Echinacea purpurea is consumed as a natural health product around the world. Due to the genus’ ethnobotanical relevance, the phytochemistry of Echinacea has been extensively studied, revealing a variety of bioactive metabolites including caffeic acid derivatives and alkylamides. Whereas seasonal trends in root chemistry have been established, trends in other plant parts are relatively understudied. Similarly, few studies have evaluated the effects of organic plant growth substances in field trials. With increased demand for organic products, industry is looking for alternative ways to optimize yields and medicinal properties. For this thesis, my first objective was to quantify the concentrations of E. purpurea’s secondary metabolites across organic treatments throughout the plant’s first growth year to determine optimal harvesting time and conditions in all parts of the plant. The second objective was to determine how seasonal variations affect its potential bioactivity through inhibition of Pseudomonas aeruginosa. Plants were grown in field plots treated with four different organic treatments: water (control), high cytokinin, low cytokinin, and fish oils; samples were collected biweekly from May-September. Dried plants were separated into major plant parts and were extracted exhaustively in 70% EtOH. Using high-pressure liquid chromatography (HPLC), concentrations of alkylamides and select caffeic acid derivatives were quantified in all samples and compared across plant part, developmental stage, and organic fertilizers. It was determined that while there were no major differences between treatments, phytochemical concentrations changed throughout the season in all plant parts; revealing that aerial parts of the plant also bioactive secondary metabolites and should not be excluded from future studies. Following this study, an MIC50 assay was used to test these extracts against P. aeruginosa PA14. It was noted that seasonality effects of phytochemistry were not consistent with bioactivity and that there were no significant differences between extract and carrier control. While the antibiotic activity of root extracts varied seasonally, the flower extract exhibited the most consistent antibiotic potential. The results presented in this thesis will not only aid in industry practices and yield optimization but, through filling knowledge gaps on seasonality and organic treatments in field trials, will increase the understanding of E. purpurea’s chemistry and related bioactivity, with implications on both the medicinal properties and eco-physiology of the species.

This thesis describes the in vivo evaluation of orally administered extracts of Echinacea species, and fractions obtained from Echinacea purpurea for specific immunostimulatory function induced in Balb/c mice by immunization with microbial vaccines. Two vaccines were used for this purpose in this study. The first vaccine used consisted of whole-cell formalin-killed Salmonella typhimurium vaccine, which is known to induce production of specific antibodies but no delayed-type hypersensitivity response considered as an indicator of the cell-mediated immune response. The second vaccine was a commercial acellular pertussis vaccine (DTaP) consisting of inactivated diphtheria and tetanus toxoids and native or inactivated virulence factors of Bordetella pertussis (the etiological agent of whooping cough) which is known to induce a humoral immune response but with controversial reports about performance in the induction of cell-mediated immunity. Both vaccines were administered intraperitoneally, whereas the herbal extracts and fractions were administered by oral gavage. The phytochemical profile of the ethanolic extracts of Echinacea purpurea, Echinacea pallida, Echinacea angustifolia and one commercial product "Echinacea Triplex" and three fractions prepared from Echinacea purpurea, namely polysaccharides, phenolics and alkylamides, was determined by thin-layer chromatography, high-performance liquid chromatography and/or by chemical analytical techniques. Statistically significant increases in specific anti-Salmonella typhimurium serum antibody were obtained for the Echinacea purpurea, Echinacea angustifolia and Echinacea Triplex [Trade Mark] treatment groups, and also for groups receiving treatment with fractions obtained from Echinacea purpurea, namely, polysaccharide, alkylamide and phenolic fractions. Statistically significant increases in the anti-B pertussis serum antibodies were also obtained in the young Echinacea Triplex [Trade Mark] group vaccinated with the DTaP vaccine. Significant enhancement of serum interleukin-12 titres was observed in the Echinacea Triplex [Trade Mark] as well as the phenolic fraction treatment groups which were vaccinated with killed Salmonella typhimurium vaccine. Of the groups vaccinated with the DTaP vaccine, only Echinacea Triplex [Trade Mark] displayed statistically significant increases in mean interleukin-12 titre compared to the placebo in both young and old groups, however Echinacea purpurea, and Echinacea angustifolia both displayed significant increases in mean interleukin-12 titre in the young treatment mice. Increases in interferon-gamma levels in mice orally dosed with phenolic, polysaccharide or alkylamide fractions of Echinacea purpurea and vaccinated with the killed Salmonella typhimurium vaccine were also observed, however these need to be repeated for confirmation, and statistical analysis.

