Chemical characterization, antinociceptive and anti-inflammatory effect of Lippia lacunosa, a species used by the Bandeirantes.

ETHNOPHARMACOLOGICAL RELEVANCE: Lippia lacunosa Mart. & Schauer is an endemic plant from the Serra do Espinhaço mountain range located on the Atlantic plateau, Brazil. It is known as "chá de pedestre" and "rosmaninho" in folk medicine. This species has a characteristic mango aroma and is widely used by the population for flu, colds, sinus infections, coughing, relaxing baths, and foot baths after long walks. It is often confused with and, therefore, used interchangeably with L. rotundifolia and L. pseudothea. AIM OF THE STUDY: This study aimed to increase scientific knowledge on the ethnopharmacological use of Lippia lacunosa through the evaluation of the micromolecular composition and anti-inflammatory and antinociceptive activities of the hexane and ethanolic extracts, essential oil, and fractions in mice. MATERIALS AND METHODS: The chemical profile of L. lacunosa extracts and fractions were obtained by chromatographic methods such as Ultra-Performance Liquid Chromatography (UPLC), Gas Chromatography (GC), Column Chromatography (CC), and Thin Layer Chromatography (TLC). Carrageenan-induced paw edema was used to investigate the anti-inflammatory activity in mice. Mechanical allodynia induced by carrageenan and hot plate tests were employed to evaluate the antinociceptive activity. RESULTS: The main constituents found in the essential oil were the monoterpenes myrcene (13.81%), linalool (6.84%), ipsenone (21.2%), and myrcenone (25.44%); and sesquiterpenes elemol (7.30%) and spathulenol (3.15%). The chromatograph fractionation of essential oil yielded a fraction rich in the main compounds (F33), ipsenone and mircenone. In experimental models of paw edema and mechanical allodynia induced by carrageenan (600 µg, 30 µL, i.pl.), the administration of hexane extract, essential oil (50 or 100 mg/kg, p.o.) or majority fraction (10 mg/kg, p.o.) reduced paw edema. The ethanolic extract (100 mg/kg) reduced mechanical allodynia only in the 2 nd h of evaluation. On the other hand, the hexane extract (50 or 100 mg/kg) and essential oil (100 mg/kg), as well as the majority fraction (10 mg/kg), reduced mechanical allodynia throughout the evaluation period. The hexane extract, essential oil, and majority fraction F33 also reduced the heat-induced nociceptive response. Also, majority fraction F33 did not affect the time mice spent in the rota-rod apparatus. CONCLUSIONS: The elucidation of the composition of the essential oil and the demonstration of the activity of L. lacunosa in experimental models of acute inflammation and also in models of nociceptive and inflammatory pain can help to increase knowledge on the ancient ethnopharmacological use by the Bandeirantes, aiming at the evaluation of the species as a candidate for herbal medicine or phytopharmaceutical in the treatment of patients with inflammatory and painful conditions.

The Association of the Inflammatory Index of the Maternal Diet with the Type of Delivery, Maternal Nutritional Status, and the Lipid Profile of Human Milk.

Objective: To evaluate the association of the dietary inflammatory index adjusted for energy (E-DII®) with the evolution of maternal body mass index and human milk lipid (HM) profile in the first 6 months postpartum. Methods: This is a cohort study with 260 Brazilian women (19-43 years old) in the postpartum period. Sociodemographic, gestational, and anthropometric data of the mother were obtained in the immediate postpartum period and in the six monthly meetings. A food frequency questionnaire was applied at baseline and used to calculate the E-DII score. Mature HM was collected, analyzed by gas chromatography coupled with mass spectrometry and the Rose Gottlib method. Generalized estimation equation models were built. Results: Women with >E-DII showed lower adherence to physical activity during pregnancy (p = 0.027), higher frequency of cesarean delivery (p = 0.024), and higher body mass index over time (p < 0.001). At the end of the 6th month, 35 women remained in the study. As for HM, 151 analyses were performed, detecting in the 3rd collection greater fat (%) among those with >E-DII (4.27 ± 1.98 vs. 3.75 ± 2.48; p = 0.003). Conclusion: Diets with >E-DII can impact the type of delivery, change the evolution of maternal nutritional status, and the stability of the lipid profile of HM.

Cinnamon Infusion Reduces Satiety and Increases Energy Intake: A Randomized Crossover Trial.

