Livistona Species, It Was Found That The Fruits Contain A Greater Amount Of Total Phenolic And Total Flavonoid Contents Than The Leaves Of The Different Livistona Species.
Chapter 3: HPLC- HR -ESI-MS/MS Profile Of Phenolic Compounds Of Leaves And Fruits Of Livistona Australis (R.Br.) Mart., Livistona Chinensis (Jacq.) R.Br. Ex Mart. And Livistona Decipiens Becc.: Metabolic Profiling Of The Secondary Metabolites Presents In L. Australis, L. Chinensis And L. Decipiens Leaves And Fruits Using LC–HRESIMS Was Conducted For Dereplication/Detection Of Putative Compounds Responsible For Various Activity Assigned To Livistona.Total Compounds In The Negative Ionization Mode ESI (-) Led To The characterization Of 65 Compounds, While Total Compounds In The Positive Ionization Mode ESI (+) Led To The characterization Of 42 Compounds. In The Negative Ionization Mode ESI (-), Leaves Contained 24 Compounds from L. Australis, 30 Compounds from L. Chinensis And 31 Compounds from L. Decipiens, While Fruits Contained 45 Compounds from L. Australis, 42 Compounds from L. Chinensis And 49 Compounds from L. Decipiens.
In The Positive Ionization Mode ESI (+), Leaves Contained 24 Compounds from L. Australis,19 Compounds from L. Chinensis And 22 Compounds from L. Decipiens, While Fruits Contained 26 Compounds from L. Australis, 27 Compounds from L. Chinensis And 33 Compounds from L. Decipiens Were Tentatively Identified. It Belonging To Different Classes Of Flavonoids And Phenolic Acids.
Chapter 4: Quantitative Estimation Of Nutritional Value Of Leaves And Fruits Of Livistona Australis (R.Br.) Mart.
Assessment Of Nutritional Values Revealed That Total Protein Content Was 6.74 And 5.13 G %, Crude Fiber Content Was 39.72 And 22.28 G %, And Total Lipid Content Was 2.57 And 2.93 G % For Leaves And Fruits Respectively. Five Minerals, Na, Ca, K, Fe And P Were Measured In Both Organs. Four Aflatoxins: B1, B2, G1 And G2 Were Also
Analyzed, And The Results Revealed That Leaves Were Free from Aflatoxins. The Nutritional Analysis Of Leaves And Fruits Showed That They Were Rich In Vitamin C, Fibers And Minerals (Sodium And Phosphorus For Leaves And Potassium, Calcium For Fruits) So They Can Be Used As Dietary Supplements.
Quantitative Estimation Of Amino Acids Of The Livistona Australis (R.Br.) Mart. Fruits.
The Results Revealed That Amino Acids Of L. Australis Fruits Showed That Fruits Were Rich In Cystine (10.21%) Contains The Largest Percentage Present In L. Australis Fruits Followed By Aspartic (0.51%), Glutamic (0.5%), Glycine (0.41%), Alanine (0.39 %), Valine (0.36%), Proline (0.32%), Lysine (0.31%), Argnine (0.28 %), Therionine (0.25 %), Phenylalanine (0.24%), Tyrosine (0.23%), Serine (0.21%), Isoleucine (0.2%), Hisitidine (0.13 %). Finally, Methionine (0.12%) Present In The Lowest Percentage.
Chapter 5: GC/MS Analysis Of Lipophilic Fractions Of Livistona Australis (R.Br.) Mart. Leaves And Fruits.