M.Tech. (Homoeopathy) / The purpose of this study is to determine the effect of the homoeopathic complex formula consisting of , Echinacea angustifolia tincture, Baptisia tinctoria tincture and Thuja occidentalis D1 on circulating leukocytes. These medicinal plants have immune enhancing properties and an effect on leukocytes. Leukocytes play an important role in the body's defence system. 54 volunteers participated in a randomised double blind placebo controlled trial. An initial full blood count before treatment served as a base-line control for each volunteer. A total of 15ml of the medication placebo was administered over 3 hours. After the last dose, a further three blood samples were drawn at 90 minute intervals. Blood analysis included a differential leukocyte count. The trial was conducted over a single 6 hour period. The homoeopathic medication caused a decrease in circulating leukocytes, specifically lymphocytes. The efficacy of this medicationas an immunostimulant should be confirmed on a larger study sample.

Bakgrund Influensa är en akut respiratorisk infektion som orsakas av ett negativt RNA-virus. Insjuknade i influensa sker ofta under vintertid. Influensa finns i fyra olika typer, A-D. A är den vanligaste sorten under influensasäsong och påverkar både människor och djur, medan D påverkar främst grisar och nötkreatur. Influensa kännetecknas av att det börjar med en frossa och med en snabbt stigande feber (över 40℃) samt med muskelvärk och med en märkbar sjukdomskänsla. Efteråt sprids infektionen till luftvägarna, där det uppkommer symtom i form av torrhosta. Influensa sprids genom luftburna aerosoler eller genom dropp- och kontaktsmitta. Influensaviruset tar sig in i cellen via sina ytproteiner hemagglutinin (HA) och neuraminidas (NA) och sedan tar det sig in till värdcellens kärna, vilket leder till replikation av viruset. Vid influensasäsongen ges vaccinationer till personer som är över 65 år, och till är gravida och till personer som har någon underliggande kronisk sjukdom. Detta görs för att förhindra allvarlig sjukdom och för att kunna minska antalet komplikationer och dödsfall under influensasäsongen. Neuraminidashämmare som exempelvis oseltamivir och zanamivir kan ges för smärtlindring under influensan i fem dagar. Växten Echinacea purpurea och dess extrakt används som behandling av luftvägsinfektioner exempelvis hosta och vid inflammatoriska tillstånd, som exempelvis bronkit. Studier på olika extrakt av Echinacea purpurea har visat ökade halter av cytokiner som exempelvis TNF-α, IL-1 och IFN- β. Det har även visat en ökad aktivitet av fagocyterna och makrofager. En ökad mobilitet har också setts hos både leukocyter och aktivering av NK-celler. Syfte Syftet med arbetet var att undersöka om Echinacea purpurea har en symtomlindrande effekt på influensa A, speciellt på symtom som feber och hosta. Metod Publicerade vetenskapliga artiklar söktes i databasen Pubmed och OneSearch. Sökorden för artiklarna var “Influenza AND Echinacea purpurea” eller “Echinacea AND flu” I OneSearch användes sökorden “Influenza AND Echinacea purpurea”.ResultatStudierna visar att symtomen hosta och feber reducerades vid användning av Echinacea purpurea extrakt under behandling av influensa A. Behandlingen med Echinacea purpurea hade en bättre verkan vid användningen av en högre dos av extraktet (2000 mg) och om det användes några dagar innan influensasymtom uppträdde eller vid början av influensasymtom. Slutsats Utifrån de publicerade resultaten av de fyra studierna verkar Echinacea purpurea ha en mildrande effekt på hosta och feber under akuta luftvägsinfektioner. Det