Thesis (PhD)--Stellenbosch University, 2014. / ENGLISH ABSTRACT: Salvia africana lutea is one of 26 Sage species indigenous to Southern Africa from a total of 900 worldwide. The genus Salvia belongs to the Lamiaceae family. Labeled a ‘broad spectrum remedy’ S. africana lutea amongst other sage species is medicinally important. Reports are many highlighting its benefits, which include from alleviating coughs and colds to gynaecological complaints. Studies have revealed in vitro antimicrobial, anti-cancer and antioxidant activity. Plant secondary metabolites fundamentally have a strong bearing on the phytochemical activities a plant may possess. Consequently the environment indirectly affects the phytochemical properties as it influences the variation in the plant metabolome via plant-environment interactions. Five S. africana lutea plant populations, within the Western Cape province of South Africa were sampled and chemotypes and bioactivity tested. Four populations were wild growing in protected areas namely; Brackenfell, Koeberg, Silwerstroomstrand and Yzerfontein, while the fifth was a garden growing population from Stellenbosch. Using gas chromatography hyphenated with mass spectrometry (GC-MS), compounds such as monosaccharides, carboxylic acids and fatty acids were detected. Variation of compounds identified with 80% certainty was compared across all populations. Stellenbosch population showed some compounds that were not present in the other four sites. These compounds were namely; propanoic acid, rythronic acid, 2-keto-1-gluconic acid and 1,3-dibromobicyclon, while this population also did not have xylitol that was detected in all the other four populations. To consolidate the GC-MS findings, analysis on the metabolite profiles (utilizing liquid chromatography linked with mass spectrometry (LC-MS) and nuclear magnetic resonance (1H NMR)) was done. Principal component analysis (PCA) was applied to the NMR data. The partial least squaresdiscriminant analysis (PLS-DA) was used to integrate LC-MS and NMR data sets. All statistics were performed with the SIMCA-P+ 12.0 software. By integrating LC-MS and 1H NMR analyses, large chemotype differences leading to samples grouping by site, suggested strong plant-environment interactions as factors influencing metabolite composition. Signals distinguishing the Stellenbosch profile were in the aromatic part of the 1H NMR spectra. Antimicrobial activity was tested against two Fusarium species. Fusarium is a plant pathogenic species that causes large agricultural losses particularly in the maize crop, one of the staple foods in the African continent. Some species also produce mycotoxins in infected crop and lead to a significant increase in the risk factor of cancers when contaminated foods are consumed. Four high-mycotoxin producing strains from two species F. verticillioides (MRC 826 and MRC 8267); F. proliferatum (MRC 7140 and MRC 6809) were utilized in all in vitro antifungal assays in this study. A preliminary assay using dichloromethane: methanol (1:1 v/v) crude plant extracts of the five populations; Stellenbosch, Brackenfell, Koeberg, Silwerstoomstrand and Yzerfontein, from 2009 and 2011. All test samples exhibited good activity as the minimum inhibitory concentrations (MIC) values ranged from 0.031 mg ml-1 to 0.5 mg ml-1, values below the latter are regarded as strong inhibitors. The Stellenbosch extracts were the most active for both 2009 and 2011 collections, with the best activity against F. verticillioides MRC 8267 and MRC 826 at 0.031 mg ml-1. While the least activity, albeit still a strong inhibitor, was observed from the Yzerfontein extracts with an MIC value of 0.5 mg ml-1. Generally comparison between the two years revealed that samples collected in 2011 were more potent than those in 2009, possibly due to prolonged storage that may have resulted in chemical decomposition. As the Stellenbosch population had shown the best activity as well as a relatively different chemical profile, leaves from these plants were then introduced into tissue culture conditions. Leaf explants were placed on solid plant growth regulator (PGR)-free Murashige and Skoog media and that supplemented with hormones in various combinations. (two concentrations of benzyl adenine (BA) utilized individually and in combination with naphthalene acetic acid (NAA) 4.4 and 8.8, while for NAA 0.27, 2.7 and 5.4.) Namely: 4.4 BA, 8.8 BA, 0.27 NAA: 4.4 BA, 2.7 NAA: 4.4 BA, 5.4 NAA: 4.4 BA, 2.7 NAA: 8.8 BA and 5.4 NAA: 8.8 BA. The PGR combinations did not induce shooting nor rooting, only callus on PGR-free MS media. Antifungal activity of the callus extract was in the same range as the whole plant extracts from which the leaf explants were harvested from, showing no ‘loss’ of activity after introduction to tissue culture conditions. Metabolite profiles using LC-MS, however, did reveal qualitative and quantitative differences, though they appear to not have any bearing on the activity. A bioassay-guided fractionation was then conducted on samples collected from Stellenbosch. This led to the identification of carnosol and carnosic acid being involved in the anti-Fusarium activity of S africana-lutea. A combinational study revealed no synergistic activity of the two compounds against four Fusarium test strains, with fractional inhibitory concentration (FIC) values of 1.5 and 3.0. Antifungal activity of carnosol and carnosic acid was observed to be in the same range (strong inhibitor) as was the callus and whole