Optimization of Phenolics and Flavonoids Extraction Conditions and Antioxidant Activity of Roasted Yerba-Mate Leaves (Ilex paraguariensis A. St.-Hil., Aquifoliaceae) using Response Surface Methodology.

This study focused on maximizing the extraction of total phenolics and flavonoids as well as the antioxidant activity measured by the 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay from roasted yerba mate (Ilex paraguariensis) as a function of time (5, 7.5 and 10 min) and temperature of extraction (60, 75 and 90°C). The data were subjected to Response Surface Methodology and the results showed that polynomial equations were significant, did not present lack of fit, and presented adjusted determination coefficients above 98%, proving their suitability for prediction purposes. Using the desirability function, the optimum operating conditions to obtain a higher extraction of antioxidants was found to be 10 min of extraction at 90°C, and the tea prepared under these experimental conditions presented 427.74 mg of gallic acid equivalents per liter and 80.02% of inhibition of the DPPH radical. The flavonoid content was highly correlated (r = 0.9046, p < 0.001) to the antioxidant capacity.

Antidepressant profile of Ptychopetalum olacoides Bentham (Marapuama) in mice.

Depression has become of universal major importance, and it is therefore vital to expand the armamentarium for treating the condition. Lack of motivation and lassitude are major symptoms treated with the use of Marapuama (Ptychopetalum olacoides, PO) remedies by communities in the Brazilian Amazon. Considering the prominence of such symptoms in depression, the present study was designed to verify the effects of a standardized PO ethanol extract (POEE) on the forced swimming (FST) and tail suspension tests (TST). POEE i.p. (15-100 mg/kg) and oral (300 mg/kg) resulted in a significant and dose-related anti-immobility effect. We further examined the involvement of dopamine, noradrenaline and serotonin in these antidepressant-like effects. POEE effects were prevented when catecholamine synthesis was inhibited by -alpha-methyl-rho-tyrosine (AMPT) (100 mg/kg, i.p.), while inhibition of serotonin synthesis with rho-chlorophenylalanine methyl ester hydrochloride (PCPA) (100 mg/kg, i.p.) was devoid of effect. The blockade of beta-adrenergic (propranolol 2 mg/kg, i.p.) and D(1) dopamine (SCH 23390 0.5 mg/kg, i.p.) receptors prevented POEE anti-immobility effects; by contrast, blockade of D(2) dopamine (sulpiride 2 and 50 mg/kg, i.p.) receptors was ineffective. Consistent with traditional use, the results indicate that POEE possesses antidepressant-like effects, possibly mediated by beta-adrenergic and D(1) dopamine receptors.

Serotonin receptors contribute to the promnesic effects of P. olacoides (Marapuama).

Nootropic, antioxidant, and neuroprotective properties have been shown in a standardized ethanol extract of Ptychopetalum olacoides (POEE), a medicinal plant traditionally used by the Amazonian elderly population. It has been revealed that POEE mechanisms of action include anticholinesterase effects, and involve beta-adrenergic and dopamine D(1) receptors. The purpose of this study was to verify the role of serotonin receptors in the promnesic effects of this standardized extract. The step-down task in mice and selective serotonin antagonists were used. The study reveals that POEE promnesic effects on short-term (acquisition, consolidation and retrieval) and long-term (retrieval) declarative aversive memories are increased by 5HT(2A) (but not 5HT(1A)) serotonin antagonists (spiperone and pindolol, respectively). The observed synergism between POEE and spiperone can be interpreted as the combined effects of two subeffective doses of two 5HT antagonists, or the known synergism between an acetylcholinesterase inhibitor (POEE) and a 5HT antagonist. In conclusion it is suggested that 5HT(2A) serotonin receptors are relevant for the promnesic effects of this extract, adding to its multiple mechanisms of action.

Effects of Marapuama in the chronic mild stress model: further indication of antidepressant properties.

ETHNOPHARMACOLOGY RELEVANCE: Ptychopetalum olacoides Bentham (PO) (Olacaceae), known as Marapuama, is regarded as a "nerve tonic" in the Amazon. Traditional uses include states of lassitude with noticeable lack of desire/motivation, and to manage particularly stressful (physical and/or psychological) circumstances. Suggestive of antidepressant activity, we have established that a specific PO ethanol extract (POEE) significantly decreases immobility in the tail suspension and forced swimming tests. AIM OF THE STUDY: The aim of this study was to verify the effects of POEE in the unpredictable chronic mild stress (UCMS) depression model in mice, given the construct and face values of the UCMS as an experimental model of depression and the traditional use of this species. MATERIALS AND METHODS: Over 6 weeks BALB/c mice were subjected to the UCMS protocol. The effects of POEE (50, 100, 300mg/kg, p.o.) and imipramine (20mg/kg, i.p.) were evaluated in relation to coat state, splash-test grooming, and corticosterone levels. RESULTS: The coat state degradation, decreased grooming and increased serum corticosterone induced by UCMS were prevented by POEE and imipramine treatments. CONCLUSION: In addition to supporting traditional claims and previously reported antidepressant properties for POEE, this study shows that POEE prevents stress-induced HPA hyperactivity.

