Hypericum polyanthemum é uma planta nativa do Sul do Brasil que contém compostos como flavonóides, taninos, derivados de floroglucinol (uliginosina B) e benzopiranos: HP1 (6-isobutiril-5,7-dimetóxi-2,2-dimetilbenzopirano), HP2 (7-hidróxi-6-isobutiril-5- metóxi-2,2-dimetilbenzopirano), e HP3 (5-hidróxi-6-isobutiril-7-metóxi-2,2- dimetillbenzopirano). Esses metabólitos são responsáveis por uma série de atividades biológicas como inibidores da monoaminoxidase (IMAO), antiproliferativa e antitumoral. No presente trabalho modificações na biomassa vegetal e no teor de metabólitos bioativos de plantas de H. polyanthemum cultivadas sob condições controladas e após 18 semanas de aclimatação foram investigadas após elicitação com o fungo Nomuraea rileyi através da adição do microrganismo liofilizado e pulverizado (LP), ou liofilizado, autoclavado e pulverizado (LAP) por curtos e longos períodos de tempo. Plantas cultivadas sob condições controladas tratadas com LAP demonstraram aumento na concentração dos benzopiranos HP1, HP2 e HP3 enquanto LP provocou efeito negativo ou ainda, não alterou a síntese destes compostos. Porém, baixos níveis do floroglucinol uliginosina B foram detectados em todos os tratamentos. Tratamentos com LAP por longos períodos de tempo em plantas cultivadas a campo provocaram aumento de biomassa (2x) e a análise química demonstrou elevação do teor de compostos fenólicos totais nas partes vegetativas das plantas submetidas a todos os tratamentos. Ainda, a análise apontou um diferente grau de acúmulo dos metabólitos, com maiores teores de HP1, HP2, HP3 e uliginosina B sendo encontrados nas partes reprodutivas das plantas tratadas com LAP. A elevação dos metabólitos bioativos em resposta ao elicitor fúngico sugere que estes compostos são induzíveis na resposta de defesa de H. polyanthemum. Os resultados obtidos nesse trabalho são relevantes em virtude da planta não sofrer danos. Assim, o sistema descrito representa uma nova abordagem em pesquisas com plantas visando otimizar condições para a produção de biomassa e metabólitos secundários de interesse medicinal. / Hypericum polyanthemum is a native plant of South Brazil that contains compounds such as flavonoids, tannins, phloroglucinol derivatives (uliginosin B) and benzopyrans: HP1 (6-isobutyryl-5,7-dimethoxy-2,2-dimethylbenzopyran), HP2 (7-hydroxy-6-isobutyryl-5- methoxy-2,2-dimethylbenzopyran), and HP3 (5-hydroxy-6-isobutyryl-7-methoxy-2,2- dimethylbenzopyran). These metabolites are responsible for biological activities such as monoamine oxidase inhibitor, antiproliferative and antitumoral. In this work changes in biomass and bioactive metabolites after elicitation with the fungi Nomuraea riley added as freeze dried culture (DC) or as freeze dried autoclaved cell powder (DACP) for short and long periods of time have been investigated in Hypericum polyanthemum plants grown under controlled conditions and after 18 weeks of field acclimatization. Plants treated with DACP showed increased concentrations of the benzopyrans while the DC affected negatively or did not alter the synthesis of these compounds. Nevertheless, low levels of uliginosin B were detected in all treatments. Long time treatment of field grown acclimatized plants with DACP triggered plant growth doubling the biomass and chemical analyses demonstrated increased total phenolic compounds yields in the vegetative parts of plants submitted to the treatments. Furthermore, the analysis showed different pattern of metabolites accumulation, with higher yields of benzopyrans and uliginosin B accumulated in the reproductive parts of the plants treated with DACP during all experiment. The elevation of bioactive metabolites levels in response to the elicitor suggests that these compounds are inducible in plant defense response of H. polyanthemum. The results obtained are relevant since the plant did not suffer injury. Then, the system described represents a new approach in plant research aimed to optimize the conditions for biomass and accumulation of secondary metabolites.

