The remarkable diversity of plant-feeding insects could be explained by the dynamics of their plant associations, where host plant shifts and specialization onto a small fraction of available plants may promote diversification. Neotropical Adelpha butterflies contain a large number of species, and previous work indicated the colonization of a novel host plant family (Rubiaceae) fueled its rapid diversification. However, accumulating host records indicate wide taxonomic host breadth at family level and below. Here, we categorize Adelpha diet breadth based on known host plant relationships across the Neotropics and from Costa Rica, Ecuador and Brazil. We also use a diet breadth index that identifies plants used in similar ways by Adelpha, pointing to potential plant traits that could facilitate or prevent plant-insect interactions. We find that diet breadth in Adelpha is not likely to change at different geographic scales, and that regional resource specialization was uncommon. Additionally, the diversification fueled by the switch to Rubiacae, appears to have led to some lowland-clade Adelpha species specializing on a restricted subset of host genera and species within Rubiaceae, as well as in Urticaceae. In contrast, the A. serpa-group shows generalization, with each species tending to feed on its own set of several unrelated plant families. Taken together, these results indicate that Rubiaceae and additional plant families appear as important ecological factors that have promoted adaptations in Adelpha and host plant family-level switches have not always had the same effect on diversification, corroborating the importance of Rubiaceae for this butterfly genus. Further research involving detailed phylogenies is needed to investigate associations between changes in diet breadth and speciation events, and test hypotheses of diet evolution.

Most of the species of the family Rubiaceae with flowers arranged in head inflorescences are currently classified in three distantly related tribes, Naucleeae (subfamily Cinchonoideae) and Morindeae and Schradereae (subfamily Rubioideae). Within Morindeae the type genus Morinda is traditionally and currently circumscribed based on its head inflorescences and syncarpous fruits (syncarps). These characters are also present in some members of its allied genera, raising doubts about the monophyly of Morinda. We perform Bayesian phylogenetic analyses using combined nrETS/nrITS/trnT-F data for 67 Morindeae taxa and five outgroups from the closely related tribes Mitchelleae and Gaertnereae to rigorously test the monophyly of Morinda as currently delimited and assess the phylogenetic value of head inflorescences and syncarps in Morinda and Morindeae and to evaluate generic relationships and limits in Morindeae. Our analyses demonstrate that head inflorescences and syncarps in Morinda and Morindeae are evolutionarily labile. Morinda is highly paraphyletic, unless the genera Coelospermum, Gynochthodes, Pogonolobus, and Sarcopygme are also included. Morindeae comprises four well-supported and morphologically distinct major lineages: Appunia clade, Morinda clade (including Sarcopygme and the lectotype M. royoc), Coelospermum clade (containing Pogonolobus and Morinda reticulata), and Gynochthodes-Morinda clade. Four possible alternatives for revising generic boundaries are presented to establish monophyletic units. We favor the recognition of the four major lineages of Morindeae as separate genera, because this classification reflects the occurrence of a considerable morphological diversity in the tribe and the phylogenetic and taxonomic distinctness of its newly delimited genera.

The identity of the long-overlooked Ronabea morindoides and Patabea tenuiflora, synonymous with a species of Appunia (Rubiaceae). Candollea 76: 83-92. In English, English abstract. The identity of Ronabea morindoides A. Rich. has long been unclear and is here investigated. Two sheets of original material corresponding to this name are deposited in the General Herbarium of the National Museum of Natural History of Paris (P), and represent a mixed collection; one part of this material corresponds better with the description of this taxon and is more unambiguously identifiable, and is here designated the lectotype. With this typification, Ronabea morindoides represents a species of Appunia Hook. f. The identity of Patabea tenuiflora DC. has also remained uncertain since its description and is here clarified by studying the holotype in the Candolle Herbarium (G-DC); this is an additional synonym of R. morindoides. Taxonomic review of this group in the Guianas also finds that Ronabea morindoides is an older name for Appunia brachycalyx (Bremek.) Steyerm. and Appunia surinamensis Bremek. (Morindeae). Therefore, the new combination Appunia morindoides (A. Rich.) Delprete, C.M. Taylor & T. McDowell is here published. Received: August 15, 2020; Accepted: December 1, 2020; First published online: February 1, 2021

L’étude chimique de l’exsudat glutineux recouvrant les bourgeons et la base des feuilles de Gardenia urvillei Montrouz. et Gardenia oudiepe Vieill., Rubiaceae endémiques de Nouvelle-Calédonie, a conduit à la détermination structurale de douze cycloartanes, neuf seco-cycloartanes, huit dammaranes et sept flavonoïdes. Parmi ces métabolites secondaires, six cycloartanes, quatre seco-cycloartanes et deux dammaranes sont des composés originaux. Les évaluations biologiques sur différentes cibles, notamment la recherche de propriétés anti-angiogéniques par déplacement du VEGF de son récepteur VEGF-R1 ou par inhibition de la polymérisation de la tubuline, ayant montré des résultats encourageants, une chimiothèque d’analogues a été hémisynthétisée. A cette fin, les composés majoritaires en séries cycloartane, dammarane, et flavonoïde, respectivement l’oudiépone A, l’hydroxydammarénone II, la santine, la 5,7-dihydroxy-3,3’,4’,5’,6-pentaméthoxyflavone et le kaempférol ont servi de points de départ à des modifications structurales ayant permis d’accéder à douze cycloartanes, deux seco-cycloartanes, trois dammaranes, un seco-dammarane et huit flavonoïdes supplémentaires. Un début de relation structure-activité a ainsi pu être décrit dans les domaines mentionnés précédemment. / Phytochemical studies of glutinous exudate covering the buds et the leaf base of Gardenia urvillei Montrouz. et Gardenia oudiepe Vieill., Rubiaceaeous species endemic to New Caledonia, have led to the identification of twelve cycloartanes, nine seco-cycloartanes, eight dammaranes et seven flavonoids. Among these secondary metabolites, six cycloartanes, four seco-cycloartanes et two dammaranes are original natural products. Biological evaluations, such as search for anti-angiogenic properties or inhibition of tubulin polymerization, have shown promising results. In order to establish the structure-activity relationships for these activities, twenty-six analogs were semisynthesized from the major isolated compounds, oudiépone A, hydroxydammarénone II, santin, 5,7-dihydroxy-3,3’,4’,5’,6-pentamethoxyflavone et kaempferol.