plant extracts. The study showed variation in population chemotypes and identified two compounds that are involved in S. africana-lutea activity against Fusarium species. It also provided a tissue culture system onto which mass production of the two bioactives may be achieved from, in the development of new fungicides. / AFRIKAANSE OPSOMMING: Salvia Africana lutea is een van die 26 Salie spesies wat inheems aan Suid-Afrika is uit ‘n totaal van 900 spesies wêreldwyd. Die genus Salvia hoort tot die Lamiaceae familie. S. africana lutea word geklassifiseer as ‘n “breë spektrum geneesmiddel”, en medisinaal as belangrik geag tussen die ander salie spesies, want volgens verslag word dit gebruik vir die verligting van hoes en verkoues tot selfs ginekologiese ongesteldhede. Definitiewe biologiese aktiwiteit eksperimente het anti-mikrobiese, antikanker en anti-oksidant aktiwiteite aan die lig gebring. Plant sekondêre metaboliete het fundamenteel ‘n baie sterk verband met die fitochemiese aktiwiteite van ‘n plant. Gevolglik affekteer die omgewing indirek die fitochemiese eienskappe, want dit beïnvloed die variasie in die plant metaboloom deur die interaksies van die plant met die omgewing. In vyf streke binne die Wes-Kaap van Suid-Afrika waar S. africana lutea bevolkings voorkom, is steekproewe gedoen en chemotipes en bioaktiwiteit getoets. Vier bevolkings was wild-groeiende bevolkings in beskermde areas, naamlik; Brackenfell, Koeberg, Silwerstroomstrand en Yzerfontein, terwyl die vyfde uit ‘n tuingroeiende bevolking in Stellenbosch geneem is. Deur gas chromatografie gekoppel met massa spektrometrie te gebruik, is primêre samestellings soos monosakkariede, karboksielsure en vetsure gevind. Variasies van samestellings wat met 80% sekerheid geïdentifiseer is, is oorkruis met al die bevolkings vergelyk. Die Stellenbosch bevolking het ‘n paar samestellings geopenbaar wat nie aanwesig was in die ander vier terreine nie. Hierdie samestellings was: propanoësuur, erythroniese suur, 2-keto-1-glukoniese suur en 1,3-dibromobicyclon. Verder het hierdie bevolking geen xylitol gehad nie en dit is in al vier die ander bevolkings gevind. Verdere studies was gedoen met die gebruik van vloeibare chromatografie gekoppel met massa spektrometrie (LC-MS) sowel as kern magnetiese resonansie (1H NMR). Chemiese profiele het hoë variasies getoon, en dus deur te fokus op die aromatiese samestelling streke, het die Stellenbosch terrein duidelik merkbare verskille en punte op die PLS-DA aangetoon. Met die koppeling van NMR data met LC-MS data, is daar gevind dat onderskeidende punte van die NMR PLS-DA wat gegroepeer is met retensie tye die skeiding van die Stellenbosch terrein van ander terreine gedryf het. Dit het onweerlegbaar bewys dat daar variasie binne die vyf bevolkings voorkom en dat Stellenbosch die mees noemenswaardige chemotipe variasie het. Dit blyk uit die anti-mikrobiese eksperimente dat aktiwiteit teen Fusarium heel nuwe belangstelling wek. Fusarium is ‘n plant-patogeniese spesie wat groot landbou verliese veroorsaak veral in die mielie gewasse, een van die stapelvoedsels van die Afrika kontinent. Dit produseer ook mikotoksiene in aangetaste gewasse en hierdie kan lei tot die ontstaan van kankers wanneer besmette voedsel op groot skaal verbruik word. Vier hoë-mikotoksien produserende swamlyne van twee spesies, naamlik F. verticillioides (MRC 826 en MRC 8267) en F. proliferatum (MRC 7140 en MRC 6809) is gebruik in alle in vitro anti-swam ondersoeke in hierdie studie. Die eerste analise het dichloromethan: methanol (1:1 v/v) ongesuiwerde plant ekstrakte bevat van die vyf bevolkings: Stellenbosch, Brackenfell, Koeberg, Silwerstroomstrand en Yzerfontein, geneem gedurende 2009 en 2011. Al hierdie toets monsters het goeie aktiwiteit getoon waar die minimum beperkende konsentrasie (MIC) waardes van 0.031 mg ml-1 tot 0.5 mg ml-1 gevarieer het. Waardes laer as laasgenoemde word beskou as sterk inhibeerders. Die Stellenbosch ekstrakte was die mees aktief vir albei jare hierbo genoem, met die beste aktiwiteit teen F. verticillioides MRC 8267 en MCR 826 by 0.031 mg ml-1. Die minste aktiwiteit (hoewel nog ‘n sterk inhibeerder) waargeneem was van die Yzerfontein ekstrakte, met ‘n MIC waarde van 0.5 mg ml-1. Oor die algemeen het ‘n vergelyking tussen die twee jare aangetoon dat die monsters wat in 2011 versamel is veel sterker was dan dié van 2009, moontlik te wyte aan ‘n verlengde bewaringstyd wat moontlik ‘n chemiese dekomposisie ten gevolge gehad het. Omdat die Stellenbosch bevolking die beste aktiwiteit getoon het sowel as ‘n relatief afwykende chemiese profiel, is blare van hierdie plante toe bekendgestel aan weefselkultuur kondisies. Blaar eksplante is op soliede hormoonvrye Murashige en Skoog media geplaas en dit is aangevul met sintetiese auksien Naftaleen asynsuur (NAA) en sitokien Bensiel adenien (BA) individueel en in verskillende kombinasies. Geen wortels of uitloopsels is waargeneem in al die hormoon kombinasies nie maar in die hormoonvrye media het daar egter Kallus in twintig persent van die eksplante voorgekom. Kallus is toe as subkultuur van hormoonvrye MS media gekweek en saamgevoeg en dichloromethan: methanol (1:1v/v) ekstrakte is getoets teen die volgende Fusarium swamlyne MRC 826; MRC 8267; MRC 7140 en MRC 6809. MIC waardes het sterk inhiberende eienskappe getoon met die laagste waarde as 0.025 mg ml-1 teen drie swamlyne: MRC 1740, MRC 8267 en MRC 826, en die hoogste was 0.25 mg ml-1 na 48 uur. Die minimum inhiberende konsentrasie waardes het gestyg