BACKGROUND/AIMS: This study aimed to evaluate the effects of the intake of a single dose of cinnamon infusion on energy metabolism, appetite responses, and food intake in healthy individuals. METHODS: This was an open randomized crossover clinical trial, with a minimum washout of 7 days, in which two standardized breakfasts were served randomly: one with cinnamon infusion (2 g, Cinnamomum sp.) and one with water (control). The study included 21 healthy volunteers. Energy expenditure (EE), diet-induced thermogenesis (DIT), respiratory quotient (RQ), substrate oxidation, and appetite responses were evaluated, as well as prospective food intake. RESULTS: Ingestion of cinnamon infusion did not alter EE, DIT, RQ, or substrate oxidation. The incremental area under the curve for hunger, satiety, and desire to eat did not differ between the treatment with cinnamon infusion and water; however, there was a reduction in the feeling of satiety (p = 0.021) compared to the control treatment. The energy consumption of the first meal after treatment was higher (p = 0.05) in the treatment with cinnamon infusion than in the control treatment. Regarding macronutrients or food intake throughout the day, there was no difference between treatments. CONCLUSIONS: Although it did not affect energy metabolism, consuming a single dose of cinnamon infusion promoted less feeling of satiety and increased energy intake in the first meal after treatment. The study protocol was registered at the Brazilian Clinical Trials Registry Platform (RBR-5ftg3z).

Euterpe oleracea pulp extract: Chemical analyses, antibiofilm activity against Staphylococcus aureus, cytotoxicity and interference on the activity of antimicrobial drugs.

Euterpe oleracea (Açaí) fruit are widely consumed at the Brazilian Amazon region, and biological potentials such as immunomodulatory and antioxidant have been described for its extracts. However, its antimicrobial properties remain poorly investigated. Here, the antimicrobial and antibiofilm activities of the methanolic extract of an artisanally-manufactured açaí pulp (MEAP) were evaluated against clinical isolates of Staphylococcus aureus. Besides, MEAP interference on the activity of antimicrobial drugs of clinical relevance was explored, and its cytotoxicity against hepatocellular carcinoma cells (HepG2) was investigated. Biochemical and physicochemical properties of the pulp were investigated, and the presence of polyphenols on the extract was confirmed. For the first time, we report that the methanolic extract of açaí pulp is effective against planktonic cells and biofilms of S. aureus, and also decreased the proliferation of HepG2 cells. Statistically significant synergism was observed when the extract was combined to the tested antimicrobials except for erythromycin, and all biochemical and physicochemical parameters ranged within the accepted values established by the Brazilian legislation. Our data open doors for more studies on the antimicrobial activity of phytomolecules isolated from Euterpe oleracea extracts, and also for its combined use with antimicrobial drugs.

Morpho-anatomy and chemical profile of native species used as substitute of quina (Cinchona spp.) in Brazilian traditional medicine. Part II: Remijia ferruginea

ABSTRACT This research is part of a larger study of the Brazilian species that are commonly referred to as "quinas" and used as substitute of Cinchona species. In this study, we have performed the botanical characterization of the stem bark of Remijia ferruginea (A. St.-Hil.) DC., Rubiaceae, by morphological and anatomical description, and the analysis of its chemical profile. Stem bark is thin and has the color and the texture of its external and internal surfaces as diagnostic features. Types and sizes of sclerified cells in the cortical parenchyma and in the secondary phloem are important features for analysis of the transversal sections and in the macerate. Alkaloids, flavonoids and chlorogenic acid were detected in the chemical analysis for TLC. These standard references can be used in the quality control of the bark of quinas.

Medicinal plants recommended by the world health organization: DNA barcode identification associated with chemical analyses guarantees their quality.

Medicinal plants are used throughout the world, and the regulations defining their proper use, such as identification of the correct species and verification of the presence, purity and concentration of the required chemical compounds, are widely recognized. Herbal medicines are made from vegetal drugs, the processed products of medicinal species. These processed materials present a number of challenges in terms of botanical identification, and according to the World Health Organization (WHO), the use of incorrect species is a threat to consumer safety. The samples used in this study consisted of the dried leaves, flowers and roots of 257 samples from 8 distinct species approved by the WHO for the production of medicinal herbs and sold in Brazilian markets. Identification of the samples in this study using DNA barcoding (matK, rbcL and ITS2 regions) revealed that the level of substitutions may be as high as 71%. Using qualitative and quantitative chemical analyses, this study identified situations in which the correct species was being sold, but the chemical compounds were not present. Even more troubling, some samples identified as substitutions using DNA barcoding contained the chemical compounds from the correct species at the minimum required concentration. This last situation may lead to the use of unknown species or species whose safety for human consumption remains unknown. This study concludes that DNA barcoding should be used in a complementary manner for species identification with chemical analyses to detect and quantify the required chemical compounds, thus improving the quality of this class of medicines.

Bitter plants used as substitute of Cinchona spp. (quina) in Brazilian traditional medicine.