銀屑病是一種免疫相關的慢性炎症性皮膚病，其發病率約占世界人口的1-3%，而現今仍然缺乏有效安全的根治方法。國內外使用中草藥治療銀屑病取得較好的療效，但目前缺少對其進行系統研究和開發。我們研究小組之前對61種常用治療銀屑病中藥進行篩選, 發現中藥茜草根和南蛇藤的乙醇提取物具有強大的抑制表皮細胞增生的作用，本博士研究課題的目的是確定新的安全有效的用于治療銀屑病的中藥化學成分, 並闡明其作用機制。 / 本研究篩選了28種存在于這兩種中藥中的化學單體成分，采用體外培養永生化的人類皮膚良性角質形成細胞株HaCaT, 應用MTT法, 繪制細胞生長曲線，獲得抑制50%細胞生長所需藥物濃度(IC50)。實驗結果發現1-羟基-3-甲基蒽醌（HMA）， 1，4-二氨基-2，3-（2-苯氧基乙氧基）蒽醌 （DBA）和南蛇藤表現了強大的抗表細胞生長作用，其48小時培養後的IC50分別爲17.9，15.8，1.1 μM. 值得一提的是這些化合物對正常人表皮角質細胞HEK和人類成纖維細胞Hs68只有相對輕微細胞毒性。 / 隨後進行的機理研究，通過熒光染色，DNA凝膠電泳，細胞周期檢測，流式細胞計檢測及Western blot 分析結果表明， HMA和南蛇藤素是通過誘導細胞凋亡作用抑制HaCaT細胞生長。其中南蛇藤素通過線粒體凋亡和死亡受體介導的兩種通路誘導細胞凋亡， 其誘導細胞凋亡作用與其抑制核因子-κB在HaCaT細胞中的表達和活化有關。 / 另一方面，DBA 抑制人體表皮角質細胞生長的作用機理在于其對角質細胞終末分化的誘導作用。DBA與HaCaT和HEK細胞共同培養96小時後，能顯著促進細胞角質化外膜形成，同時上調角蛋白K1/10，人體套膜蛋白，轉谷氨酰胺酶-1表達和下調角蛋白K5/14表達。而利用小鼠尾部鱗片表皮模型對HMA的外用制劑進行測試，結果顯示HMA誘導角質細胞終末分化能力較弱。 / 總而言之，本研究課題從兩種中藥中成功發現三個具有較強的抗銀屑病活性的化學單體成分，這些來自中藥的天然産物具有很好的開發成新的銀屑病治療外用制劑的應用前景。 / Psoriasis is an immunologically-mediated chronic inflammatory disease of the skin and joints affecting approximately 1-3% of the world’s population. Traditionally, Chinese medicine has been extensively used both inside and outside China for treating psoriasis with promising clinical results. Based on the promising findings in our previous screening project on 61 psoriasis-treating Chinese medicines which showed the root of Rubia cordifolia L. (Rubiae Radix) to have potent anti-psoriatic action, the present study aimed to identify active anti-psoriatic chemical constituents derived from Rubiae Radix and another Chinese herb namely Celastrus orbiculatus Thunb. and to elucidate the underlying mechanisms of action. / Microplate MTT assay was performed to evaluate the anti-proliferative actions of 28 selected Rubiae Radix-derived anthraquinones and other chemical ingredients on cultured HaCaT keratinocytes. Among them, 1-hydroxy-3-methyl-anthraquinone (HMA) and 1,4-diamino-2,3-bis(2-phenoxyethoxy)anthraquinone (DBA), as well as celastrol, a Celastrus orbiculatus-derived triterpene, were found to possess significant anti-proliferative action on HaCaT cells, with IC₅₀ value of 17.9, 15.8 and 1.1 μM, respectively. All DBA, HMA and celastrol showed only mild to moderate toxic effects on normal human keratinocyte HEK cells and human fibroblast Hs68 cells. / Mechanistically, celastrol and HMA was found to induce apoptosis in a dose-dependent manner in HaCaT cells as characterized by DNA fragmentation, phosphatidyl-serine externalization and activation of caspase 3. Further studies by flow cytometric and western blot analyses demonstrated that the celastrol-induced apoptosis on HaCaT cells was associated with the inhibition of NF-κB pathway and through caspase-related apoptotic pathway as characterized by activation of caspase proteins, regulation of Bcl-2 family proteins and depolarization of mitochondrial potential. / On the other hand, DBA showed an ability to induce terminal differentiation in cultured human keratinocytes and this capability is believed to be responsible for its growth inhibitory effects. DBA significantly accentuated the cornified envelope formation in HEK and HaCaT keratinocytes together with the augmentation of K1/K10, involucrin and transglutaminase 1 protein levels and decrease of expression of K5/K14 protein in DBA-treated cells. However, the subsequent in vivo study using a mouse tail model showed that HMA did not have significant effects on modulating keratinocyte terminal differentiation. / Taken together, our present PhD project successfully identified DBA, HMA and celastrol to have potent anti-psoriatic action on in vitro models, and the experimental findings render these naturally-occurring chemicals to be promising candidates for further development into anti-psoriatic pharmaceutical agents. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Zhou, Linli. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2012. / Includes bibliographical references (leaves 213-244). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese. / Abstract (English) --- p.i / Abstract (Chinese) --- p.iii / Publications --- p.v / Acknowledgements --- p.vii / Table of Contents --- p.viii / List of Figures --- p.xvii / List of Tables --- p.xxi / List of Abbreviations --- p.xxii / Chapter Chapter One --- General Introduction / Chapter 1.1 --- Psoriasis --- p.2 / Chapter 1.1.1 --- Structure of skin --- p.2 / Chapter 1.1.2 --- Epidemiology of psoriasis --- p.3 / Chapter 1.1.3 --- Pathogenesis --- p.5 / Chapter 1.1.4 --- Classification --- p.8 / Chapter 1.1.4.1 --- Nonpustular (plaque type) psoriasis --- p.9 / Chapter 1.1.4.2 --- Guttate psoriasis --- p.9 / Chapter 1.1.4.3 --- Pustular psoriasis --- p.9 / Chapter 1.1.4.4 --- Erythrodermic psoriasis --- p.10 / Chapter 1.1.4.5 --- Nail psoriasis --- p.11 / Chapter 1.1.4.6 --- Psoriatic arthritis --- p.11 / Chapter 1.1.5 --- Comorbidities --- p.13 / Chapter 1.2 --- Treatment of Psoriasis --- p.16 / Chapter 1.2.1 --- Conventional treatment for psoriasis --- p.16 / Chapter 1.2.1.1 --- Topical therapy --- p.16 / Chapter 1.2.1.2 --- Phototherapy --- p.19 / Chapter 1.2.1.3 --- Systemic therapy --- p.21 / Chapter 1.2.2 --- Targeted immunotherapy --- p.24 / Chapter 1.2.3 --- Combination, rotational and sequential therapy --- p.25 / Chapter 1.2.4 --- Complementary treatment --- p.26 / Chapter 1.3 --- Traditional Chinese Medicine for Psoriasis --- p.30 / Chapter 1.3.1 --- Prescriptions for psoriasis based on pattern differentiation --- p.30 / Chapter 1.3.2 --- Clinical and experimental study of TCM for psoriasis --- p.34 / Chapter 1.3.3 --- Possible action mechanisms of Chinese herbs for psoriasis --- p.34 / Chapter 1.3.4 --- Previous studies on TCM for psoriasis conducted by our research group --- p.35 / Chapter 1.4 --- Aims and Objectives of the Present Study --- p.38 / Chapter Chapter Two --- Phytochemical and Apoptotic Studies of Rubiae Radix-derived Anthraquinones and Other Related Compounds / Chapter 2.1 --- Introduction --- p.41 / Chapter 2.2 --- Selection and Screening of Rubiae Radix-derived anthraquinones and Other Related Compounds for Anti-proliferative Action on Cultured HaCaT Human Keratinocytes --- p.43 / Chapter 2.2.1 --- Introduction --- p.43 / Chapter 2.2.2 --- Materials and methods --- p.45 / Chapter 2.2.2.1 --- Procurement of Rubiae Radix-derived anthraquiones and other related compounds --- p.45 / Chapter 2.2.2.2 --- Purification of anthraquinones from Rubiae Radix --- p.50 / Chapter 2.2.2.3 --- General cell culture --- p.54 / Chapter 2.2.2.4 --- SRB assay --- p.55 / Chapter 2.2.2.5 --- MTT assay --- p.56 / Chapter 2.2.2.6 --- Assessment of synergistic or antagonistic effects between two active anthraquiones --- p.56 / Chapter 2.2.2.7 --- Statistical analysis --- p.57 / Chapter 2.2.3 --- Results --- p.57 / Chapter 2.2.3.1 --- Anti-proliferative effects of 35 Rubiae Radix fractions on HaCaT cells by SRB assay --- p.57 / Chapter 2.2.3.2 --- Anti-proliferative effects of the 27 anthraquinones and related compounds on HaCaT cells by SRB assay --- p.59 / Chapter 2.2.3.3 --- Confirmation of the anti-proliferative action of 8 active pure compounds using MTT assay --- p.61 / Chapter 2.2.3.4 --- Cytotoxic effects of 1-hydroxy-3-methyl-anthraquinone and1,4-diamino-2,3-bis(2-phenoxyethoxy)anthraquinone on the growth of HEK and Hs68 cells --- p.64 / Chapter 2.2.3.5 --- Drug interactions between different active anthraquinones --- p.67 / Chapter 2.2.4 --- Discussion --- p.69 / Chapter 2.3 --- Investigations of the Apoptotic Effects of DBA and HMA on HaCaT cells --- p.71 / Chapter 2.3.1 --- Introduction --- p.71 / Chapter 2.3.2 --- Materials and methods --- p.76 / Chapter 2.3.2.1 --- Chemicals --- p.76 / Chapter 2.3.2.2 --- General cell culture methods --- p.76 / Chapter 2.3.2.3 --- Cell cycle analysis with PI staining --- p.76 / Chapter 2.3.2.4 --- Hoechst fluorescence staining for morphological evaluation --- p.77 / Chapter 2.3.2.5 --- DNA fragmentation assay --- p.77 / Chapter 2.3.2.6 --- Detection of apoptosis by flow cytometry --- p.78 / Chapter 2.3.2.7 --- Prepare cytosol fraction of HaCaT cells --- p.79 / Chapter 2.3.2.8 --- Western blot analysis --- p.79 / Chapter 2.3.2.9 --- Statistical analysis --- p.80 / Chapter 2.3.3 --- Results --- p.76 / Chapter 2.3.3.1 --- Action of DBA and HMA on cell cycle progression --- p.80 / Chapter 2.3.3.2 --- Alteration of cellular morphology --- p.84 / Chapter 2.3.3.3 --- Detection of DNA fragmentation --- p.86 / Chapter 2.3.3.4 --- Quantitative analysis of apoptotic cells by annexin V-PI staining --- p.88 / Chapter 2.3.3.5 --- Activation of procaspase-3 and release of cytochrome c protein --- p.91 / Chapter 2.3.4 --- Discussion --- p.94 / Chapter 2.4 --- General Discussion --- p.97 / Chapter Chapter Three --- Effects of Rubiae Radix and Its-derived Anthraquinones on Keratinocyte Terminal Differentiation / Chapter 3.1 --- Introduction --- p.100 / Chapter 3.2 --- Materials and Methods --- p.105 / Chapter 3.2.1 --- Chemicals --- p.105 / Chapter 3.2.2 --- General cell culture --- p.105 / Chapter 3.2.3 --- Cornified envelope (CE) formation assay --- p.106 / Chapter 3.2.4 --- Western blot analysis --- p.107 / Chapter 3.2.4 --- Statistical analysis --- p.107 / Chapter 3.3 --- Results --- p.108 / Chapter 3.3.1 --- EA fraction of Rubiae Radix, DBA and HMA stimulates CE formation --- p.108 / Chapter 3.3.2 --- EA fraction of Rubiae Radix, DBA and HMA regulated TG1 expression and involucrin production in cultured human keratinocytes --- p.112 / Chapter 3.3.3 --- Regulation of cytokeratins by EA fraction of Rubiae Radix, DBA and HMA --- p.118 / Chapter 3.4 --- Discussion --- p.128 / Chapter Chapter Four --- Anti-psoriatic Action of Celastrol from Celastrus orbiculatus / Chapter 4.1 --- Introduction --- p.136 / Chapter 4.2 --- Anti-proliferative Action of Celastrol on Cultured Human Keratinocytes and Other Cell Types --- p.138 / Chapter 4.2.1 --- Introduction --- p.138 / Chapter 4.2.2 --- Materials and methods / Chapter 4.2.2.1 --- Chemicals --- p.138 / Chapter 4.2.2.2 --- General cell culture --- p.139 / Chapter 4.2.2.3 --- MTT assay --- p.139 / Chapter 4.2.2.4 --- Statistical analysis --- p.139 / Chapter 4.2.3 --- Results --- p.142 / Chapter 4.2.3.1 --- Anti-proliferative effect of celastrol on cultured cells --- p.142 / Chapter 4.2.4 --- Discussion --- p.145 / Chapter 4.3 --- Induction of Apoptosis by Celastrol on Human Keratinocytes --- p.146 / Chapter 4.3.1 --- Introduction --- p.146 / Chapter 4.3.2 --- Materials and methods --- p.146 / Chapter 4.3.2.1 --- Chemicals --- p.146 / Chapter 4.3.2.2 --- General cell culture --- p.147 / Chapter 4.3.2.3 --- Cell cycle analysis with PI staining --- p.147 / Chapter 4.3.2.4 --- Detection of apoptosis by flow cytometry --- p.147 / Chapter 4.3.2.5 --- Measurement of the mitochondrial membrane potential (ΔΨm) --- p.148 / Chapter 4.3.2.6 --- Western blot analysis --- p.148 / Chapter 4.3.2.7 --- Statistical analysis --- p.148 / Chapter 4.3.3 --- Results --- p.149 / Chapter 4.3.3.1 --- Induction of sub-G1 phase by celastrol on HaCaT cells --- p.149 / Chapter 4.3.3.2 --- Quantitative analysis of apoptotic cells by Annexin V-PI staining --- p.151 / Chapter 4.3.3.3 --- Alteration of ΔΨm --- p.153 / Chapter 4.3.3.4 --- Activation of caspase family protein --- p.155 / Chapter 4.3.3.5 --- Celastrol regulates the Bcl-2 family members --- p.159 / Chapter 4.3.4 --- Discussion --- p.161 / Chapter 4.4 --- Inhibition of NF-κB Transcription Factor Activation by Celastrol --- p.164 / Chapter 4.4.1 --- Introduction --- p.164 / Chapter 4.4.2 --- Materials and methods --- p.165 / Chapter 4.4.2.1 --- Chemicals --- p.165 / Chapter 4.4.2.2 --- General cell cultrue --- p.165 / Chapter 4.4.2.3 --- Western blot analysis --- p.165 / Chapter 4.4.2.4 --- Detect nuclear p65 by ELISA assay --- p.166 / Chapter 4.4.2.5 --- Statistical analysis --- p.166 / Chapter 4.4.3 --- Results --- p.167 / Chapter 4.4.3.1 --- Celastrol inhibited the NF-κB activation --- p.167 / Chapter 4.4.4 --- Discussion --- p.170 / Chapter 4.5 --- Induction of Terminal Differentiation by Celastrol --- p.173 / Chapter 4.5.1 --- Introduction --- p.173 / Chapter 4.5.2 --- Materials and methods --- p.174 / Chapter 4.5.2.1 --- Chemicals --- p.174 / Chapter 4.5.2.2 --- General cell culture --- p.174 / Chapter 4.5.2.3 --- CE formation assay --- p.174 / Chapter 4.5.2.4 --- Western blot analysis --- p.174 / Chapter 4.5.2.5 --- Statistical analysis --- p.174 / Chapter 4.5.3 --- Results --- p.175 / Chapter 4.5.3.1 --- Regulation of CE formation by celastrol --- p.175 / Chapter 4.5.3.2 --- Modulation of terminal differentiation markers by celastrol --- p.178 / Chapter 4.5.4 --- Discussion --- p.181 / Chapter 4.6 --- General Discussion --- p.183 / Chapter Chapter Five --- In vivo Anti-psoriatic Effects of Topical Preparation of 1-hydroxy-3-methyl-anthraquinone / Chapter 5.1 --- Introduction --- p.187 / Chapter 5.2 --- Material and Methods --- p.191 / Chapter 5.2.1 --- Chemicals --- p.191 / Chapter 5.2.2 --- Formulation of topical preparation containing HMA --- p.191 / Chapter 5.2.3 --- Mouse tail model --- p.192 / Chapter 5.2.4 --- Histopathological evaluation --- p.193 / Chapter 5.2.5 --- Statistical analysis --- p.194 / Chapter 5.3 --- Results --- p.195 / Chapter 5.3.1 --- Body weight profile --- p.195 / Chapter 5.3.2 --- Histological resutls --- p.197 / Chapter 5.4 --- Discussion --- p.201 / Chapter Chapter Six --- General Conclusions and Future Perspectives / Chapter 6.1 --- General Conclusions --- p.205 / Chapter 6.2 --- Future Perspectives --- p.210 / References / References by alphabetical order --- p.213

銀屑病是一種慢性炎症性皮膚病，其發病率約佔全球1-3%的人口。銀屑病的病理特徵包括角質細胞增殖和分化異常，同時伴隨炎症反應，白細胞聚集於真皮和表皮以及血管擴張。證據顯示角質細胞能參與及延續免疫反應，以達致維持或促進該病的作用。研究亦建議角質細胞減少凋亡是引致銀屑病的一個特定現象；因此，長期以來誘導角質細胞凋亡就被用作為治療銀屑病的一種有效策略。 / 根據銀屑病的嚴重程度，治療方法可分為三級：外用藥物主要用於比較輕微的病患，而光療適合中等程度的病患；對於嚴重病例則可使用系統性治療或生物製劑。基於大約75%的銀屑病患者屬於輕微至中度病患，外用藥物是目前應用最為廣泛的治療方法。在中國銀屑病治療的歷史中曾經使用過中草藥，研究亦表明，其治療機制可能通過抑制角質細胞增殖和誘導角質細胞凋亡。比較研究也指出，傳統中藥比西藥的副作用相對較少，及具有較長的舒緩期和較低的復發率。 / 我們先前的研究發現，茜草根提取物能夠抑制一個和銀屑病相關的HaCaT角質細胞增殖。本研究證實，茜草根的乙酸乙酯提取物（EA）能誘導HaCaT細胞凋亡，其抑制角質細胞增殖的作用比茜草根的乙醇提取物（EE）更為有效，並可和一個流行於歐洲國家的重要外用銀屑病治療藥地蒽酚相比。另外，透過不同的檢測，包括形態學觀察，細胞凋亡雙染（磷脂結合蛋白V-碘化丙啶）分析，細胞週期分析，去氧核醣核酸斷裂測試，原位末端轉移酶標記技術，免疫熒光染色以及西方墨點法，我們發現一種在茜草中的化合物，1,4-二羥基-2-萘甲酸（DHNA）能通過死亡受體介導，線粒體介導或不依賴胱天蛋白酶的途徑導致HaCaT細胞凋亡。同時，在其中一種銀屑病動物模型，小鼠鼠尾鱗片表皮上的初步研究顯示DHNA亦可誘導角質細胞分化。此外，在細胞水平（存活率，釋放白细胞介素-1α）和動物上（Draize動物皮膚刺激性試驗）的實驗結果表明DHNA比地蒽酚的刺激性較小。 / 總括而言，本研究透過人類皮膚細胞和動物實驗說明EA和DHNA的細胞凋亡機制，以及DHNA對皮膚的潛在刺激性。這些結果顯示EA和DHNA有潛能發展成為安全及能有效治療銀屑病的替代藥物。EA和DHNA可在一個連續療程中結合使用，其中EA藥效媲美地蒽酚，應能迅速清除銀屑病皮損；而DHNA比地蒽酚的刺激性小，則比較適合應用在這個連續療程中後來的維護保養階段 / Psoriasis is a chronic inflammatory skin disorder that affects approximately 1-3% of the population worldwide. It is characterized by epidermal hyperplasia or abnormal differentiation, infiltration of leucocytes into the dermis and epidermis, dilation of blood vessels in dermis and inflammation. Evidence indicates keratinocytes contributed to the disease, and keratinocytes also participate in maintaining the chronically perpetuating immune response that sustains psoriasis. Decrease in keratinocytes apoptosis is suggested to be a specific pathogenic phenomenon, and induction of keratinocytes apoptosis have long been considered as an effective anti-psoriatic strategy. / Treatment of psoriasis is based on disease severity. Topical agents are predominantly for mild conditions; phototherapy for moderate conditions and systemic treatment or biological agents for severe cases. Topical treatment remains the most widely used method as an estimated 75% of psoriatic patients have mild to moderate disease. Chinese herbs have been used for the treatment of psoriasis in China, and studies showed