na 0.5mg ml-1 na 60 uur, wat ‘n fungistatiese aksie getoon het. Maar van 60 tot 92 uu het waardes egter ‘n swamdodende aksie aangetoon met geen verandering van 0.5mg ml-1 nie. In die identifisering van die bioaktiewe komponente, is die ekstraksie van Stellenbosch se bevolking in dichloromethan: methanol (1:1 v/v) uitgevoer, en met gebruik van vyftig gram van die ekstrak is bioanalise-geleide fraksionering gedoen deur gebruik van ‘n VersaFlash®. Die mees aktiewe fraksie is verder gefraksioneer deur die gebruik van ‘n konvensionele silikajel kolom. Aktiewe fraksies is getoets deur LC-MS te gebruik, en twee verbindings, carnosol en carnosic suur, is geïdentifiseer. Voorbereidende TLC is gebruik om identiteit te bevestig, want fraksies was naas die kommersiele standaarde van die twee verbindings getoets. Sinergistiese aktiwiteit van die twee samestellings is ondersoek deur ‘n antiswam ontleding teen die vier swamlyne uit te voer. Hierdie studie het dus die veronderstelde bestaan van verskillende chemotipes tussen die bevolkings waarvan voorbeelde geneem is, bekend gestel. Veral die Stellenbosch se bevolking het die meeste verskil, heel moontlik omrede die verlengde en hoër versteurings deur die nabyheid van mense. Plant– omgewing interaksies speel ‘n belangrike rol in die metaboloom van plante, wat dan indirek hul eienskappe verander, en in hierdie geval die antiswam aktiwiteit. Die tuingroeiende bevolking was die mees aktief, heel moontlik omrede hierdie aspek. Nietemin was geen bioaktiwiteit verloor waar die mees kragtige bevolking met weefsel kultuur kondisies in aanraking gebring is nie. Dus is dit ideaal vir kommersialisering. Een nuwe belangrike bevinding was die carnosol en carnosic suur wat twee welbekende samestellings is wat meesal geassosieer is met Rosmarinus officinalis en gedokumenteer is vir antioksidant aktiwiteit. Hier dui laasgenoemde samestellings ‘n antiswam aktiwiteit aan teen die getoetste Fusarium swamlyne. Met ‘n gevestigde weefsel kultuur sisteem alreeds in plek, voorsien dit ‘n beginpunt vir die bestudering van hoe hierdie bioaktiewe komponente in massa geproduseer kan word in die ontwikkeling van nuwe swamdodende produkte.

O cultivo da chia (Salvia hispânica L.) tem apresentado enorme importância em diversos países, porém, são poucas as pesquisas em relação ao manejo agronômico que permitam auxiliar sobre as exigências da cultura. No Brasil a cultura está sendo recentemente adaptada e tem se deparado com alguns fatores limitantes à obtenção de elevadas produtividades, dentre eles a adubação nitrogenada e fosfata. Desta forma objetivou-se com este trabalho avaliar a combinação de doses de nitrogênio e fósforo na cultura de chia, em solos de cerrado, no sul do estado do Tocantins. Os experimentos foram conduzidos em campo na Estação Experimental da Universidade Federal de Tocantins na safra dos anos 2014/2015 e 2015/2016. O delineamento utilizado foi blocos ao acaso, num esquema fatorial 5x5 com três repetições na safra 2014/2015 e quatro repetições na safra 2015/2016, ambos constituídos por a combinação de cinco níveis de adubação fosfatada (0, 30, 60, 90 e 120 kg ha-1) e cinco níveis de adubação nitrogenada (0, 30, 60, 90 e 120 kg ha-1). As características avaliadas foram altura da planta, altura da haste superior, diâmetro do caule, número de cachos, comprimento do cacho e produtividade de grãos. Concluiu-se as doses de 60 e 120 kg ha-1 de nitrogênio 10 proporcionam a maior altura de planta, altura da haste superior e diâmetro do caule; baixas doses de 30 e 60 kg ha-1 de fósforo influenciaram de forma positiva no maior desenvolvimento e produtividade na cultura da chia. Já uma combinação de 30 kg ha-1 de N e 60 kg ha-1 de P favoreceu a maior comprimento de cachos. A produtividade de grãos máxima encontrada nas doses isolada de N e P foi superior de 157 kg ha-1. / The cultivation of chia (Salvia hispânica L.) has presented enormous importance in many countries, because there is little research in relation to the agronomic management to allow aid on the requirements of the culture. In Brazil, the culture being recently adapted and has met with some limiting factors to obtain high yields, including nitrogen and phosphate fertilization. Thus, the aim of this study was to evaluate the combination of nitrogen and phosphorus fertilization in chia culture, in Cerrado soils, in the South of Tocantins. The experiments were conducted in the field at the Experimental Station of the Federal University of Tocantins in the harvests in the years 2014/2015 and 2015/2016. The design was a randomized block design, in a 5x5 factorial design with three replications in the harvests 2014/2015 and four replications in the harvests 2015/2016, both consist by the combination of five levels of phosphorus fertilizer (0, 30, 60, 90 e 120 kg ha-1) and five levels of nitrogen fertilizer (0, 30, 60, 90 e 120 kg ha-1). The characteristics evaluated were plant height, upper stem height, stem diameter, bunches number, bunch length and grain yield It was concluded that these doses of 60 and 120 kg ha-1 of nitrogen provide the greatest plant height, upper stem height and stem diameter and low doses of 30 and 60 kg ha-1 of phosphorus influenced positively in the most development and productivity in the culture of chia.The combination of 30 kg ha-1 of N and 60 kg ha-1 P was sufficient to promote the biggest issue number of bunches in the chia plant. The maximum grain yield found in the isolated doses of N and P was superior to 157 kg ha-1.