visade att dosen som hade mest effekt i dessa studier, var 2000 mg av växtextrakt från Echinacea purpurea. Dessa resultat erhölls vid behandling av i övrigt friskaindivider som endast hade förkylningssymtom eller influensasymtom, vilket gör att Echinacea purpurea extrakt kan ha andra effekter om det används på kroniskt sjuka individer. Flera äldre vaccinerar sig mot influensa, vilket gör att Echinacea purpurea extrakt har mer gynnsam effekt på yngre personer som har drabbats av influensa t.ex. barn eller unga vuxna. / Background Influenza is an acute respiratory infection which is caused by a negative stranded RNA-virus. Infection usually happens during the winter season. There are four different types of influenza, A-D. Type A is the most common one and affects both humans and animals, while type D has only been detected in pigs and cattle. Influenza starts with chills with a quickly rising fever (over 40℃) together with muscle pains. Afterwards the infection spreads to the airways, where the symptoms of dry cough arise. Influenzaspreads through airborne aerosols or through contact. The influenza virus enters the hostcell through the surface proteins HA and NA, while later entering the nucleus where the replication of the virus occurs. Before the flu season begins, people over the age of 65, pregnant women and patients with a chronic disease are vaccinated. This is done to prevent serious diseases that may occur during the flu and to lower the rate of secondary infections and also to prevent death. Medications such as oseltamivir and zanamivir can be given to treat the infection; however, they are mostly used to relieve symptoms. Echinacea purpurea is a traditionally used medicinal plant which has been used to treat respiratory tract infections like respiratory infections and inflammatory conditions such as bronchitis. Studies on different extracts of Echinacea purpurea have shown that they increase levels of TNF-ɑ, IL-1 and IFN-β as well as an increase of macrophages and other phagocytes after treatment with Echinacea extract. Similarly, the extract has shown an increased mobility of leukocytes and activation of Natural Killer-cells. Aim The aim of this study was to examine if treatment with medications containing extracts of Echinacea purpurea relieved symptoms such as, cough and fever, when used in the treatment of influenza A. Method The previously published articles were searched for by using the databases Pubmed and OneSearch. The words used in the search for published articles were “Influenza AND Echinacea purpurea” or ”Echinacea AND flu” in Pubmed. In the database, OneSearch the words “Influenza AND Echinacea purpurea'' were used. Results The selected studies showed that the symptoms such as cough and fever, were reduced in studied patients that used Echinacea purpurea extracts during the treatment of influenza A. Infection was verified through nasal swabs that then were screened. The results showed that the treatment was more effective when using a dose of 2000 mg of Echinacea purpurea extract. Moreover the treatment was effective when used a few days before symptoms occurred or at the beginning of a respiratory tract infection. Conclusion The results from the four studies suggested that the extract from Echinacea purpureareduces cough and fever during acute respiratory infections. The dosage of 2000 mg Echinacea purpurea extract had the most effect. However the extracts were tested on healthy people that only had common cold symptoms or influenza symptoms, thus different results could have been obtained if the effects were tested on unhealthy individuals.