their mechanism on treating psoriasis may through inhibition of keratinocyte proliferation and induction of apoptosis. Comparison studies also show that traditional Chinese medicine has relatively fewer side effects than western therapeutic agents, with a longer remission time and lower recurrence rate. / The extract of the root of Rubia cordifolia L. (Rubiae Radix et Rhizoma) was previously found to inhibit keratinocyte proliferation using a psoriasis-relevant HaCaT cells model. In this study, the ethyl acetate extract of the root of Rubia cordifolia L. (EA) was confirmed to induce apoptosis on HaCaT cell, and the antiproliferative effect of EA is more potent than the ethanol extract of the herb (EE) and is comparable to dithranol, an important and popular topical treatment for psoriasis among Europe countries. Besides, we identified one of the components in Rubia cordifolia L., 1,4-dihydroxy-2-naphthoic acid (DHNA), could induce HaCaT keratinocyte apoptosis through the death receptor and mitochondria mediated pathway as well as in a caspase independent manner using various assays such as morphological examination, annexin V-PI staining, cell cycle analysis, DNA fragmentation, TUNEL assay, immunofluorescence staining and Western blot analysis. Moreover, DHNA was found to induce keratinocyte differentiation in a preliminary study using the in vivo mouse tail model of psoriasis. Furthermore, results from in vitro (cell viability, IL-1α release) and in vivo (Draize animal skin irritation test) experiments suggested DHNA have less irritation problems than dithranol. / In summary, this study describes the apoptotic mechanism of EA and DHNA, as well as the irritation potential of DHNA using different human skin cells and animal model. These results suggest EA and DHNA have the potential to develop as safe and effective therapeutic alternative for the treatment of psoriasis. EA and DHNA can be used together in a sequential therapy, in which EA is effective in rapid clearing of psoriatic lesions as its potency is comparable to dithranol; whereas DHNA is better suited for the later maintenance therapy for its milder irritation effect compared with dithranol. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Mok, Chong Fai. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2012. / Includes bibliographical references (leaves 164-183). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese. / Abstract (English version) --- p.iv / Abstract (Chinese version) --- p.vi / List of Publication and Presentation --- p.viii / Acknowledgements --- p.ix / Table of Contents --- p.x / List of Tables --- p.xvi / List of Figures --- p.xvii / List of Abbreviations --- p.xx / Chapter Chapter 1: --- Introduction --- p.1 / Chapter 1.1. --- Psoriasis --- p.1 / Chapter 1.1.1. --- Histological features --- p.2 / Chapter 1.1.2. --- Role of keratinocytes in psoriasis --- p.4 / Chapter 1.1.3. --- Decrease in skin cell apoptosis --- p.8 / Chapter 1.2. --- Treatment of psoriasis --- p.9 / Chapter 1.2.1. --- Conventional treatment --- p.9 / Chapter 1.2.1.1. --- Mild disease --- p.10 / Chapter 1.2.1.1.1. --- Corticosteroids --- p.10 / Chapter 1.2.1.1.2. --- Vitamin D₃ analogs --- p.11 / Chapter 1.2.1.1.3. --- Tazarotene --- p.11 / Chapter 1.2.1.1.4. --- Anthralin --- p.12 / Chapter 1.2.1.1.5. --- Coal tar --- p.12 / Chapter 1.2.1.2. --- Moderate disease --- p.12 / Chapter 1.2.1.2.1. --- Phototherapy --- p.12 / Chapter 1.2.1.3. --- Severe disease --- p.13 / Chapter 1.2.1.3.1. --- Retinoids --- p.13 / Chapter 1.2.1.3.2. --- Methotrexate --- p.14 / Chapter 1.2.1.3.3. --- Cyclosporine --- p.14 / Chapter 1.2.1.3.4. --- Fumaric acid --- p.15 / Chapter 1.2.1.3.5. --- Biological agents --- p.15 / Chapter 1.2.2. --- Alternative treatment --- p.16 / Chapter 1.2.2.1. --- Traditional Chinese Medicine (TCM) --- p.17 / Chapter 1.3. --- Aims and objectives of the present study --- p.19 / Chapter Chapter 2: --- Apoptotic Action of Ethyl Acetate Fraction of the Root of Rubia cordifolia L. (Rubiae Radix et Rhizoma) on HaCaT Human Keratinocytes --- p.21 / Chapter 2.1. --- Introduction --- p.21 / Chapter 2.1.1. --- Rubia cordifolia L. --- p.21 / Chapter 2.1.2. --- Apoptosis --- p.22 / Chapter 2.1.3. --- Study objectives --- p.28 / Chapter 2.2. --- Materials and Methods --- p.30 / Chapter 2.2.1. --- Sources of medicinal materials --- p.30 / Chapter 2.2.2. --- Preparation of extracts --- p.30 / Chapter 2.2.3. --- Reagents --- p.31 / Chapter 2.2.4. --- Cell culture --- p.31 / Chapter 2.2.5. --- Proliferation assay --- p.32 / Chapter 2.2.6. --- Fluorescent staining for morphological evaluation --- p.33 / Chapter 2.2.7. --- Annexin V/propidium iodide staining --- p.33 / Chapter 2.2.8. --- JC-1 staining --- p.34 / Chapter 2.2.9. --- Statistical analysis --- p.35 / Chapter 2.3. --- Results --- p.36 / Chapter 2.3.1. --- EA inhibits proliferation of human epidermal HaCaT keratinocytes --- p.36 / Chapter 2.3.2. --- Alteration of cellular morphology --- p.39 / Chapter 2.3.3. --- EA increases phosphatidylserine externalization in HaCaT cells --- p.41 / Chapter 2.3.4. --- EA decreases MMP --- p.45 / Chapter 2.4. --- Discussion --- p.47 / Chapter Chapter 3: --- Identification of Pure