Tse Pui Shan. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2003. / Includes bibliographical references (leaves [191]-216). / Abstracts in English and Chinese. / ACKNOWLEDGEMENTS --- p.I / ABBREVIATIONS --- p.III / ABSTRACT --- p.VIII / 摘要 --- p.X / PUBLICATIONS --- p.XII / TABLE OF CONTENTS --- p.XIII / Chapter CHAPTER 1: --- GENERAL INTRODUCTION / Chapter 1.1 --- Human Immune System and Cancer --- p.1 / Chapter 1.1.1 --- Brief Introduction of the Human Immune System --- p.1 / Chapter 1.1.2 --- Prevalence of Cancer in Hong Kong --- p.4 / Chapter 1.1.3 --- The Role of the Immune System in Tumorigenesis --- p.4 / Chapter 1.1.4 --- Cancer Treatment --- p.5 / Chapter 1.1.5 --- Cancer Prevention --- p.5 / Chapter 1.2 --- Mushroom Polysaccharides --- p.6 / Chapter 1.2.1 --- General Aspects of Mushroom Polysaccharides --- p.6 / Chapter 1.2.2 --- Structure of Mushroom Polysaccharides --- p.9 / Chapter 1.2.2.1 --- Beta (P)-D-glucans --- p.9 / Chapter 1.2.2.2 --- Heteroglucans and Protein-bound Polysaccharides --- p.10 / Chapter 1.2.2.3 --- Structure-Function Interactions of Polysaccharides --- p.12 / Chapter 1.2.3 --- Molecular Interactions of Polysaccharides --- p.14 / Chapter 1.2.4 --- Biological Activities of Polysaccharides --- p.15 / Chapter 1.2.4.1 --- Anti-tumor Activities of Polysaccharides --- p.15 / Chapter 1.2.4.2 --- Immunomodulatory Activities of Polysaccharides --- p.16 / Chapter 1.3 --- Yun Zhi (Coriolus versicolor) --- p.17 / Chapter 1.3.1 --- General Features of Yun Zhi --- p.17 / Chapter 1.3.2 --- Traditional Uses of Yun Zhi --- p.20 / Chapter 1.3.3 --- Active Ingredients of Yun Zhi --- p.20 / Chapter 1.3.3.1 --- "Origin, Properties and Composition of PSK" --- p.21 / Chapter 1.3.3.2 --- "Origin, Properties and Composition of PSP" --- p.22 / Chapter 1.3.4 --- Pharmacological Actions of PSP and PSK --- p.25 / Chapter 1.3.4.1 --- Immunomodulatory Activities --- p.25 / Chapter 1.3.4.2 --- Anti-tumor Activities --- p.32 / Chapter 1.3.4.2 --- Antiviral and Antimicrobial Activities --- p.35 / Chapter 1.3.4.3 --- Antioxidant Activities --- p.36 / Chapter 1.3.5 --- Human Clinical Studies on Yun Zhi --- p.36 / Chapter 1.3.6 --- Toxicology of Yun Zhi --- p.42 / Chapter 1.4 --- Danshen (Salvia miltiorrhiza) --- p.43 / Chapter 1.4.1 --- General Features of Danshen --- p.43 / Chapter 1.4.2 --- Traditional Uses of Danshen --- p.46 / Chapter 1.4.3 --- Active Ingredients of Danshen --- p.47 / Chapter 1.4.4 --- Pharmacological Actions of Danshen --- p.50 / Chapter 1.4.4.1 --- Cardiovascular Effects --- p.50 / Chapter 1.4.4.2 --- Scavenging Effects on Free Radicals --- p.52 / Chapter 1.4.4.3 --- Hepatoprotective Effects --- p.54 / Chapter 1.4.4.4 --- Anti-tumor Effects --- p.56 / Chapter 1.4.4.5 --- Renal Protective Effects --- p.56 / Chapter 1.4.5 --- Human Clinical Studies --- p.57 / Chapter 1.4.6 --- Toxicity of Danshen --- p.59 / Chapter 1.5 --- Aims and Scopes of This Investigation --- p.60 / Chapter CHAPTER 2: --- MATERIALS AND METHODS / Chapter 2.1 --- Normal Subjects --- p.62 / Chapter 2.1.1 --- Inclusion and Exclusion Criteria of Recruitment --- p.62 / Chapter 2.1.2 --- Study Design and Procedure --- p.63 / Chapter 2.1.3 --- Treatment and Blinding --- p.65 / Chapter 2.1.4 --- Blood Sampling --- p.66 / Chapter 2.1.5 --- Blood Processing for Assessment of Immunological Functions --- p.67 / Chapter 2.2 --- Materials --- p.69 / Chapter 2.2.1 --- Endotoxin Assay --- p.69 / Chapter 2.2.2 --- Reagents for Whole Blood Assay --- p.69 / Chapter 2.2.2.1 --- Plain RPMI 1640 Medium --- p.69 / Chapter 2.2.2.2 --- Phosphate-Buffered Saline (PBS) --- p.69 / Chapter 2.2.2.3 --- Mitogens --- p.70 / Chapter 2.2.3 --- Reagents for Total RNA Extraction --- p.70 / Chapter 2.2.3.1 --- Ficoll-Paque Density Gradient Solution --- p.70 / Chapter 2.2.3.2 --- RNA Extraction Kit --- p.70 / Chapter 2.2.3.3 --- RNase-Free DNase Set --- p.71 / Chapter 2.2.3.4 --- β-Mercaptoethanol (β-ME) Solution --- p.71 / Chapter 2.2.4 --- Reagents for Flow Cytometric Analysis of T/B/NK Cell Ratios --- p.71 / Chapter 2.2.4.1 --- MultiTEST IMK Kit with TruCOUNT Tubes --- p.71 / Chapter 2.2.4.2 --- FACSFlo´wёØ Sheath Fluid --- p.74 / Chapter 2.2.4.3 --- CaliBRITE 3 and APC Beads --- p.74 / Chapter 2.2.5 --- Immunoassay Kits for Measuring Cytokines Level --- p.75 / Chapter 2.2.5.1 --- Enzyme-linked Immunosorbent Assay (ELISA) Kits of Cytokines --- p.75 / Chapter 2.2.5.2 --- Human Thl/Th2 Cytokine Cytometric Bead Array (CBA) Kit-II --- p.75 / Chapter 2.2.6 --- Reagents and Buffers for Gel Electrophoresis --- p.78 / Chapter 2.2.6.1 --- Ethidium Bromide (EtBr) --- p.78 / Chapter 2.2.6.2 --- Gel Loading Solution (5X) --- p.78 / Chapter 2.2.6.3 --- Tris-Acetate-EDTA (TAE) Buffer --- p.78 / Chapter 2.2.6.4 --- Agarose Gel --- p.78 / Chapter 2.2.6.5 --- 100 base pair DNA Ladder --- p.79 / Chapter 2.2.7 --- Kits and Reagents for Messenger RNA (mRNA) Expression Array --- p.79 / Chapter 2.2.7.1 --- Human Inflammatory Cytokine/Receptor GEArraýёØ Q Series Kit --- p.79 / Chapter 2.2.7.2 --- Deoxynucleoside Triphosphates (dNTPs) --- p.84 / Chapter 2.2.7.3 --- Moloney Murine Leukemia Virus Reverse Transcriptase (M-MLVRT) --- p.84 / Chapter 2.2.7.4 --- Rnasin Ribonuclease Inhibitor --- p.84 / Chapter 2.2.7.5 --- Biotin-16-2'-deoxy-uridine-5'-triphosphate (Biotin-16-dUTP) --- p.85 / Chapter 2.2.7.6 --- Salmon Sperm DNA Solution --- p.85 / Chapter 2.2.7.7 --- 100 % Sodium Dodecyl Sulfate (SDS) Solution --- p.86 / Chapter 2.2.7.8 --- 20X SSC --- p.86 / Chapter 2.2.7.9 --- ECL Films (Hyperfilm 226}0ёØ ECL 226}0ёØ) --- p.86 / Chapter 2.3 --- Methods / Chapter 2.3.1 --- Endotoxin Assay --- p.87 / Chapter 2.3.2 --- Whole Blood Assay (WBA) --- p.88 / Chapter 2.3.3 --- Isolation and Preparation of Plasma and Peripheral Blood Mononuclear Cells (PBMC) from EDTA Blood --- p.88 / Chapter 2.3.4 --- Total RNA extraction --- p.89 / Chapter 2.3.5 --- Flow Cytometric Analysis of T/B/NK Cell Ratios --- p.90 / Chapter 2.3.6 --- Immunoassays of Plasma Samples or Culture Supernatant in WBA --- p.92 / Chapter 2.3.6.1 --- Enzyme-linked Immunosorbent Assay (ELISA) --- p.92 / Chapter 2.3.6.2 --- Human Thl/Th2 Cytokine Cytometric Bead Assay (CBA) --- p.93 / Chapter 2.3.7 --- mRNA Expression Study --- p.94 / Chapter 2.3.7.1 --- Agarose Gel Electrophoresis --- p.94 / Chapter 2.3.7.2 --- cDNA Expression Array Analysis --- p.95 / Chapter 2.3.8 --- Statistical Analysis --- p.96 / Chapter CHAPTER 3: --- ENDOTOXIN LEVEL OF YUN ZHI-DANSHEN CAPSULES & SAFETY MEASURES ON STUDY POPULATION IN THE CLINICAL TRIAL / Chapter 3.1 --- Introduction --- p.98 / Chapter 3.2 --- Results --- p.101 / Chapter 3.2.1 --- Endotoxin Level of the Yun Zhi and Danshen Active Capsule --- p.101 / Chapter 3.2.2 --- Study Population --- p.103 / Chapter 3.2.3 --- Dropout Cases --- p.103 / Chapter 3.2.4 --- Safety Parameters --- p.104 / Chapter 3.2.5 --- Compliance Rates --- p.104 / Chapter 3.3 --- Discussion --- p.109 / Chapter CHAPTER 4: --- FLOW CYTOMETRIC ANALYSIS OF T/B/NK CELL RATIOS OF HEALTHY SUBJECTS TAKING YUN ZHI-DANSHEN CAPSULES / Chapter 4.1 --- Introduction --- p.112 / Chapter 4.2 --- Results --- p.118 / Chapter 4.2.1 --- The Percentage and Absolute Count of T Lymphocytes (CD3+) --- p.118 / Chapter 4.2.2 --- The Percentage and Absolute Count of T Helper (TH) Lymphocytes (CD3+ CD4+) --- p.121 / Chapter 4.2.3 --- The Percentage and Absolute Count of Cytotoxic T (CTL) and T Suppressor (Ts) Lymphocytes (CD3+ CD8+) --- p.124 / Chapter 4.2.4 --- The Ratio of T Helper Lymphocytes (CD3+ CD4+) and Cytotoxic T (CTL) and T Suppressor (Ts) Lymphocyes (CD3+ CD8+) --- p.127 / Chapter 4.2.5 --- The Percentage and Absolute Count of B Lymphocytes (CD19+) --- p.129 / Chapter 4.2.6 --- The Percentage and Absolute Count of NK Lymphocytes (CD3- CD 16+ and/or CD56+) --- p.132 / Chapter 4.2.7 --- The Absolute Count of Lymphocytes (CD45+) --- p.135 / Chapter 4.3 --- Discussion --- p.138 / Chapter CHAPTER 5: --- PLASMA CONCENTRATION OF SOLUBLE CYTOKINE RECEPTOR AND EX VIVO CYTOKINE PRODUCTION OF HEALTHY SUBJECTS TAKING YUN ZHI-DANSHEN CAPSULES / Chapter 5.1 --- Introduction --- p.142 / Chapter 5.2 --- Results --- p.147 / Chapter 5.2.1 --- Plasma Concentration of Soluble IL-2 Receptor --- p.147 / Chapter 5.2.2 --- Ex vivo Cytokine Production --- p.147 / Chapter 5.2.3 --- Mitogen Induced IL-6 Production --- p.150 / Chapter 5.2.4 --- Mitogen Induced IFN- γ Production --- p.150 / Chapter 5.2.5 --- Mitogen Induced TNF- a Production --- p.153 / Chapter 5.2.6 --- Mitogen Induced IL-10 Production --- p.153 / Chapter 5.3 --- Discussion --- p.156 / Chapter CHAPTER 6: --- "GENE EXPRESSION OF CYTOKINES, CHEMOKINES AND RECEPTORS OF PBMC OF HEALTHY SUBJECTS TAKING YUN ZHI- DANSHEN CAPSULES" / Chapter 6.1 --- Introduction --- p.162 / Chapter 6.2 --- Results --- p.165 / Chapter 6.2.1 --- Gene Expression of IL-2 Receptor β chain --- p.165 / Chapter 6.2.2 --- Gene Expression of IL-2 Receptor γ chain --- p.165 / Chapter 6.2.3 --- Gene Expression of IL-6 Receptor --- p.166 / Chapter 6.2.4 --- "Gene Expression of Other Cytokines, Chemokines and Receptors" --- p.169 / Chapter 6.3 --- Discussion --- p.172 / Chapter CHAPTER 7: --- CONCLUDING REMARKS AND FUTURE / PERSPECTIVES --- p.176 / APPENDICES --- p.184 / REFERENCES --- p.192