Promnesic effects of Ptychopetalum olacoides in aversive and non-aversive learning paradigms.

Homemade remedies with Ptychopetalum olacoides (PO) roots are used by Amazonian peoples for treating various age-related conditions. We previously reported that Ptychopetalum olacoides ethanol extract significantly improved step-down inhibitory avoidance long-term memory in adult and reversed memory deficits in aging mice. Adding to previous data, this study shows that a single i.p. administration of Ptychopetalum olacoides ethanol extract (POEE 50 and 100 mg/kg) improved step-down inhibitory avoidance short-term memory (STM) 3 h after training in adult (2.5 month) mice; comparable results were obtained with POEE given p.o. at 800 mg/kg. Moreover, memory improvement was also observed in aging (14 months) mice presenting memory deficit as compared to adult mice. Furthermore, POEE (100 mg/kg) improved non-aversive memory systems in adult mice in an object recognition paradigm. Consistently with its traditional use this study add to previously reported data and reinforces that POEE facilitates memory processes. Although the acetylcholinesterase inhibitory properties described for this extract may be of relevance for improving memory processes, the molecular mechanism(s) underlying the memory improvement here reported needs further scrutiny.

Antiulcerogenic effect and cytotoxic activity of semi-synthetic crotonin obtained from Croton cajucara Benth.

Trans-dehydrocrotonin, the major diterpene isolated from the bark of Croton cajucara, has good antiulcerogenic activity which, however, is accompanied by toxic effects. On the basis of these results, a semi-synthetic crotonin, named 4SRC, was prepared to determine whether this substance has similar antiulcerogenic activity with lower or no toxicity. The natural crotonin was also isolated from the bark of C. cajucara but was not used due to the small amount obtained. The cytotoxic effect of semi-synthetic crotonin, expressed as cell viability, was assessed in (a) lung fibroblast cell line (V79) derived from Chinese hamsters, a system commonly used for cytotoxicity studies, and (b) rat hepatocytes isolated from male Wistar rats. After treatment, cell viability was determined by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide reduction (MTT reduction), total acid content and neutral red uptake assays. To evaluate V79 cell viability, different concentrations of semi-synthetic crotonin were incubated with the cells. To evaluate the antiulcerogenic effects of semi-synthetic crotonin (50, 100 and 200 mg/kg), we used the models of gastric ulcer induced by ethanol/HCl, stress, indomethacin/bethanechol, and ethanol in male Swiss mice and male Wistar rats. The substance had an IC(50)=500 microM in the neutral red uptake and MTT reduction tests and an IC(50)=200 microM in the nucleic acid content test. With regard to hepatocyte viability after treatment with semi-synthetic crotonin at different concentrations, semi-synthetic crotonin had an IC(50)=10-500 microM in the nucleic acid content and MTT reduction tests and an IC(50)=120 microM in the neutral red uptake test. In another experiment, V79 cells were incubated with the metabolites produced by hepatocytes treated with different concentrations of semi-synthetic crotonin. After a 4-h incubation, semi-synthetic crotonin had an IC(50)=500 microM in the MTT reduction and neutral red uptake tests and an IC(50)=370 microM in nucleic acid content test. The substance had significant antiulcerogenic activity in all models studied, suggesting the presence of a possible antisecretory effect combined with a cytoprotective effect. For this reason, the effect of semi-synthetic crotonin was also evaluated on biochemical parameters of gastric juice and gastric wall mucus, both obtained from pylorus-ligated mice. No significant differences were observed in these parameters between semi-synthetic crotonin-treated and control animals. The results obtained with semi-synthetic crotonin are promising, with a significant preventive effect against gastric ulcer induced by different agents. Our data also show that semi-synthetic crotonin was less toxic than dehydrocrotonin and that the cytotoxic effects decreases with the time that isolated hepatocytes were in culture.

Neuroprotective effects of Ptychopetalum olacoides Bentham (Olacaceae) on oxygen and glucose deprivation induced damage in rat hippocampal slices.