Compound for Possible Apoptotic Action on HaCaT Human Keratinocytes and Detailed Mechanistic Study --- p.51 / Chapter 3.1. --- Introduction --- p.51 / Chapter 3.1.1. --- Anthraquinone --- p.51 / Chapter 3.1.2. --- Study objectives --- p.52 / Chapter 3.2. --- Materials and Methods --- p.54 / Chapter 3.2.1. --- Reagents --- p.54 / Chapter 3.2.2. --- Cell culture --- p.54 / Chapter 3.2.3. --- Proliferation assay --- p.55 / Chapter 3.2.4. --- Fluorescent staining for morphological evaluation --- p.56 / Chapter 3.2.5. --- Annexin V/propidium iodide staining --- p.56 / Chapter 3.2.6. --- JC-1 staining --- p.56 / Chapter 3.2.7. --- Cell cycle analysis --- p.56 / Chapter 3.2.8. --- Detection of DNA fragmentation --- p.57 / Chapter 3.2.9. --- Terminal Deoxynucleotidyltransferase-Mediated dUTP Nick End Labeling (TUNEL) assay --- p.57 / Chapter 3.2.10. --- Western blot analysis --- p.58 / Chapter 3.2.11. --- Immunofluorescence staining --- p.59 / Chapter 3.2.12. --- Statistical analysis --- p.60 / Chapter 3.3. --- Results --- p.61 / Chapter 3.3.1. --- DHNA inhibits proliferation of human epidermal HaCaT Keratinocytes --- p.61 / Chapter 3.3.2. --- Alteration of cellular morphology --- p.70 / Chapter 3.3.3. --- DHNA increases phosphatidylserine externalization in HaCaT cells --- p.72 / Chapter 3.3.4. --- DHNA decreases MMP --- p.76 / Chapter 3.3.5. --- DHNA causes G0/G1 cell cycle arrest in HaCaT cells --- p.78 / Chapter 3.3.6. --- DHNA increases DNA fragmentation --- p.81 / Chapter 3.3.7. --- DHNA increases TUNEL positive cells in HaCaT cells --- p.83 / Chapter 3.3.8. --- Western blot analysis --- p.85 / Chapter 3.3.9. --- DHNA induced Fas aggregation in HaCaT cells --- p.88 / Chapter 3.3.10. --- Caspase inhibition assay --- p.90 / Chapter 3.3.11. --- DHNA induced caspase independent apoptosis in HaCaT cells --- p.93 / Chapter 3.3.12. --- Effects of DHNA on MAPK in HaCaT cells --- p.96 / Chapter 3.3.13. --- MAPK inhibition assay --- p.100 / Chapter 3.4. --- Discussion --- p.104 / Chapter Chapter 4: --- Anti-Psoriatic Effects of Topical 1,4-Dihydroxy-2-naphthoic acid Formulation on in vivo Mouse Tail Experiments --- p.111 / Chapter 4.1. --- Introduction --- p.111 / Chapter 4.1.1. --- Keratinocytes differentiation process --- p.111 / Chapter 4.1.2. --- Animal model for psoriasis --- p.114 / Chapter 4.1.3. --- Study objectives --- p.119 / Chapter 4.2. --- Materials and Methods --- p.122 / Chapter 4.2.1. --- Reagents --- p.122 / Chapter 4.2.2. --- Formulation and preparation of topical drug --- p.122 / Chapter 4.2.3. --- Mice for in vivo experiments --- p.123 / Chapter 4.2.4. --- Treatment with topical preparations --- p.124 / Chapter 4.2.5. --- Statistical analysis --- p.125 / Chapter 4.3. --- Results --- p.126 / Chapter 4.3.1. --- Tail skin appearance after topical treatment --- p.126 / Chapter 4.3.2. --- Histological examination and findings --- p.128 / Chapter 4.4. --- Discussion --- p.132 / Chapter Chapter 5: --- Prediction of Skin Irritation Potential of 1,4-Dihydroxy-2-naphthoic acid by in vitro and in vivo Experiments --- p.135 / Chapter 5.1. --- Introduction --- p.135 / Chapter 5.1.1. --- Skin irritation --- p.135 / Chapter 5.1.2. --- Viability test and IL-1α release --- p.136 / Chapter 5.1.3. --- Animal irritation test --- p.139 / Chapter 5.1.4. --- Study objectives --- p.139 / Chapter 5.2. --- Materials and Methods --- p.141 / Chapter 5.2.1. --- Reagents --- p.141 / Chapter 5.2.2. --- Cell culture --- p.141 / Chapter 5.2.3. --- Viability test --- p.141 / Chapter 5.2.4. --- IL-1α release assay --- p.142 / Chapter 5.2.5. --- Animal irritation test --- p.142 / Chapter 5.2.6. --- Statistical analysis --- p.143 / Chapter 5.3. --- Results --- p.144 / Chapter 5.3.1. --- Viability test --- p.144 / Chapter 5.3.2. --- IL-1α release assay --- p.144 / Chapter 5.3.3. --- Animal irritation test --- p.147 / Chapter 5.4. --- Discussion --- p.152 / Chapter Chapter 6: --- General Discussion and Conclusions --- p.155 / References --- p.164

Plant-based defence mutualisms involve aspects of plant morphology that influence the performance of plant parasites, their natural enemies and trophic interactions. Leaf domatia, small indentations on the underside of leaves, can be structurally complex, and are often inhabited by potentially beneficial mites and other arthropods. Plant morphological traits such as domatia that enhance mutualistic relationships may result in increased plant growth rates, and reproductive success. New Zealand supports ~60 plant species that have domatia, the most speciose genus being Coprosma. The aim of this thesis was to examine factors that affect the production of leaf domatia and their relationship with foliar mite assemblages. The three main objectives of this thesis are: First, to investigate the production of foliar domatia and their susceptibility to limited resources, particularly to carbon availability. Second, to test if domatia are inducible structures during leaf ontogeny in the presence of foliar mites and/or fungi. Finally, to explore the effect of domatia availability on foliar mite assemblages on leaves with and without resident mites. This thesis tested the stated objectives using C. lucida, C. ciliata, C. foetidissima