Ng Chau Wah Stephen. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2003. / Includes bibliographical references (leaves 120-135). / Abstracts in English and Chinese. / Abstract --- p.i / Acknowledgements --- p.viii / Publications --- p.ix / Abbreviations --- p.x / Chapter Chapter 1 --- Introduction / Chapter 1.1 --- Traditional Chinese Medicine --- p.1 / Chapter 1.2 --- Danshen --- p.7 / Chapter 1.2.1 --- Chemical constituents of Danshen --- p.7 / Chapter 1.2.2 --- Pharmacology of Danshen --- p.10 / Chapter 1.3 --- Vascular system --- p.13 / Chapter 1.3.1 --- Physiology of blood vessels --- p.13 / Chapter 1.3.2 --- Vascular smooth muscle contraction --- p.14 / Chapter 1.3.3 --- Mechanism of vascular smooth muscle contraction --- p.15 / Chapter 1.3.3.1 --- Adrenoceptor in vascular system --- p.19 / Chapter 1.3.3.2 --- Muscarinic receptor in vascular system --- p.20 / Chapter 1.3.3.3 --- Synthesis and release of Nitric Oxide (NO)in vascular system --- p.22 / Chapter 1.3.3.4 --- Synthesis and release of postanoidsin vascular system --- p.25 / Chapter 1.3.3.5 --- Synthesis and release of histaminein vascular system --- p.28 / Chapter 1.3.3.6 --- Synthesis and release of Calcitonin gene-related peptide in vascular system --- p.29 / Chapter 1.4 --- Aims of the studies --- p.33 / Chapter Chapter 2 --- Materials and methods / Chapter 2.1 --- Materials --- p.35 / Chapter 2.2 --- Methods - General procedures --- p.35 / Chapter 2.2.1 --- Preparations of drug solutions --- p.35 / Chapter 2.2.2 --- Animals used and anaesthetization --- p.36 / Chapter 2.2.3 --- Cannulation of carotid artery and jugular vein --- p.37 / Chapter 2.2.4 --- Blood pressure measurement --- p.37 / Chapter 2.2.5 --- Knee joint denervation --- p.38 / Chapter 2.2.6 --- Knee joint blood flow measurement --- p.39 / Chapter 2.3 --- Methods - Specific procedures --- p.41 / Chapter 2.3.1 --- Validation of Laser Doppler Imaging (LDI) measurements --- p.41 / Chapter 2.3.2 --- Actions of topical administration of Danshen --- p.42 / Chapter 2.3.2.1 --- Studies of the mechanism(s) of action of Danshen --- p.43 / Chapter 2.3.2.2 --- Investigation for α-adrenoceptor antagonist activity --- p.44 / Chapter 2.3.2.3 --- Investigation for neural involvement --- p.44 / Chapter 2.3.3 --- Actions of intravenous administration of Danshen --- p.45 / Chapter 2.3.4 --- Data analysis --- p.45 / Chapter Chapter 3 --- Results / Chapter 3.1 --- Validation of LDI measurement --- p.47 / Chapter 3.2 --- Actions of intravenous administration of Danshen --- p.53 / Chapter 3.3 --- Actions of topical administration of Danshen --- p.53 / Chapter 3.4 --- Muscarinic receptor antagonist on Danshen --- p.61 / Chapter 3.5 --- β-adrenoceptor antagonist on Danshen --- p.67 / Chapter 3.6 --- Danshen on α-adrenoceptor agonist-induced vasoconstriction --- p.74 / Chapter 3.7 --- Nitric oxide synthase inhibitor on Danshen --- p.79 / Chapter 3.8 --- Cyclo-oxygenase (COX) inhibitor on Danshen --- p.83 / Chapter 3.9 --- Histamine receptor antagonists on Danshen --- p.87 / Chapter 3.10 --- CGRP receptor antagonist on Danshen --- p.92 / Chapter 3.11 --- Effect of denervation on Danshen --- p.92 / Chapter Chapter 4 --- Discussion --- p.100 / Reference --- p.120

Submitted by Maria Beatriz Vieira (mbeatriz.vieira@gmail.com) on 2017-03-30T12:42:14Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) dissertacao_maria_johana_gonzalez_vera.pdf: 903923 bytes, checksum: 2b38e1d63ed072f664036343cb26b7d7 (MD5) / Approved for entry into archive by Aline Batista (alinehb.ufpel@gmail.com) on 2017-04-05T18:01:53Z (GMT) No. of bitstreams: 2 dissertacao_maria_johana_gonzalez_vera.pdf: 903923 bytes, checksum: 2b38e1d63ed072f664036343cb26b7d7 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Aline Batista (alinehb.ufpel@gmail.com) on 2017-04-05T18:23:27Z (GMT) No. of bitstreams: 2 dissertacao_maria_johana_gonzalez_vera.pdf: 903923 bytes, checksum: 2b38e1d63ed072f664036343cb26b7d7 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2017-04-05T18:23:37Z (GMT). No. of bitstreams: 2 dissertacao_maria_johana_gonzalez_vera.pdf: 903923 bytes, checksum: 2b38e1d63ed072f664036343cb26b7d7 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2015-03-30 / Sem bolsa / A Salvia hispanica L. pertence à família Lamiaceae, é nativa do México e conhecida popularmente como "chia”. Os frutos, erroneamente chamados de sementes, possuem diversos usos medicinais, culinário, artístico e religioso que tornaram a cultura importante, especialmente no aspecto nutricional em razão do alto teor de ácidos graxos ômega-3. Atualmente, inexistem metodologias para avaliar a germinação das sementes da espécie S. hispanica nas regras da ISTA (International Seed Testing Association) e nas Regras para Análise de Sementes, sendo utilizadas como referência outras espécies do mesmo gênero. O objetivo de estabelecer procedimentos metodológicos para realização do teste de germinação em sementes de Salvia hispanica L. Foram utilizados sete lotes de sementes de chia que foram submetidas a seis regimes de temperatura (20 ºC; 25 ºC, 30 ºC; 20-30 ºC; 15-35 ºC; 20-35 ºC) na presença e na ausência de luz. Utilizou-se semeadura em gerbox, utilizando-se substrato sobre papel. Foram realizadas contagens diárias para determinar as épocas adequadas para realizar as avaliações. Avaliou-se Germinação, Primeira contagem da germinação, índice de velocidade de germinação, velocidade de germinação, coeficiente de velocidade de emergência e emergência de plântulas. Os resultados permitem concluir que as sementes de S. hispanica L. apresentam comportamento indiferente à luz, as contagens no teste de germinação devem ser realizadas aos quatro e sete dias após a semeadura e que as temperaturas mais adequadas para realização do teste de germinação são constantes de 20 ºC e alternadas de 20-30 ºC. / Salvia hispanica belongs to the Lamiaceae family, is native to Mexico and popularly known as "chia". The fruits wrongly called seeds, have many medicinal, culinary, artistic and religious uses that have made important this crop, especially in the nutritional aspect due to the high content of fatty acids like omega-3. Currently there are no analysis methodologies to evaluate the germination of S. hispanica seeds in the ISTA (International Seed Testing Association) rules or RAS (Regras para Análise de Sementes) rules, being used as a reference other species of the same genus. The objective of this study was to establish methodological standards for conducting the germination test in S. hispanica seed. Seven lots of chia seeds were used that were subjected to six different temperatures (20 ° C, 25 ° C, 30 ° C; 20-30 ° C; 15-35ºC, 20-35ºC) in the presence and absence of light. Seeding was made in gerbox boxes, using the substrate on paper. Daily counts were made to determine the appropriate times to perform the evaluations. Germination, First count of germination, IVG, GV, CVG and Emergency were evaluated. The results showed that the seeds of S. hispanica L. feature light the indifferent behavior , the counts in the germination test should be conducted at four and seven days after sowing and the most suitable temperature for performing the germination test are constant 20 ° C and 20-30 ° C alternating.

Salvia genus is a widely cultivated genus and used in medicine for various purposes as having antimicrobial, antioxidant, anticarcinogen and anti-inflammatory features. In this study the aim was to investigate phenolic composition of Salvia absconditiflora and understand the possible effects of those constituents in cancer related drug metabolizing enzymes. Salvia absconditiflora showed 80,43 % Radical Scavenging Activity against DPPH radical. Total flavonoid content was found as one third of total phenolic content. Presence of important phenolic acids and flavonoids such as caffeic acid, luteolin, coumaric acid are validated with LC-MS/MS analysis. Cytotoxicity of Salvia absconditiflora treatment on MCF-7 and MDA-MB-231 breast cancer cell lines were investigated through XTT and TBE assays both dose and time dependent manner. Cell proliferation was inhibited 50 % by different IC50 values calculated in different assays and different time intervals. This suggests that two breast cancer cell lines response in a different way to cytotoxic treatments. Cancer related drug metabolizing enzyme gene modulations were investigated with qRT-PCR. CYP1A1 and CYP1B1 were upregulated in MCF-7 but down-regulated in MDA-MB-231 cells in response to Salvia absconditiflora treatment.

Salvia genus is a widely cultivated genus and used in medicine for various purposes as having antimicrobial, antioxidant, anticarcinogen and anti-inflammatory features. In this study the aim was to investigate phenolic composition of Salvia absconditiflora and understand the possible effects of those constituents in cancer related drug metabolizing enzymes. Salvia absconditiflora showed 80,43 % Radical Scavenging Activity against DPPH radical. Total flavonoid content was found as one third of total phenolic content. Presence of important phenolic acids and flavonoids such as caffeic acid, luteolin, coumaric acid are validated with LC-MS/MS analysis. Cytotoxicity of Salvia absconditiflora treatment on MCF-7 and MDA-MB-231 breast cancer cell lines were investigated through XTT and TBE assays both dose and time dependent manner. Cell proliferation was inhibited 50 % by different IC50 values calculated in different assays and different time intervals. This suggests that two breast cancer cell lines response in a different way to cytotoxic treatments. Cancer related drug metabolizing enzyme gene modulations were investigated with qRT-PCR. CYP1A1 and CYP1B1 were up-regulated in MCF-7 but down-regulated in MDA-MB-231 cells in response to Salvia absconditiflora treatment.