The main topic of this work is food supplements, their legislation and other theory connected with them, the main focus is on plant nutraceuticals, of which are chosen food supplements marketed to support immunity system - echinacea, sea buckthorn and oyster mushroom, food supplements to support psyche - St. John's wort, ginkgo and valerian and dietary supplements to support body fitness - tribulus, garcinia and psyllium. Subsequently, the topic focuses on the marketing of nutraceuticals, its modern ways and the possibilities of increasing the students' awareness in this field through the principles of media education. In the practical part, 112 students aged 15-19 are presented with a lecture presenting the topic and inviting students to raise their interest in the subject. The feedback of the lecture is provided by a questionnaire whose questions correspond to the hypotheses set by the author. KEYWORDS nutraceuticals, media education, marketing, Echinacea, Garcinia, Pleurotus, Hippophae, Gingko, Plantago, Hypericum, Valeriana, Tribulus

Abstract Hypericum perforatum L. (St. John’s wort) is a medicinal plant widely utilized for the treatment of depression. The antidepressant activity is mainly attributed to the phenolic compounds hypericins and hyperforins, which also have a wide range of other pharmacologically interesting properties. The biosynthetic routes leading to hypericins and hyperforins are poorly understood, although a polyketide pathway including type III polyketide synthases (PKSs) has been suggested to be involved. Furthermore, a gene called hyp-1 is assumed to attend to the final stages of the hypericin biosynthesis. In the present work, the biosynthesis of hypericins and hyperforins in H. perforatum was further studied by focusing on the elucidation of the precursors and genes involved. The incorporation of isotopically labelled branched-chain amino acids into hyperforins was investigated as well as the possibilities to enhance the production of hyperforins in H. perforatum in vitro cultures by feeding them with amino acid precursors. Furthermore, two novel cDNAs encoding for type III PKSs were isolated from H. perforatum. The functions of these new genes, designated HpPKS1 and HpPKS2, as well as the role of hyp-1 were elucidated by comparing their expression with the levels of hypericins and hyperforins in H. perforatum tissues. The enzymatic activity of the recombinant HpPKS2 protein was also analyzed. To study Hyp-1 at a protein level, a protein extraction method was optimized for tissues of Hypericum species. The results show the incorporation of valine and isoleucine into the acyl side chain of hyperforin and adhyperforin, respectively. Through the biotransformation of the amino acid precursors, it is possible to enhance the levels of adhyperforin, but not hyperforin, in H. perforatum shoot cultures, which demonstrates the tight regulation of the hyperforin biosynthesis. A correlation between HpPKS1 expression and hyperforins was detected in H. perforatum tissues. The localization of HpPKS2 mRNA in dark glands in which hypericins accumulate as well as the octaketide synthase activity of the recombinant HpPKS2 suggest that HpPKS2 is associated with possible co-operating tailoring enzymes in the biosynthesis of hypericins. The presence of both hyp-1 mRNA and Hyp-1 protein in distinct places compared with hypericins in H. perforatum tissues does not support the idea that Hyp-1 would be involved in the biosynthesis of hypericins in dark glands, although mobility of the Hyp-1 protein was shown to be possible. The present thesis extends knowledge about the biosynthesis of hypericins and hyperforins in H. perforatum by providing new candidate genes for their biosynthesis and by identifying precursors for hyperforins. Moreover, new information was obtained about the role of hyp-1 in H. perforatum.

The main purpose of the theses was to find if the abiotic elicitor 2,4,6-trimethyl-N-(pyrazine- 2-yl)benzenesulfonamide has any influence on the secondary metabolites production in callus and suspension cultures of Hypericum perforatum L. The cultivation was taking place in the Murashige a Skoog (MS) nutrient medium enriched by the growth regulator - alpha-naphthyl acetic acid at the concentration of 1 mg.L-1 . The elicitor was added to the cultures at the three levels of concentration: c1= 100.0 mg/100 ml; c2= 10.0 mg/100 ml; c3= 1.00 mg/100 ml. The individual samples were taken after 6, 24, 48, 72 and 168 hours of the elicitor application. The control samples without the elicitor were taken after 6, 48 and 168 hours. The collected samples were dried and subsequently transformed into methanol extracts in order to determine secondary metabolites content (rutin, hyperoside and quercetin) by HPLC method. Release of these metabolites into nutrient media was also subject of this observation. The elicitation has influenced production of the secondary metabolites, particularly in callus cultures, wherein several statistically significant values, characterizing increase in their production, were measured. The highest content of rutin (0.169 mg.g-1 DW) was determined in callus culture after 168 hours when...