The Chemical Composition Of Lipophilic Fractions Of Leaves And Fruits Was Evaluated By Gas chromatography-Mass Spectrometry (GC/MS). The Results Of GC/MS Analysis Of Unsaponifiable Matter (USM) Of L. Australis Leaves Revealed The Presence Of 22 Compounds, The Major Compound Detected Was Phytol (16.44%) Followed By The Alkane Derivative Named 2-Phenyl Tridecane (11.41%). 1-Nonadecene And 1-Docosene Were The Major Unsaturated Hydrocarbons Detected With Area 5.40 And 5.24 %, Respectively. While The Results Of GC/MS Analysis Of Unsaponifiable Matter (USM) Of L. Australis Fruits Revealed Of 20 Compounds, The Major Compounds Detected Were Volatile Phenolic Constituents; Acetyl Eugenol (37.58%); Followed By Eugenol (29.61%). The Results Of GC/MS Analysis Of Fatty Acid Methyl Ester (FAME) Of L. Australis Leaves Revealed The Identification Of 26 Compounds, Hexadecanoic Acid Was The Major Methyl Ester Of Saturated Fatty Acid (25.86 %) Followed By 9-
Octadecadienoic Acid Methyl Ester (20.34 %). While The Results Of GC/MS Analysis Of Fatty Acid Methyl Ester (FAME) Of L. Australis Fruits Showed 44 Compounds, Octanoic Acid Was The Major Methyl Ester Of Saturated Fatty Acid (33.56 %) Followed By 7,10-Hexadecadienoic Acid Methyl Ester (28.06 %).
Chapter 6: Isolation, characterization And Structure Elucidation Of Flavonoids And Phenolic Acids from Livistona Australis (R.Br.) Mart. Fruits
1. Extraction And Fractionation
2. Investigation Of Ethyl Acetate And N-Hexane Fraction.
3. characterization And Identification Of Isolated Compounds.
Structure Elucidation Of The Pure Isolated Phenolic Compounds Were Performed Using Different Spectroscopic Techniques, UV, NMR And MS Spectrometry, And Their Structures Were Confirmed By Comparing Their Data With Those Previously Published In Literature. Six Known Compounds Were Isolated And Identified from The Air-Dried Pericarp Of L. Australis Fruits: One Fatty Acid from The Hexane Fraction And Five Flavonoids from The Ethyl Acetate Fraction. )These Compounds Have Been Isolated And characterized For The First Time from Livistona Australis (R.Br.) Mart. Fruits).
Table (54): Isolated Compounds from The Fruits Of Livistona Australis (R.Br.) Mart.
No. Compounds Structures
1 Quercetin 3-O-Α-D- Arabinopyranoside
2 Vanillic Acid
3 Protocatechuic Acid
4 Hyperoside
(Quercetin3-O-Β-D-Galactopyranoside)
5 Gallic Acid
6 Dodecanoic Acid
Part III: In Vitro Biological Study
Chapter1: 1.1. Evaluation Of Certain Bioactivities Of Livistona Australis (R.Br.) Mart. Leaves And Fruits.
1.1.1. Anti-Ageing Activity: The Results Revealed That Leaves And Fruits Extract Of L. Australis Was The Most Potent Tolemarse Activity As Compared The Reference Drug Curcumin.
1.1.2. Anti-Cholinesterase Activity: The Results Revealed That Leaves Extract Of L. Australis Was Potent As Anti-Acetylcholinesterase Compared To The Standard, Medicinal Plants Rich With Flavonoids Are Promising Class Of Natural Compound In Management Of Alzheimer Disease (AD).
1.1.3. Anti-Diabetic Activity: The Results Showed That L. Australis Leaves Extract Was The Most Significant Action Withic50 9.08±0.448 G/Ml As Compared The Reference Drug Sitagliptin.
1.1.4. Anti-Inflammatory Activity: The Results Revealed That The Leaves Extract Of L. Australis Is The Most Potent Anti-Inflammatory Activity As Compared The Reference Drug Ibuprofen.
1.1.5. Anti-Microbial Activity: L. Australis Leaves No Activity Against Aspergillus Fumigatus While L. Australis Fruits Was Very Weak Effect Against Aspergillus Fumigatus And Candida Albicans. On The Other Hand, The Antibacterial Test Revealed That L. Australis Leaves And Fruits Were Very Weak Effect Against Gram Positive Bacteria (Staphylococcus Aureus And Bacillus Subtilis) And Gram Negatvie Bacteria (Proteus Vulgaris And Escherichia Coli.).
Chapter 2: Evaluation Of Anticaner Activity Of Livistona Australis (R.Br.) Mart. Fruits And Leaves
The Fruits And Leaves Extract Of L. Australis Exhibit More Anti Cancer Activity Against Cervical Cancer Cells (Hela Cells) Than Fibrosarcoma Cells (HT-1080 Cells) And Human Colorectal Adenocarcinoma Cell Line (HT-29 Cells).
