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.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Abnormal movements are clinical symptoms present in neurodegenerative diseases, such Parkinsonism , as well as Tardive dyskinesia. It is thought that unbalance in monoamine levels with their consequent metabolism could be involved in the etiology of these abnormal movements. However, until this moment there is not efficacious treatment with low side effects for these conditions. In this context, Hypericum perforatum (H. perforatum), popularly known as St. John s Wort, is a plant largely used as antidepressant and presenting high amount of polyphenol constituents. Its antidepressant mechanism seems to involve the increase of monoamines and monoaminoxidase inhibition. Here, we evaluated the effect of H. perforatum on different models of abnormal movements in rats, either using fluphenazine or reserpine. It were quantified the number of vacuous chewing movements (VCMs) and locomotor activity (number or rearings and crossings) in both models. In experiment 1, rats received a single administration of fluphenazine enantate (25 mg/Kg, i.m.) and/or H. perforatum (300 mg/Kg, in place of drinking water) during 7 days. Fluphenazine treatment increased VCMs and decreased the locomotor activity after 7 days of treatment. However, H. perforatum did not alter either the number of VCMs or the locomotor activity (represented by number of crossing and rearing in the open field test) in rats. In experiment 2, rats received reserpine administration once a day during 3 days (0.5 mg/Kg, s.c.) and/or H. perforatum (300 mg/Kg, in place of drinking water) during 16 days. Reserpine treatment increased VCMs and decreased the locomotor activity. H. perforatum did not alter the number of VCMs or the number of rearing. However, H. perforatum co-treatment could avoid the effect of reserpine on number of crossings. In conclusion, H. perforatum did not seem to be efficacious to protect against orofacial movements induced by fluphenazine or reserpine in rats. / As desordens motoras são sintomas de doenças neurodegenerativas bem como podem estar associadas ao tratamento com antipsicóticos. Cita-se como exemplo destas desordens, sintomas da Doença de Parkinson (DP) e a Discinesia Tardia (DT), respectivamente. O desequilíbrio cerebral nos níveis de monoaminas e, consequentemente, seu metabolismo tem sido relacionados ao desenvolvimento dos movimentos anormais que aparecem nas desordens motoras, tendo em vista que os circuitos dopaminérgicos estão envolvidos na gênese da DP e da DT. No entanto, ainda não existem tratamentos eficazes e com poucos efeitos colaterais para tais condições. A Hypericum perforatum (H. perforatum), popularmente conhecido como Erva de São João, consiste numa planta amplamente utilizada como antidepressiva e que possui uma grande quantidade de compostos fenólicos, cujo mecanismo para esta atividade está relacionado à inibição da monoamina oxidade (MAO) e da recaptação de monoaminas cerebrais. Assim, o objetivo do presente estudo foi investigar o efeito do H. perforatum em modelos de desordem motora induzida por flufenazina ou reserpina em ratos. Foram quantificados os movimentos de mascar no vazio (MMV) e atividade locomotora (cruzamentos e levantadas observados no teste de campo aberto) em ambos os modelos. No experimento 1, os ratos receberam uma única administração de enantato de flufenazina (25 mg/Kg, i.m.) e ou H. perforatum (300 mg/Kg, no lugar da água de beber) durante 7 dias. O tratamento com flufenazina aumentou o número de MMVs e diminuiu a atividade locomotora em ratos após 7 dias de tratamento. No entanto, o tratamento com H. perforatum não protegeu das alterações comportamentais causadas pelo tratamento com flufenazina. No experimento 2, os ratos receberam água ou H. perforatum (300 mg/Kg, no lugar da água de beber) durante 16 dias. A partir do dia 9 de tratamento os animais receberam uma administração diária de reserpina (0,5 mg/Kg, s.c.) ou veículo durante 3 dias com intervalo de 48 horas. A reserpina aumentou o número MMVs e diminuiu a atividade locomotora em campo aberto. O prétratamento com H. perforatum não alterou o efeito da reserpina sobre o número de MMVs e levantadas. Porém, o pré-tratamento com H. perforatum preveniu o efeito da reserpina sobre número de cruzamentos. Desta forma, podemos concluir que o H. perforatum não apresentou efeito benéfico sobre os movimentos orofaciais induzidos por flufenazina ou reserpina em ratos.