An immediate demand exists for new tuberculosis (TB) antibiotics due to the ever-increasing spread of drug-resistant strains. The drug-development process goes through four phases, the first (Phase 0) of which is to demonstrate and investigate drug effectiveness and toxicity. This research investigated the effectiveness of the Hypericum perforatum herb (commonly St. John's wort (SJW)) in its growth inhibition of mycobacteria and its viability effect on human lung cells. Organic-solvent SJW extracts were effective at inhibiting every nonpathogenic genetically sequenced Mycobacterium isolate currently available (six isolates) in preliminary studies. Quantitative studies of five Mycobacterium isolates showed an order of concentration sensitivity to the SJW methanol (MeOH) extract as (high to low) M. JLS, M. KMS, M. phlei (not sequenced), M. MCS, B. subtilis, M. smegmatis, and E. coli, with minimal bactericidal concentrations (MBCs) ranging from 0.33-2.66 mg extract/ml. The SJW compounds hyperforin (Hfn), hypericin (Hpn), and pseudohypericin (Phn) were quantified using a novel HPLC method that utilized common HPLC equipment. A crude MeOH extract solution of 133 mg extract/ml contained 2.26 mg Hfn/ml, 0.77 mg Hpn/ml, and 2.67 mg Phn/ml. Purified Hfn had a MBC of between 6-13 μg/ml for M. JLS in the absence of Tween 80. Tween 80 repressed Hfn (46 μg/ml) inhibition of M. JLS at ≥ 0.025% (v/v). Purified Hpn and Phn showed no inhibition of M. JLS at all assayed concentrations, which were ≤ 27 μg/ml and ≤ 25 μg/ml, respectively. Inhibitory results from the five quantitatively assayed Mycobacterium samples could be extrapolated to M. tuberculosis, as these isolates have as high as 72% genetic homology to the pathogen. The crude MeOH extract and Hfn were lethal to the human carcinomic alveolar epithelial lung cell line A549 at 1.3 mg extract/ml (crude extract) and ≥ 11 μg/ml (Hfn), with a Hfn LD50 of 3-6 μg/ml (5.6-11.2 μmol/L). Because Hfn is antiproliferative to a list of other carcinomic cell lines in the same concentration range, the A549 cell line may be added to that list. The addition of M. JLS cells (5x106 cells/cm2) to penicillin-streptomycin-containing A549 culture (which killed the bacteria) did not affect A549 viability.