and C. rotundifolia, with a combination of field investigations and controlled manipulative experiments. The cost of domatia production was investigated using two field surveys and two controlled experiments. Under natural conditions the relationship between leaf morphology and domatia were measured in situ and across an altitudinal gradient. The experimental manipulations used carbon and nutrient stress, induced by temperature, light and fertilizer application. The second objective was experimentally tested under field conditions by manipulating foliar mites and fungal densities on C. rotundifolia. The third objective was investigated by manipulating domatia availability on C. lucida shrubs across three different vegetation types. Under field conditions, the number of domatia per leaf was associated with leaf morphology in C. lucida and C. foetidissima, but not C. rotundifolia. Foliar carbon showed a positive, but weak association with domatia production in C. foetidissima and C. ciliata. Altitudinal induced-carbon stress on domatia production was ambiguous. Domatia production in C. foetidissima was positively associated to altitude in field survey (1), and negatively associated in the second survey, with no correlation found between carbon and altitude. Experimental C. rotundifolia shrubs held under elevated night-time temperatures showed a 2.5 fold increase in respiration, a 34% to 91% decrease in daily carbon gain, and 38% decrease in domatia per leaf mass. Domatia production showed no significant differences under nutrient stress. The results showed little evidence to support a role for induction of domatia. Domatia production in new leaves was similar across all experimental treatments. Diverse vegetation types supported 60% higher mite species. Leaves with domatia supported ~22 to 66% higher mite densities, greater colonisation success and more diverse mite assemblages, than those without domatia. In the pastoral vegetation, the absence of predatory mites on experimental shrubs resulted in no differences in fungivorous mite densities regardless of domatia availability. Plant investment in foliar domatia appears associated with the number of available sites on the leaf under field conditions. The role of carbon availability during leaf ontogeny suggests a complex and highly variable association with domatia production. Domatia are constitutive defence structures that influence mite assemblages, mediating both beneficial and antagonistic relationships. This thesis concludes that domatia are in part, carbon-based non-inducible structures that influence mite assemblages, plant-mite and mite-mite interactions, and increase the probability of successful colonisation.

The phylogenetic relationships within subfamily Ixoroideae of the coffee family are investigated by phylogenetic reconstruction of molecular data, including regions of the chloroplast DNA (matK, ndhF, rbcL, rps16, trnH-psbA, trnS-G, and trnT-F), and the nuclear ribosomal DNA (ITS). The evolution of morphological characters within the group are inferred, with focus on characters used in classification. Ixoroideae have primarily been characterized by secondary pollen presentation, contorted corolla aestivation, and fleshy fruits. Secondary pollen presentation appears synapomorhic of a clade comprising the Ixoroideae crown group together with Retiniphyllum, whereas contorted corolla aestivation has evolved earlier and is synapomorphic for the crown group, Retiniphyllum, and Steenisia. Capsules likely represent a plesiomorphy from which various dry or fleshy indehiscent fruits have evolved independently in different clades. Reductions in seed number have also occured in many clades, none of which shows a secondary increase in the number of seeds. Within Ixoroideae, the phylogeny and tribal delimitations of Alberteae and Condamineeae are studied in more detail. The former appears restricted to Alberta, Nematostylis, and Razafimandimbisonia, a new genus described here. The Condamineeae are a diverse tribe largely unresolved in previous molecular phylogenetic studies. Our results support a synonymization of both Calycophylleae and Hippotideae, because these are nested within the Condamineeae. Ancestral state reconstructions indicate that intrapetiolar stipules, poricidal anthers, and protogyny, otherwise uncommon characters in Rubiaceae, all have evolved more than once in the Condamineeae. The rare genera Jackiopsis, Glionnetia, and Trailliaedoxa previously not included in molecular phylogenetic analyses, are all found nested within the Ixoroideae, and their systematic positions are discussed. The genera Bathysa, Calycophyllum, Elaeagia, and Rustia do not appear monophyletic. Consequently, resurrections of the names Holtonia, Schizocalyx, and Semaphyllanthe, and synonymizations of Phitopis (as Schizocalyx) and Tresanthera (as Rustia) are proposed. Also proposed are five new tribal names for clades that are not associated with any previously described tribes in the phylogenetic hypotheses presented. / At the time of the doctoral defense, the following papers were unpublished and had a status as follows: Paper 3: Manuscript. Paper 4: Manuscript.