The Extract Of L. Australis Fruits And Leaves Exhibit No Significant Anticancer Activity Against Lung Carcinoma Cells (A-549 Cells) And Human Muscle Carcinoma (Rdcells).
The Extract Of L. Australis Leaves Showed More Anticancer Activity Against Cervical Cancer Cells (Hela Cells), Fibrosarcoma Cells (HT-1080 Cells) And Human Colorectal Adenocarcinoma Cell Line (HT-29 Cells) Than L. Australis Fruits.
Our Findings Of These Biological Activities Of Leaves And Fruits Extract Of Livistona Australis (R.Br.) Mart. And Livistona Decipiens Becc. Are Reported Here For The First Time.
Chapter 3: Molecular Docking And Evaluation Of Secondary Metabolites Of Livistona Decipiens Leaves And Fruits As Potential Inhibitors Of SARS-Cov-2
1- Molecular Docking Study
The Flexible Docking Results Showed That The Three Flavonoid Glucosides Isoquercetin (-8.2), Vitexin (-7.6), And Isoorientin (-7.6) Were The Top Ranked Ligands Followed By The Five Flavonoid Aglycones Such As Tricin (-6.7), Quercetin (-6.4), Luteolin (-6.2), Epiafzelechin (-6.2) And Catechin (-5.8). The High Binding Affinity Of These Three Flavonoid Glucosides To COVID-19 Virus (Mpro) Can Be Explained By The Ability Of Their Saccharide Moiety To Make Strong Interactions With Some Key Amino Acids.
2-Evaluation Of Secondary Metabolites Of Livistona Decipiens Leaves And Fruits As Potential Inhibitors Of SARS-Cov-2
SARS-Cov-2 Is The Main Causative Agent Of COVID-19 Pneumonia Pandemic Attack That Emerged In Wuhan. Discovering Known Natural Agent That Can Inhibit SARS-Cov-2 Main Protease (Mpro) Will Lead To Controlling Viral Transcription And Replication. Twelve Known Compounds Were Identified from The Alcoholic Extract Of L. Decipiens Leaves And Fruits Using LC–HRESIMS. The Individual Components Were Belonging To Several Classes Including Six Flavonoids (Five Aglycones And Three Glucosides), Phenolic Acids (Two Caffeic Acid And One Benzoic Acid Derivatives) And A Methoxylated Phenol. Identification Of The Annotated Compounds Enabled Testing The Relationship Between Their Interactions With Receptor In The N3 Binding Site In Mpro And Their Structure. In Silico Structure-Based Screening And ADME Study Revealed That Tricin Is A Potential Anti-COVID Lead Compound. The Alcoholic Extract Was Shown To
Inhibit SARS-Cov-2 Through In Vitro Culture And RT-PCR Test With EC50=0.122 And 1.53 Μg/Ml For Leaves And Fruits Extracts, Respectively When Compared With That Of Remdesivir (0.002 Μg/Ml). This Study Provides Scientific Basis For Demonstrating Health Benefits Of L. Decipiens Use. In Vivo Testing Is Required To Verify The Observed Effect Of The Active Phytochemicals.
3CL-Protease Inhibition Assay Was Used To Assess The Effect Of L. Decipens Leaves And Fruits Extracts On Inhibition Of The Main Protease (Mpro) Enzyme Which Is Responsible For Viral Replication And Transcription And Consequently Affects Human SARS-Cov-2. Percentage Of Inhibition Of SARS-Cov-2 Replication Was Determined Through Estimation Of Viral RNA Concentration Using Quantitative RT-PCR (Real Time-Polymerase Chain Reaction) Assay Remdesivir Was Used As A Reference Standard. The 3CL Protease Inhibition Assay Was Performed As A Preliminary Practical Evaluation. L. Decipiens Exhibited A Significant Inhibitory Action Against 3CL-Protease With IC50=15.58±0.79 And 41.81±2.12 Μg/Ml For Leaves And Fruits Extracts. This Result Is Consistent With The Stabilized Binding Of Compounds (1 - 12) At The Binding Site Of Mpro.