This was a chemical laboratory study. The main focus was to evaluate the chemical stability of Hypericum perforatum (St John's wort), Ginkgo biloba and Piper methysticum (Kava Kava) under unfavourable environmental conditions. Different dosage forms representing the same amount of active ingredients for each were used. Some of the dosage forms were self manufactured according to Good Manufacturing Practice. Samples of the dried powder of each plant was also exposed to a series of gamma¬radiation. Acetone was used as an extractant for all three plants, after evaluating and discarding the extraction method stipulated in the British Herbal Pharmacopoeia. Identification of the different plants were carried out by means of Thin Layer Chromatography. The in-house developed mobile phases EMW, BEA and CEF, showed better separation and visibility compared to the mobile phases used in the British Herbal Pharmacopoeia. The plates were sprayed with either vanillin or p-anisaldehide for optimal visualization of the separated compounds. After the specified period of 6-months, comparative TLC was performed on all samples. This was achieved for each plant by applying all samples stored at a specific condition i.e.25°C, on the same plate. The samples were stored at low temperature after exposure to the specific time interval. Quantitative analysis was performed by spectrophotometry, and high pressure liquid chromatography. The data obtained from these analytical methods, were used to evaluate the relative chemical stability of each dosage form. The relationship between the quantitative data and the qualitative changes in the TLC fingerprints, were compared, hoping to achieve a common pattern relating to the stability. The order of the reaction as well as the reaction rate constant (k) for each dosage form was calculated, except for kava kava. The shelf-life (too) was calculated using the analyzed data obtained by spectrophotometry or HPLC. The relevance of conventional pharmaceutical calculations in the prediction of shelf-life, by means of accelerated stability tests, was investigated for the possible application to herbal products. The effects of gamma radiation on the degradation of the chemical compounds present in each plant, was evaluated. After an evaluation of all the relevant data, it seemed that the tablet-dosage forms were equally effective regarding stability, compared to the capsules. Liquid extracts appeared to be less stable than the extract capsules. The extract capsules seemed to degrade more rapidly than the herbal tablets or herbal capsules. Exposure to low dose radiation (4.4 kGy) did not seem to have an influence on the stability. It was evident that some herbs were more sensitive to sunlight or heat than others. In general, all three of the chosen plants seemed to be relatively stable if stored in the specified conditions. It seemed valid for the shelf-life to be expressed as two years. / Dissertation (MSc (Pharmacology))--University of Pretoria, 2001. / Pharmacology / unrestricted

Antibiotic resistance is a rapidly increasing and serious global problem, with infectious diseases currently being the second leading cause of death worldwide. The World Health Organization (WHO) has estimated that 700,000 people worldwide die each year as a result of antibiotic resistance, and this number is projected to rise to 10 million per year by 2050 if no action is taken. This makes it vital to find new antibacterial agents to treat infectious diseases. Staphylococcus aureus (S. aureus) is a commonly occurring gram-positive bacterium from the genus Staphylococcus. The bacterium can cause several diverse types of infectious diseases, ranging from mild to life-threatening. Hypericum perforatum L. (H. perforatum) is a plant that has been used as a medicine in various preparations for thousands of years to treat different conditions, such as e.g., wound healing. Recent investigations have shown an antibacterial effect of the plant against the bacterium S. aureus, this suggested that H. perforatum may have a possible antimicrobial effect and could potentially be used in the treatment of certain infections. The purpose of this literature review was to investigate the in vitro antibacterial effect of H. perforatum against S. aureus. For this work, database searches were conducted in PubMed and SciFinder, and four studies were finally selected that examined the in vitro effect of H. perforatum against S. aureus. Common for all the studies was that they used the broth dilution method in their analyses to obtain MIC values. The results of the literature reviews showed that H. perforatum inhibited non-resistant isolates of S. aureus with MIC values between 0.01 – 950 