La fabrication d’un médicament sous forme de comprimés à base d'une plante fraîche reste actuellement encore un véritable enjeu et ce, à cause de la variabilité qui peut exister dans les différents lots d'extraits de plantes ainsi qu’à la limite imposée par les techniques de fabrication de comprimés existantes actuellement. Différents problèmes rencontrés avec la formulation actuelle de ce type de comprimés ont été observés tels que : les propriétés physiques du comprimé (très faible dureté des comprimés et temps de désagrégation assez long), goût désagréable, grande variabilité (variabilité de l'extrait sec) et mauvaise stabilité de la substance médicamenteuse. Des observations antérieures ont indiqué que la stabilité de la substance médicamenteuse dans les comprimés d'Echinacea purpurea ainsi que le goût peuvent être améliorés en la mélangeant avec de la bêta-cyclodextrine (β-CD). Dans la thèse de doctorat, une formulation actuelle commercialisée de comprimés d’Echinacea purpurea a été réalisée par la technique de la granulation par voie humide, avec un mélangeur à cisaillement élevé. Dans la formulation, presque la totalité de l'excipient (lactose monohydraté) est mouillé par le concentré d'Echinacea purpurea. Afin de réduire la quantité d'excipients à granuler et à sécher et d’obtenir un produit avec moins cher des couts de matériaux premières, un procédé de granulation classique a été proposé où seulement une fraction de la quantité totale de charge (cellulose microcristalline (MCC)) est utilisée pour la granulation et le séchage; le reste de la charge (sorbitol) est ajouté après la granulation. Ce granulat peut servir de matériau de base des différents comprimés. Dans les différentes expérimentations réalisées, la teinture d’Echinacea purpurea a été utilisée comme modèle pour l'optimisation de la fabrication des comprimés à base de plantes. L'objectif de la thèse était par conséquent de développer une nouvelle formulation de comprimés d’Echinacea purpurea en utilisant un procédé de granulation classique. Avec une meilleure maîtrise de la granulation humide (WGP) et son influence sur les propriétés physiques des comprimés, ceux-ci doivent être optimisés du point du vue de la stabilité de l'ingrédient actif qui se présente sous forme solide (alkylamides) et des propriétés physiques des comprimés, en particulier le taux de dissolution et les propriétés physiques des comprimés. / One current formulation of Echinacea tablets which is examined in the present thesis is to produce tablets in a wet granulation process (WGP) with a high shear mixer. During the manufacturing, almost the whole amount of the excipient (lactose monohydrate) is wetted by Echinacea purpurea concentrate. In order to reduce the amount of excipients being granulated and dried by a basis granulate method was proposed where only a fraction of the total amount of filler (Microcrystalline cellulose, MCC) is used for granulation and drying, the rest of the filler (sorbitol) is added after granulation. This granulate can serve as basis material for different tablets.Purpose: in the PhD thesis, tablets containing Echinacea purpurea tincture were used as a model for the optimization of herbal tablets. The aim of the dissertation was to develop a new Echinaforce formulation based on the “Basis Granulate” technology. With deeper understanding of the WGP and its influence on the physical tablet properties, the new Echinaforce tablets should be optimized in term of cost of goods, taste of tablets, stability of the active ingredient in solid forms (alkylamides) and the physical tablet properties of Echinaforce tablets, especially the dissolution rate and the compaction properties of the final tablet.

As a potential bioremediation system for contaminated soils, I evaluated the use of an arbuscular mycorrhizal (AM) fungus, Glomus intraradices on roots and shoots uptake of polycyclic aromatic hydrocarbons (PAHs), alkyl PAHs, and toxic metals in Echinacea purpurea, in using a controlled 20-week greenhouse study and a complimentary 2-year field study. E. purpurea seeds were either inoculated with the mycorrhizal fungus (AM) or not inoculated (non-AM) and grown in soil provided by the National Capital Commission (NCC) that have known contamination. In the greenhouse study, AM inoculation increased the uptake of alkyl PAHs in the roots of E. purpurea. The AM inoculation showed no effect on root uptake of PAHs and toxic metals over the 20-week study period. However, when I calculated the uptake rates (k1) for PAHs between both treatments, the AM treated roots ha 10-fold higher k1 values than non-AM treated roots. The soil concentrations of PAHs were found to increase over time with AM inoculation, suggesting, that AM fungi are causing a solvent depletion through root uptake of minerals and carbon, which concentrates the more hydrophobic PAHs in soils. Alkyl PAHs and metals showed no change over time amongst any of the treatments. Assessing the performance of AM fungi on the uptake of contaminants under field conditions, only PAHs showed increased bioaccumulation in the shoots of E. purpurea with AM inoculation. Alkyl PAHs and metals in plant material were unaffected by the AM inoculation, but increased significantly from year 1 to year 2. The uptake rates among treatments were similar, with non-AM roots having slightly greater uptake. Soil concentrations of PAHs and alkyl PAHs were unaffected over the course of the experiment. Our control soil, however, showed significant increases in concentration from year 1 to year 2 with alkyl PAHs. These results quantified the influence of AM hyphae-mediated uptake of organic and inorganic contaminant transfer from soil to plants and the bioaccumulation kinetics for contaminants by E. purpurea that will be useful for environmental models and phytoremediation strategies.