ETHNOPHARMACOLOGICAL RELEVANCE: Bitter tasting plant species are used as tonics and have been previously used to treat intermittent fevers in Brazil, the principal symptom of malaria. Many of these species were named quina and were used as substitutes of Cinchona spp., the source of quinine. AIM OF THE STUDY: To present data on these bitter species named quina and to discuss their potential as sources of bioactive substances. MATERIALS AND METHODS: Data about the plants were obtained from a survey of the literature and documents written by early naturalists and clinical doctors living in the 18th and 19th centuries in Brazil. Correlated pharmacological studies were obtained from different scientific databases. RESULTS: A total of 29 species were recorded. The largest number of species belonged to the Rubiaceae family (14), being Remijia ferruginea (A. St.-Hil) DC. the most representative. Strychnos pseudoquina A. St.-Hil. (Loganiaceae), Hortia brasiliana Vand. ex DC. (Rutaceae) and Solanum pseudoquina A. St.-Hil. (Solanaceae) were also frequently mentioned in the historical bibliography. Pharmacological studies have shown the presence of bitter bioactive substances useful to treat digestive disorders and/or with antimalarial activities, in all of the recorded botanic families. CONCLUSION: This study shows that several bitter species named quina were used in the past as substitute of Cinchona spp. and studying these plants can lead to the development of new products.

Morpho-anatomy and chemical profile of native species used as substitutes of quina (Cinchona spp.) in Brazilian traditional medicine. Part I: Polyouratea hexasperma

This research is part of a larger study of the Brazilian species that are commonly referred to as quinas and are usually used as substitute of Cinchona species. The purpose of the present study was the botanical characterization of the whole and powdered stem bark of Polyouratea hexasperma (A. St.-Hil.) Tiegh., Ochnaceae, by morphological and anatomical description, and the analysis of its chemical profile. The external texture of the bark, the whitened pit in the inner bark and the presence of cristarque cells, as well as the shape and arrangement of other lignified cells, are the most important macroscopic and microscopic features for the characterization of the bark. Chlorogenic and cafeic acids were detected in the chemical analysis and can also be used in the identification of the bark.

Changes in the trade in native medicinal plants in Brazilian public markets.

Plants continue to be an important source of new bioactive substances. Brazil is one of the world's mega-diverse countries, with 20 % of the world's flora. However, the accelerated destruction of botanically rich ecosystems has contributed to a gradual loss of native medicinal species. In previous study, we have observed a fast and intensive change in trade of medicinal plants in an area of Amazon, where human occupation took place. In this study, we surveyed 15 public markets in different parts of Brazil in search of samples of 40 plants used in traditional medicine and present in first edition of Brazilian Official Pharmacopoeia (FBRAS), published in 1926. Samples of plants commercialized as the same vernacular name as in Pharmacopoeia were acquired and submitted to analysis for authentication. A total of 252 plant samples were purchased, but the laboratory analyses showed that only one-half of the samples (126, 50.2 %) were confirmed as the same plant species so named in FBRAS. The high number of unauthenticated samples demonstrates a loss of knowledge of the original native species. The proximity of the market from areas in which the plant occurs does not guarantee that trade of false samples occurs. The impact of the commerce of the substitute species on their conservation and in public health is worrying. Strategies are necessary to promote the better use and conservation of this rich heritage offered by Brazilian biodiversity.

Nephroprotective Effect of Echinodorus macrophyllus Micheli on Gentamicin-Induced Nephrotoxicity in Rats.

BACKGROUND/AIMS: Leaves of Echinodorus macrophyllus (EM), from the Alismataceae family, have been used in Brazilian folk medicine for their anti-inflammatory and diuretic properties. In this work, the diuretic and nephroprotective activities of crude extracts of EM were evaluated. METHODS: Normal Wistar rats were given 0.9% NaCl containing either EM (10-300 mg/kg), furosemide (13 mg/kg) or arginine vasopressin (0.2 mg/kg). Thereafter, the rats were individually housed in metabolic cages, and urine volume was measured every 30 min for a total of 3 h. Acute kidney injury was induced by gentamicin (GM, 80 mg·kg(-1)·day(-1), b.i.d., 5 days). Along with GM, 0.9% NaCl (control) or EM (30 mg/kg) was given to the rats by gavage. RESULTS: EM produced a dose-dependent reduction in urine elimination. EM was effective in reversing all GM-induced alterations such as polyuria and glomerular filtration rate reduction. The GM-induced morphological alterations were not observed when EM was given concomitantly with GM. CONCLUSION: This study provides evidence that EM possesses nephroprotective effect which indicates that EM may have therapeutic applications in GM-induced acute kidney injury.