µg/ml. For the resistant isolates evaluated, MIC values were between 0.01 – 0.02 µg/ml. The MIC-value against the multidrug resistant strain MDR-BAA 44 was 0.01 µg/ml. The other studies had higher MIC values and less activity compared to the results reported in the first study. The first study also showed that H. perforatum had better activity against all isolates than the positive controls tetracycline and penicillin. Further studies are needed to ensure and validate the results shown in study one. Also, research should focus on further studying the effect in vivo in larger studies than previously conducted. In addition, evaluation whether a synergistic effect against resistant S. aureus can be achieved through the combination of H. perforatum together with another plant species or with an antibiotic that could provide an alternative treatment of serious infections. / Infektionssjukdomar är idag den näst vanligaste dödsorsaken i världen och Världshälsoorganisationen (WHO) har beräknat att 700 000 personer i världen dör varje år till följd av antibiotikaresistens. Denna siffra kommer att stiga snabbt till 10 miljoner per år till år 2050 om inte åtgärder tas för att förhindra fortsatt spridning. Antibiotikaresistens är ett snabbt ökande och allvarligt globalt problem. Detta gör det livsnödvändigt att hitta nya antibakteriella medel för att kunna behandla infektionssjukdomar. Hypericum perforatum L. (H. perforatum) är en växt som i tusentals år använts som läkemedel iolika beredningar och för olika åkommor, som till exempel sårläkning. Antibakteriell effekt av växten har kunnat identifierats mot bakterien Staphylococcus aureus (S. aureus). S. aureus är en vanligt förekommande gram-positiv bakterie. Bakterien orsakar flera olika typer av infektionssjukdomar som kan vara allt från milda till livshotande. H. perforatum innehåller substanser med antibakteriell effekt som kan användas vid behandling av vissa infektionssjukdomar.  Syftet med denna studie var att undersöka den antibakteriella effekten in vitro av H. perforatum mot S. aureus. För detta litteraturarbete gjordes databassökningar i PubMed och SciFinder av tidigare publicerade studier. Fyra studier valdes slutligen ut som uppfyllde inklusionskriterierna. Samtliga studerade in vitro effekten av H. perforatum mot S. aureus. Samtliga studier använde sig av buljongspädningsmetoden i sina analyser där MIC-värden erhölls.  Resultaten från studierna visade att H. perforatum inhiberade icke-resistenta isolat av S. aureus med MIC-värden mellan 0,01 – 950 µg/ml. För de resistenta isolat som testades blev MIC-värden 0,01 – 0,02 µg/ml. MIC-värdet mot den multi-resistenta bakterien MDR-BAA 44 var 0.01 µg/ml. De andra studierna rapporterade högre MIC-värden jämfört med studie 1. Denna studie visade även att H. perforatum hade bättre aktivitet mot samtliga isolat än de positiva kontrollerna, tetracyklin samt penicillin. Vidare behöver fler studier genomföras för att säkerställa de resultat som studie 1 visade. Nya studier behöver också utvärdera om synergistisk effekt mot resistenta S. aureus kan uppnås vid kombination av H. perforatum och en annan växtart eller antibiotikum som kan leda till nya behandlingsalternativ av svåra infektioner.

No / Three new xanthones, hypericorin C (1), hypericorin D (2) and 3,4-dihydroxy-5-methoxyxanthone (3), along with eight known compounds; 2,3-dimethoxyxanthone (4), 3,4-dihydroxy-2-methoxyxanthone (5), 3,5-dihydroxy-1-methoxyxanthone (6), 3-acetylbetulinic acid (7), 10H-1,3-dioxolo[4,5-b]xanthen-10-one (8), 3-hydroxy-2-methoxyxanthone (9), 3,4,5-trihydroxyxanthone (10) and betulinic acid (11) were isolated from the roots of Hypericum oblongifolium. The structures of the new compounds 1, 2 and 3 were deduced by spectroscopic techniques [ESI MS, (1)H NMR, (13)C NMR, and 2D NMR (HMQC, HMBC, COSY and NOESY)]. The entire series of compounds were evaluated for anti-ulcer activity.

Made available in DSpace on 2014-12-17T14:13:22Z (GMT). No. of bitstreams: 1 SebastiaoDSF.pdf: 475002 bytes, checksum: d28adb2821adc67501d7e19f42e9b6a3 (MD5) Previous issue date: 2007-08-16 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico / Several clinic evaluations have been possible with radiobiocomplexes labeled with technetium-99m (99mTc). Some natural and synthetic drugs are capable of to interfere on the labeling of blood constituents with 99mTc, as well as on the biodistribution of radiobiocomplexes. Authors have also reported about the toxicity of several natural products. The aim of this study was to compare the effects of the Mentha crispa (hortel?) and of the Hypericum perforatum (hip?rico) in different experimental models. On the labeling of red blood cells (RBC) and plasma and cellular