Rausvažiedė ežiuolė (Echinacea purpurea (L.) Moench) apibūdinama kaip priemonė skatinanti medžiagų apykaitą; stimuliuojanti metabolizmo pakitimus ir ląstelių natūralią gynybą esant lėtinėms ir ūmioms ligoms; antiseptikas; dezinfekuojantis žaizdas agentas; vaistas. Darbo tikslas buvo atlikti fenolinių junginių analizę rausvažiedės ežiuolės (Echinacea purpurea (L.) Moench) ekstraktuose, gautuose skirtingais metodais, naudojant spektrofotometrinį metodą ir efektyviąją skysčių chromatografiją. Darbo uždaviniai buvo optimizuoti ekstrakcijos sąlygas (maceracijos metodu) įvertinant skirtingas metanolio koncentracijas ir ekstrakcijos trukmės įtaką fenolinių junginių kiekybinei sudėčiai ekstraktuose; paruošti ekstraktus maceracijos, soksleto ir ekstrakcijos superkritiniais skysčiais metodais; optimizuoti efektyviosios skysčių chromatografijos sąlygas; atlikti kokybinę ir kiekybinę fenolinių junginių analizę Rausvažiedės ežiuolės ekstraktuose, įvertinant a) skirtingas ekstraktų paruošimo metodikas; b) skirtingas augalo dalis. Tyrimo metu optimizuotos ekstrakcijos sąlygos (maceracijos metodu); nustatyta, kad daugiausiai fenolinių junginių išekstrahuota naudojant 70 % metanolį per pirmas 4 ekstrakcijos valandas. Paruošti ekstraktai maceracijos, soksleto ir ekstrakcijos superkritiniais skysčiais metodais. Spektrofotometru nustatyta, kad daugiausiai fenolinių junginių išekstrahuojama maceracijos metodu — 3 kartus daugiau nei soksleto metodu ir 14 kartų daugiau nei ekstrakcija... [toliau žr. visą tekstą] / Echinacea purpurea (L.) Moench (family Asteraceae) herbal medicines and dietary supplements are traditionally used as immunostimulants in the treatment of inflammatory and viral diseases. Employed are roots and aerial parts. The main active compounds of Echinacea purpurea (L.) Moench are alkamides and polyacetylenes, caffeic acid derivatives, polysaccharides and glycoproteins. With regard to caffeic acid derivatives, several compounds have been identified from the hydrophilic fractions of Echinacea extracts, such as caftaric acid, chlorogenic acid, caffeic acid and cichoric acid. Cichoric acid is found to be the main phenolic compound in E. purpurea. From the caffeic acid derivatives, only cichoric acid has shown immunostimulatory properties, promoting phagocyte activity in vitro and in vivo. In addition, cichoric acid has antihyaluronidase activity, and has a protective effect on the free-radical-induced degradation of collagen. Cichoric acid has also shown antiviral activity and has recently been found to inhibit HIV-1 integrase and replication. Tasks of investigation: to prepare samples for phenolic acids analysis by supercritical fluid, soxhlet extractions and maceration methods and optimize SPE parameters. Evaluate amount of cichoric acid and compare it in different parts of plant by spectrophotometric method. Optimize HPLC analysis conditions and identify the main phenolic acids by HPLC. Compare amounts of phenolic acids between plant parts and extraction methods... [to full text]

Vaistiniai augalai šiuolaikinėje medicinoje turi labai didelę reikšmę. Šiuo metu Lietuvoje – tradicinėje ir liaudies medicinoje naudojama apie 460 augalų rūšių. Rausvažiedė ežiuolė yra vienas iš populiariausių vaistinių augalų. Iš jos pagaminti vaistai vartojami odos, onkologinių ir peršalimo ligų gydymui. Vaistinei žaliavai naudojamos įvairios rausvažiedės ežiuolės dalys: lapai, žiedai, stiebai, šaknys ir t.t. Nuimta vaistinė augalinė žaliava yra labai drėgna. Susidaro ypač palanki terpė mikroorganizmams vystytis. Todėl vaistinės augalinės žaliavos paruošimui dažniausiai naudojamas džiovinimas. Džiovinimo būdai didžia dalimi nulemia vaistinės augalinės žaliavos kokybę. Tyrimų tikslas – ištirti ir palyginti džiūvimo sąlygų įtaką rausvažiedės ežiuolės žolės ir skirtingų jos dalių drėgmės mainams su aplinka. Nustatyta, kad standžios augalo morfologinės