La Nouvelle-Calédonie est un archipel du Pacifique Sud-Ouest. Sa flore est riche, unique, et dysharmonique. L'origine temporelle et géographique de cette flore constitue une des problématiques majeures des botanistes. Une étude systématique a été menée sur le genre Psychotria (Rubiaceae), et ses genres alliés (clade Psychotrieae-Palicoureeae), afin de comprendre les modes d'établissement et de diversification des plantes dans l'archipel. Les deux tribus comportent 86 espèces néo-calédoniennes réparties entre les genres Geophila (une espèce), Margaritopsis (quatre espèces) et Psychotria (81 espèces). L'étude taxonomique a permis de déterminer pour chacun d'eux les noms d'espèces valides, et d'identifier 26 espèces nouvelles de Psychotria et Margaritopsis. Neuf combinaisons et noms spécifiques nouveaux sont proposés.La Nouvelle-Calédonie a été colonisée au moins quatre fois par les Psychotrieae-Palicoureeae. Bien que leurs arrivées soient simultanées durant le Néogène, ces quatre lignées ont des histoires évolutives différentes à mettre en lien avec leurs modes propres d'établissement et de diversification. Le genre Geophila n'a pas diversifié. Les Margaritopsis ont eu une diversification modeste, à l'image des autres espèces des îles du Pacifique. Les Psychotria clade NC1 constituent une lignée relique, ayant probablement subit une extinction, et qui se serait uniquement maintenue dans les maquis miniers. Les Psychotria cladeNC2 constituent la plus large, jeune et rapide radiation de plantes de la Nouvelle-Calédonie, probablement originaire des forêts humides d'Australie.Le clade NC2 est en pleine expansion évolutive. Il est constitué de 12 lignées internes. Sa capacité à tolérer les substrats ultramafiques, acquise avant son arrivée dans l'archipel, a favorisé son établissement local. Sa large diversité a probablement différentes origines : une labilité vis-à-vis de la nature des substrats géologiques, des changements éventuels de pollinisateurs, un changement de niche écologique amorcé, lié à l'acquisition d'adaptations à la sécheresse et à l'avènement d'un climat plus aride en Nouvelle-Calédonie durant le Pliocène.

Infections caused by microorganisms are a serious public health problem, mainly due to the emergence of multi drug resistant bacteria and fungi. Thus, the search for new alternative therapeutic approaches remains, being the goal of several research groups worldwide. The World Health Organization (WHO) recognizes the value and encourages the use of traditional medicine knowledge. Brazil has a large number of plant species and many of them are considered medicinal. However, despite quite a lot of them are popularly used they were not scientifically validated yet. The Northeast region hosts an extensive native flora with many species showing therapeutic properties, but most of them scientifically unproven. As an example, there were representatives of the families Rubiaceae and Polygonaceae, the latter being the fourth largest family between angiosperms. Amongst Alagoas flora there are species of these two families, such as Coccoloba mollis Casaretto (1844) (Polygonaceae), Triplaris americana Linnaeus (Polygonaceae) and Coutarea hexandra Schum. (Rubiaceae), which genera have species cited by its ethnomedicinal uses. The aim of this study was to evaluate the antimicrobial activity of crude extracts and fraction obtained from these plant species. Tests were carried out in Petri dishes by the disk diffusion method to evaluate relative effectiveness of tested samples as antimicrobial agents, by measuring zones of inhibition on culture of Staphylococcus aureus (ATCC 25923), Pseudomonas aeruginosa (ATCC 27853) e Candida albicans (ATCC 10231). The minimal inhibitory concentration (MIC) was determined by microdilution in BHI broth (Brain Heart Infusion). Plant species were also tested against Artemia salina larvae for preliminary evaluation of its acute toxicity. Of the 69 samples tested, 17 were effective against S. aureus. The methanol fraction from the dichloromethane extract of the roots of C. hexandra was considered the most promising because it presented the highest percentage bacterial growth inhibition. Of the samples subjected to MIC determination all were active with values ranging from 125 to 500 μg/mL. None of the tested species were active against P. aeruginosa or C. albicans. Regarding the toxicity on A. salina, in preliminary tests 32 of the 69 tested samples were non-toxic. The others were submitted to quantitative test 10 of them exhibited non-toxic effects. Among the samples that showed toxicity LC50 values ranged from 2.68 to 953.9 g/mL. The three plant species showed antibacterial activity against S. aureus. For toxicity against A. salina, the 27 toxic samples, 17 are species of the family Polygonaceae, and the rest of the species C. hexandra were toxic / Fundação de Amparo a Pesquisa do Estado de Alagoas / As infecções causadas por microrganismos são um sério problema de saúde pública, principalmente devido emergência de bactérias e fungos multiresistentes. Portanto, permanece a necessidade da busca de novas alternativas de tratamento, objetivo de vários grupos de pesquisa no mundo. A Organização Mundial de Saúde (OMS) reconhece o valor e incentiva o aproveitamento do conhecimento tradicional. O Brasil possui um grande acervo vegetal, e muitas plantas são consideradas medicinais. No entanto, apesar de várias delas serem utilizadas popularmente ainda não foram validadas cientificamente. A região Nordeste abriga uma vasta flora nativa com muitas espécies com propriedades terapêuticas, porém a maior parte ainda sem comprovação científica. Como exemplo, citam-se representantes das famílias Polygonaceae e Rubiaceae, sendo esta última a quarta família de maior diversidade entre as angiospermas. Na flora Alagoana encontram espécies destas duas famílias, como a Coccoloba mollis Casaretto (1844) (Polygonaceae), Triplaris americana Linnaeus (Polygonaceae) e Coutarea hexandra Schum. (Rubiaceae), cujos gêneros possuem espécies com registros sobre seu uso etnomedicinal. O objetivo deste trabalho foi avaliar a atividade antimicrobiana de extratos brutos e frações obtidas destas espécies vegetais. Os testes foram conduzidos em placas de Petri pelo método de difusão em disco para avaliar o potencial antimicrobiano das amostras testadas, pela formação de halo de inibição frente à cultivos de Staphylococcus aureus (ATCC 25923), Pseudomonas aeruginosa (ATCC 27853) e Candida albicans (ATCC 10231). A concentração inibitória mínima (CIM) foi determinada pela técnica de microdiluição em caldo BHI (infusão de cérebro e coração). As espécies vegetais foram também testadas para avaliação preliminar de sua toxicidade aguda, frente a larvas de Artemia salina. Das 69 amostras testadas, 17 delas apresentaram atividade contra S. aureus, a fração em metanol proveniente de extrato em diclorometano da raiz de C. hexandra foi considerada a mais promissora por ter apresentado maior percentual de inibição do crescimento bacteriano. Submetidas a determinação da CIM todas foram ativas com valores entre 125 e 500 μg/mL. Nenhuma das espécies vegetais apresentou atividade contra P. aeruginosa, ou contra C. albicans. Com relação à toxicidade frente A. salina, das 69 amostras submetidas ao teste preliminar 32 foram atóxicas. As demais foram submetidas ao teste quantitativo sendo 10 atóxicas. Entre as amostras que apresentaram toxicidade os valores de CL50 variaram de 2,68 a 953,9 g/mL. As três espécies vegetais apresentaram atividade antibacteriana frente S. aureus, com relação a toxicidade frente A. salina, das 27 amostras tóxicas, 17 são das espécies da família Polygonaceae, e as demais da espécie C. hexandra foram tóxicas.