proteins with 99mTc, both extracts were capable of to decrease the radioactivity percentage on the cellular compartment and on the fixation on plasma and cellular proteins. On the morphometry of the RBC, only the hortel? was capable to alter the shape and the perimeter/area ratio of the RBC. On the biodistribution of the radiobiocomplex sodium pertechnetate (Na99mTcO4), the hortel? increased the Na99mTcO4 distribution in the kidney, spleen, liver and thyroid, meanwhile the hip?rico decreased the Na99mTcO4 distribution in the bone, stomach, lungs and thyroid, and increased the Na99mTcO4 distribution in the pancreas. On the bacterial cultures survival, the hip?rico was capable of to protect the bacteria against the stannous chloride (SnCl2) effect. The hip?rico did not alter the topology of plasmidial DNA and did not protect the plasmidial DNA against the SnCl2 action. Probably, the effects presented by both extracts could be due to chemical compounds of the extracts that could alter the morphology of the RBC and the plasma membrane ions transport, and/or by phytocomplexes that could be formed with different effects dependent on the biological system considered / Avalia??es cl?nicas t?m sido poss?veis com radiobiocomplexos marcados com tecn?cio-99mTc (99mTc). Drogas naturais ou sint?ticas s?o capazes de interferir na marca??o de estruturas sangu?neas com 99mTc, assim como na biodistribui??o de radiobiocomplexos. Tamb?m tem sido descrita a toxicidade de v?rios produtos naturais. O objetivo deste estudo foi comparar o efeito dos extratos de Mentha crispa (hortel?) e de Hypericum perfloratum (hip?rico) em diferentes modelos experimentais. Na marca??o de estruturas sang??neas com 99mTc verificou-se que ambos os extratos foram capazes de diminuir a radioatividade no compartimento celular, nas prote?nas plasm?ticas e celulares. Na morfometria das hem?cias, apenas a hortel? foi capaz de alterar a forma e a rela??o per?metro/?reas das hem?cias. Na biodistribui??o do radiobiocomplexo pertecnetato de s?dio (Na99mTcO4) a hortel? aumentou a capta??o do Na99mTcO4 no rim, no ba?o, no f?gado e na tire?ide, enquanto que o hip?rico diminuiu a capta??o do Na99mTcO4 no osso, no est?mago, no pulm?o e na tire?ide, e aumentou no p?ncreas. Na sobreviv?ncia de culturas bacterianas o hip?rico foi capaz de proteger a bact?ria do efeito danoso do cloreto estanoso (SnCl2). O hip?rico n?o alterou a topologia nem protegeu o DNA plasmidial da a??o do SnCl2. Provavelmente os efeitos apresentados por ambos os extratos poderiam ser explicados por subst?ncias presentes nos extratos que poderiam alterar a morfologia das hem?cias, o transporte de ?ons pela membrana e/ou formar fitocomplexos. O estudo teve car?ter multidisciplinar com a participa??o das seguintes ?reas do conhecimento: Radiobiologia, Bot?nica, Endocrinologia, Fitoterapia e Hematologia

Herbal medicines are growing in popularity in the Western world and are becoming more stringently regulated under new EU legislation. Within the arena of herbal medicines, St. John’s Wort (SJW), Hypericum perforatum, is a top ten best seller with clinical evidence to support its use as an anti-depressant. A fundamental requirement of the new legislation is to prove the identity of the plant material in question. This is currently achieved via morphological and chemical methods, neither of which are ideal. A wide range of DNA based methods have been applied to this arena, standardisation is required to realise the potential of DNA based techniques. The DNA barcoding initiative aims to produce sequence data for all plant species, capable of species identification. The proposal is to use these data to design fast and effective DNA based methods of identification. For assay design, the putative barcode region nrITS was selected as a platform. Three assays were designed; • A PCR assay designed to hyper variable sequences within a barcode region. This assay is capable of distinguishing SJW from other closely related species. • A quantitative qPCR assay designed to measure total DNA and specific SJW DNA within a mixed sample. • A multiplex PCR incorporating fluorescently labelled primers, allowing amplicon detection by capillary electrophoresis. This assay identifies four separate Hypericum species, including SJW, with a mixed sample in one reaction. The suitability of the nrITS and three other barcode regions is assessed based on sequence data generated for 32 vouchered samples of different Hypericum species, and a Lithuanian sample set of 22 and 16 H. perforatum and H. maculatum samples respectively. The matK is currently unusable, the rbcL highly conserved, trnH-psbA problematically variable and the nrITS proved to be ideal for assay design.