dalys lemia stabilesnę ir poringesnę vaistinės augalinės žaliavos sampilo struktūrą, todėl džiovinant rausvažiedės ežiuolės žiedus ir ypač smulkintus stiebus bei žolę susiformuoja palankesnės džiūvimui sąlygos nei, pavyzdžiui, skintų lapų sampile. / Medicinal plants are of great importance in modern medicine. Currently, Lithuania - traditional and folk medicine used for about 460 species of plants. Echinacea purpurea is one of the most popular medicinal plants. From these plants made drugs are used to treat skin diseases, oncological diseases and cold diseases. Various parts of Echinacea purpurea an used for medicinal materials: leaves, flowers, stems, roots, etc. Been harvested herbal ingredients are very moist. There is a particularly favorable environment for the development of microorganisms. Therefore, the preparation of medicinal plant raw materials commonly used in drying. Drying methods are largely determined by the quality of raw materials. Purposes of the study - to investigate and compare the drying conditions on Echinacea purpurea herbs and different parts of moisture exchange with the environment. It was found that the rigid parts of the plant determines the morphological stability and more porous medicinal plant raw materials layer structure, therefore drying purple cone flowers, and especially the chopped stems and grass formed favorable drying conditions than cutted leaves layer.

Echinacea purpurea belongs to adaptogenic plants containig pharmacologically active compounds. The most important parts of the plants for medical use are roots, but also aboveground parts are processed. The plants were grown in triennial precise small-parcel experiment using acetylsalicylic acid as an in vivo elicitor to increase the content of bioactive substances. The contents of cichoric acid, caffeic acid, chlorogenic acid and caftaric acid were assayed in roots, tops and flowers. Repeated application elicitors on leaves were conducted with three different concentrations in fortnightly intervals. Ethanol extracts from the plant samples were analysed by high performance liquid chromatography. The effect of the elicitor on the content of bioactive substances was statistically significant. The optimal concentration of the elicitor considering the content of bioactive substances in roots is the middle one. The effect of elicitor is year-dependent.

THE ASSESSMENT OF INFLUENCE OF ZINC IONS AND ECHINACEA PURPUREA (L.) MOENCH FOR MICE INTOXICATED BY CADMIUM IONS Abstract Background. Cadmium (Cd), a well-known environmental hazard, exerts a number of toxic effects in organism. It disturbs the activity of biochemical systems of cells. Accumulation of cadmium depends on the amount of essential trace elements, including zinc. Echinacea purpurea (L.) Moench (EP) can modify its influence. The aim of the study was to assess the influence of ions of cadmium, zinc, and EP on organism in experimental model of mice. The objectives of the scientific work were as follows: 1. To evaluate the accumulation of cadmium in the internal organs of experimental mice after acute and chronic intraperitoneal and per oral intoxication. 2. To assess morphological changes in liver tissue, mitotic and apoptotic activity of liver cells after the intoxication by cadmium ions of different duration and dose. 3. To assess the effect of zinc ions to the accumulation of cadmium in the internal organs and to the mitotic and apoptotic activity of liver cells in the organism of mice intoxicated by cadmium. 4. To evaluate the effect of EP to accumulation of cadmium in internal organs, mitotic and apoptotic activity of liver cells after the chronic intraperitoneal and per oral intoxication by cadmium ions. The scientific novelty of the study. This work makes our knowledge about mechanisms and outcomes of acute and chronic exposure to cadmium deeper. The... [to full text]