Alcoholic infusions of Ptychopetalum olacoides Bentham (PO, Olacaceae) are used in traditional medicine by patients presenting age associated symptoms and those recovering from stroke. The aim of this study is to evaluate the neuroprotective properties of PO ethanol extract (POEE) using hippocampal slices from Wistar rats exposed to oxygen and glucose deprivation (OGD, followed by reoxygenation). Mitochondrial activity, an index of cell viability, was assessed by the MTT assay; in addition, the free radicals content was estimated by the use of dichlorofluorescein diacetate as probe. The OGD ischemic condition significantly impaired cellular viability, and increased free radicals generation. In non-OGD slices, incubation with POEE (0.6 microg/ml) increased (approximately 40%) mitochondrial activity, without affecting free radicals levels. In comparison to OGD controls, slices incubated with POEE (0.6 microg/ml) during and after OGD exposure had significantly increased cellular viability. In addition, at this same concentration, POEE prevented the increase of free radicals content induced by OGD. In view of the fact that respiratory chain inhibition and increased generation of free radicals are major consequences of the ischemic injury, this study suggests that Ptychopetalum olacoides contains useful neuroprotective compounds and, therefore, deserves further scrutiny.

Memory retrieval improvement by Ptychopetalum olacoides in young and aging mice.

Amazonian peoples use traditional remedies prepared with Ptychopetalum olacoides (PO) roots for treating various age-related conditions. This study shows that a single intraperitoneally (i.p.) administration of Ptychopetalum olacoides ethanol extract (POEE, 50 and 100mg/kg) improved memory retrieval in step-down inhibitory avoidance (P

Analytical strategy coupled with response surface methodology to maximize the extraction of antioxidants from ternary mixtures of green, yellow, and red teas (Camellia sinensis var. sinensis).

This work aimed at using a simplex-centroid design to model the effects of green, yellow, and red tea mixtures (Camellia sinensis var. sinensis) on metal chelation activity, phenolic composition, antioxidant activity, and instrumental taste profile. The regression models that described the extraction of flavan-3-ols, o-diphenols, total phenolic compounds (TPC), free radical scavenging activity toward 2,2-diphenyl-1-picrylhydrazyl radical (DPPH), cupric ion reducing antioxidant activity (CUPRAC), and ferric reducing antioxidant power (FRAP) were significant, and data were fit satisfactorily (R(2) > 80%). A mixture of green and red teas had a synergism in CUPRAC and TPC, whereas a mixture of yellow and red teas had a positive effect on CUPRAC and DPPH. An optimization was performed to maximize the antioxidant activity and flavan-3-ol content and to render a tea with mild bitterness, and results showed that a mixture of 14.81% green, 56.86% yellow, and 28.33% red teas would be the most suitable combination of factors.

MK801- and scopolamine-induced amnesias are reversed by an Amazonian herbal locally used as a "brain tonic".

RATIONALE: Traditional remedies prepared from Ptychopetalum olacoides (PO) are used throughout the Amazon to alleviate age-related conditions. These formulas are mainly used by elders, and alleged effects may be related to the anticholinesterase properties identified in a standardized ethanol extract of this species [P. olacoides standardized ethanol extract (POEE)]. OBJECTIVES: To further characterize the potential of this extract for developing drugs useful to treat cognitive deficits, the effects of POEE on scopolamine (scop)- and MK801-induced amnesias (acquisition, consolidation, and retrieval) in mice were investigated. RESULTS: Scop (3.0 mg/kg, ip) significantly impaired memory (all three phases) in the step-down inhibitory avoidance protocol. As expected, MK801 (0.1 mg/kg, ip) was amnesic regarding acquisition and consolidation, but not retrieval. POEE (100 mg/kg, ip) reversed the scop-induced impairment in all three phases of long-term and short memories, whereas only the memory consolidation deficit was reversed with MK801-induced amnesia. CONCLUSIONS: This study complements previously reported promnesic properties of this plant extract and suggests that POEE may be further developed for treating conditions associated with cognitive deficits, especially those linked with cholinergic malfunction.

Mechanism of antiulcerogenic activity of semi-synthetic crotonin obtained from Croton cajucara Benth

The bark of Croton cajucara Benth. is used in Brazilian folk medicine to treat gastrointestinal disorders. Transdehydrocrotonin (DHC) isolated from the bark of Croton cajucara has antiulcerogenic activity25. The presence of similar activity in semi-synthetic crotonin obtained from dehydrocrotonin from Croton cajucara was observed in gastric ulcer-induced models (HCl/ethanol, ethanol, indomethacin, stress and pylorus ligature). The aim of the present study was to assess the mechanisms involved in the antiulcerogenic activity of semi-synthetic crotonin. We investigated the effects of semi-synthetic crotonin on the response to histamine of right atria isolated from guinea pigs and on the response to carbachol of stomach fundus strips from rats. Semi-synthetic crotonin (3, 10 or 30 mM) induced a shift to the right in the concentrationresponse curves to carbachol in the isolated rat stomach at the pD2 level (pD2: 5.42±0.05, 5.76±0.061, 5.77±0.076, 6.48±0.012, respectively), without any alteration in the maximum response. Semi-synthetic crotonin also induced a shift to the right in the concentration-response curves to histamine in guinea pig right atria, pD2 (5.54±0.06, 6.01±0.06, 5.89±0.06, 5.92±0.03) and ( percent) maximum response (80±6.18, 118±6.18, 114±6.18, 122±1.4), respectively. Thus, the protective effect of semi-synthetic crotonin on induced gastric lesions could be due to antagonism of histaminergic and cholinergic effects on gastric secretion.