A revision of the Mediterranean and the Aegean geographic regions of Turkey of Salvia L. (Labiatae) is presented. According to the results, the study area has 60 taxa, 32 of which are endemic (mainly local endemics), 5 of which are non-endemic rare, and the remaining 23 taxa are widely distributed. The rate of endemism is 53% in the area. At the beginning of the thesis, history and worldwide distribution of the family and the genus are briefly given. Phylogenetic studies and infrageneric grouping of the genus are discussed. As well as, recent studies on the genus such as taxonomical, morphological, micromorphological and karyological are provided. Geographic, phytogeographic, topographic, and climatic features of the area are explained. Since 2005, about 2000 specimens have been collected and examined as the research materials. In addition, a large number of specimens have been seen and studied in ANK, AEF, BM, E, ERCIYES, G, GAZI, HUB, K, KNYA, LE, ISTE and ISTF herbaria. Digital photos of some specimens including type specimens have been seen from B, W and A herbaria. In the field when the specimens were detected geographical location (including GPS), habitat, phenological data and relevant field observations were all recorded. In the study, comparative morphological characters and their variations are discussed Anatomical features, such as root, stem, leaf and petiole, micromorphological properties, such as trichome, pollen and nutlet, of the species are comprehensively studied and their taxonomic implications are discussed. According to field studies and literature surveys, endemism, pyhtogeography, habitat, phenology, chorology, new distribution areas and conservation status of the species are determined. Infrageneric delimitation of the species using multivariety analysis is performed. In addition, first sectional key and sectional grouping of the species are made. As well as, identification key of the species is given. At the end of the thesis, expanded and corrected description, address, photographs and some notes on taxonomy of the species are given. The taxonomic studies revealed one new species, Salvia marashica, two new varieties, S. sericeo-tomentosa var. hatayica, S. cadmica var. bozkiriensis, one new combination, S. aucheri subsp. canescens and one new record, S. viscosa. As well as, S. sericeo-tomentosa and S. quezelii are rediscovered.

This study aims to revise the genus Salvia L. in East and Southeast Anatolia in Turkey on the basis of macromorphological, anatomical, palynological, mericarp micro-morphological, ecological and numerical analysis. Towards achieving this goal, about 2500 specimens of Salvia were collected and examined during extensive field studies between July 2005 and June 2009. The materials collected by other researchers either from Turkey or abroad were also investigated. The results of the taxonomic revision show that the study area includes 59 taxa, 24 (40.7%) of which are endemic and the remaining 35 (59.3%) are non-endemic. Salvia siiirtica is described as new to science. S. macrosiphon is described as a new record for Turkey and S. cerino-pruinosa and S. pseudeuphratica are re-evaluated as valid species as well as S. ballsiana is rediscovered. Macromorphological characters that were considered to have taxonomically diagnostic value are investigated and their possible variations are discussed. Habit, stem, leaf, inflorescence, bract, calyx, corolla and stamen properties are compared at infrageneric and species level. The first comprehensive evaluation of the systematic value of anatomy, palynology and mericarp micromorphology of Salvia are presented. Some characters, such as row numbers of ray cells in the root, type of the leaf blade structure, shape of cross section of the petiole, shape and number of median vascular bundles, pollen size, size of mericarps and diameter of abscission scars, are useful for infrageneric delimitation. Variation in some anatomical characters, such as number of cell layers of collenchyma and cortex, number of cell layers of palisade parenchyma, size of petiole, number of lateral vascular bundles, exine ornamentation type of pollen, presence of large lumina in the middle of the primary lumen, the muri thickness, mericarp shape, mericarp length/width ratio and surface ornamentation type, can be used for separating species within the sections. Based on the updated geographical and ecological distribution, conservation status of the taxa is reassessed at regional, national and global levels. At the regional scale, the distribution is CR for 6 taxa EN for 9 taxa, VU for 5 taxa, NT for 10 taxa and LC for 29 taxa. At the national scale, the distribution is CR for 5 taxa, EN for 5 taxa, VU for 7 taxa, NT for 10 taxa and LC for 32 taxa. At the global scale, the distribution is CR for 5 taxa, EN for 3 taxa, VU for 6 taxa, NT for 10 taxa NT and LC for 35 taxa. The main threats in the study area are overgrazing, constructions, land clearing, fire, urbanization and tourism. The infrageneric delimitation is performed using multivariate analysis. Identification keys to sections and species are given. Synonymy, updated descriptions, phenology, distribution and habitats in Turkey, general distribution outside Turkey, distribution maps, phytogeography, specimen citations, some notes on taxonomy and photographs showing general appearance of the taxa are also provided.

The characteristic staminal lever mechanism is defined as a functional unit including the modification of stamens into lever-like structures, their reversible movement and the organisation with the remaining floral structures and they are involved in the process of pollen transfer. The staminal levers play a major role in the process of pollen deposition on the pollinators&rsquo / body and their proportion and interaction with pollinators may have significant consequences for the pollination system in Salvia. In this study, staminal lever mechanism and floral diversity of bee pollinated species including Salvia absconditiflora, Salvia aethiopis, Salvia russellii, Salvia tchihatcheffii which grow sympatrically are investigated in the campus flora of Middle East Technical University. The functioning of the staminal lever mechanism is tested and pollinators of the Salvia species are observed by field investigations. Special attention is given to species-specific patterns of pollen deposition on the pollinators&rsquo / body. Floral diversity of the Salvia species such as corolla shape, size, colour, stamen type are studied. Floral and pollinator structures are measured morphometrically by laboratory investigations. Moreover, the isolation mechanism among the co-occuring Salvia species are investigated by field observations. All field observations are backed by photos and video recordings. Field and laboratory investigations revealed that co-occuring Salvia species are reproductively isolated by several means and the diversity of floral constructions and staminal lever mechanism increases the mechanical isolation by attracting different pollinators and by loading pollen on different parts of their bodies.