A program for organizing field research data and bibliography on medicinal plants

In the formulation of working hypotheses based on ethnopharmacological information and publications, a rapid access to data is necessary. We present a microcomputer program especially designed for the organization of field data on traditional uses of medicinal plants (data bank 1) and related bibliographic reference (data bank 2). It was developed in Clipper language and turned into an independent executable through compilation, has many help screens (on-line), allows up to 100,000 entries in each data bank, changing and deletion of data, back-up copies, indexed and not indexed searches, and several possibilities of printing or listing data on the screen. Practical routines for the use of the program are presented, handling the existing 300 field work records and 1,000 references.

Ethnopharmacology and the sustainable development of new plant-derived drugs

This paper presents a reflection based on the experience generated by fifteen years of studies on pharmacology and chemistry of medicinal plants, in a underdeveloped country, Brazil. Although the diversity of the Brazilian flora presents a remarkable opportunity towards the development of new medicinal products, there are only small natural products research groups. Strategies for pharmacological and chemical studies of medicinal plants investigated during this time, major difficulties and ways to bypass them are discussed. The ethnopharmacological approach adopted as a general method by our research team has given good results. Details of our experience in the way to develop new plant durgs are reviewed and updated.

Merging ethnoppharmacology with chemotaxonomy: An approach to unveil bioactive natural products. The case of Psychotria alkaloids as potential analgesics

Psychotria colorata (Will ex R&S) Muell.Arg. flowers are used by "caboclos" in the Amazon as the basis for a homemade analgesic. The study of P. colorata revealed the presence of several pyrrolidinoindoline alkaloids, some with opioid-like analgesic activity in vivo. Neurochemical studies of active alkaloids proved their capability of inhibiting [3H] naloxone binding, confirming the opioid nature of the analgesic activity. These data launched a broader screening of Psychotria species, including P. carthagenensis, P. brachyceras, P. leiocarpa, P. myriantha, P. suterella and P. brachypoda. The analysis of the analgesic activity of several Psychotria species, as well as of specific isolated compounds, allowed a tentative structure/activity relationship and opened a promising research avenue in the search for new analgesic drugs.

Arterite de Takayasu: estudo clínico e angiográfico / Clinical and angiographic features of Takayasu's arteritis

Os autores descrevem os dados clínicos e angiográficos de pacientes com arterite de Takayasu (AT) atendidos no Ambulatório Multidisciplinar de Vasculites da Unifesp/Hospital Säo Paulo entre 1993 e 1997. Foram atendidos 34 pacientes (28 mulheres, 20 de cor branca) com AT. A média de idade por ocasiäo do diagnóstico foi de 27,7 anos e o tempo de doença variou de 2 a 360 meses (mediana de 41 meses). As queixas clínicas por ocasiäo do diagnóstico foram: claudicaçäo de membros superiores em 32 por cento, claudicaçäo de membros inferiores em 23 por cento, emagrecimento em 15 por cento, febre em 15 por cento, insuficiência cardíaca congestiva em 9 por cento, cefaléia em 6 por cento, hipertensäo arterial em 6 por cento e dor precordial em 6 por cento. Durante o seguimento, 60,7 por cento apresentaram ausência ou diminuiçäo de pulso em membro superior, 56 por cento tinham diferença de pressäo arterial > 10 mmHg em membros superiores, 53 por cento tinham sopros arteriais e 29 por cento apresentavam sopros cardíacos. A classificaçäo angiográfica proposta em 1994 para arterite de Takayasu foi aplicada a 28 pacientes com pan-angiografia, que foram classificados como tipo I em 21,4 por cento, tipo IIa em 3,6 por cento, tipo III em 3,6 por cento, tipo IV em 14,3 por cento e tipo V em 57,1 por cento. Nenhum paciente foi classificado como tipo IIb. Três famílias apresentavam mais de um caso de AT: duas famílias com mäe e filha e uma com duas irmäs com AT. Os pacientes com AT atendidos em nosso serviço mostraram grande predomínio da doença em mulheres; 21 por cento dos pacientes apresentavam comprometimento exclusivo dos ramos do arco aórtico (tipo I). Esse padräo de comprometimento é semelhante ao relatado em pacientes japoneses e difere do descrito em pacientes com AT na +ndia