Influence of the preparation method on the low efficacy of multi-herb commercial products: the example of João da Costa e Associações / Influência do método de preparação na baixa eficácia de produtos fitoterápicos: o exemplo do João da Costa e Associações

Native medicinal plants have been used for decades by Brazilian pharmaceutical companies to create commercial products. In this study, we have investigated the herb-combined product João da Costa e Associações® (JCA) commercialized for thirty years to treat dysmenorrhoea. JCA is prepared by decoction of Himatanthus lancifolius (Muell. Arg.) Woodson (Apocynaceae), Chondodendron platyphyllum Miers (Menispermaceae), Gossypium herbaceum L. (Malvaceae), Rosmarinus officinalis L. (Lamiaceae) and Echites peltata (Apocynaceae), followed by addition of sugar. The efficacy of JCA was verified by antinociceptive studies. The chemical composition was determined by fingerprint analysis in HPLC/ DAD. A weak inhibition of the second phase of the nociceptive effect induced by formalin indicated an activity similar to those steroids and not-steroids anti-inflammatories. Despite being prepared by decoction of five plants, the fingerprint analysis showed only two peaks. None of them corresponds to the chemical compounds observed in ethanol extracts prepared with the same plant material. We argue that the methods of preparation of the formulas should be considered in studies of multi-herbs products, since they can be the responsible for inefficacy or low activity of such products.

Plantas medicinais nativas do Brasil foram usadas por décadas pelas indústrias farmacêuticas nacionais para criar seus produtos. Neste estudo, foi investigado o produto João da Costa e Associações® (JCA) comercializado por mais de trinta anos para o tratamento de dismenorréia e outros problemas relacionados à saúde da mulher. JCA é preparado pela decocção de Himatanthus lancifolius (Muell. Arg.) Woodson (Apocynaceae), Chondodendron platyphyllum Miers (Menispermaceae), Gossypium herbaceum L. (Malvaceae), Rosmarinus officinalis L. (Lamiaceae) e Echites peltata (Apocynaceae), seguido de adição de açúcar. A eficácia de JCA foi verificada por meio da avaliação da atividade antinociceptiva. Já a composição química foi determinada por analises em HPLC/ DAD. Uma fraca inibição da segunda fase da nocicepção foi observada no teste da formalina, indicando uma ação semelhante aos antiinflamatórios esteroidais e não esteroidais. Apesar de ser preparado pela decocção de cinco plantas, a análise no HPLC apresentou somente dois picos, e nenhum deles correspondeu aos componentes observados nos extratos etanólicos preparados com as mesmas plantas. Os resultados sugerem que o método de preparação de JCA promove a perda dos componentes químicos das plantas e interfere consideravelmente na eficácia do produto.

Influence of Brazilian herbal regulations on the use and conservation of native medicinal plants.

For decades, several native medicinal species have been used by the pharmaceutical industry in Brazil to create commercial products. In 1995, the Ministry of Health, following the recommendations of the World Health Organization, established herbal regulations (RDC 17) in order to improve the quality of such products. In fact, only plant species with conclusive results from pharmacological and toxicological studies can be used for creating medicines. In this study, we evaluated the consequences of RDC 17 on the use and conservation of native medicinal plants by comparing the plant material used by six companies in 1995/1996 and 10 years later (2005/2006). Eighty-three different species were used in 1995/1996, 50 of them native (60.2%), 16 exotic (19.3%), and 17 imported (20.5%). In 2005/2006, 44 species were used by the companies and only 19 (43.2%) were native. The category of plant material that saw the largest decrease in use was roots, and in 2005/2006 leaves were more used. The study shows a strong reduction in the collection of native species signalizing the importance of herbal regulations on their conservation. It also points to the need for pharmacological and toxicological studies of the Brazilian native medicinal flora, as well as studies on their ecology and conservation.

Traditional uses of American plant species from the 1st edition of Brazilian Official Pharmacopoeia / Usos tradicionais das plantas americanas presentes na 1ª Edição da Farmacopéia Brasileira

The first edition of the Brazilian Official Pharmacopoeia (FBRAS), published in 1929, is a rich source of information about American medicinal plants, since it lists species used in both traditional and conventional medicine. In this study, we have performed a survey of the traditional uses of plants described in eighty-seven Monographs from the FBRAS in twenty bibliographies written from the 19th century to the 1970s. Eighty-six different traditional uses are described in three or more books; some of them were cited in ten or more books, illustrating their widespread use and importance in medicine. The species from the first edition of the FBRAS have a long tradition of medical utility, which is confirmed by historical records. In surveying these medically relevant species, we hope to encourage policy makers and the scientific public as a whole to engage in a strong debate in an attempt to improve and facilitate the pharmacological study of these species.

A 1ª Edição da Farmacopéia Brasileira, publicada em 1929, é uma rica fonte de informação sobre as plantas medicinais nativas das Américas, uma vez que ela lista as espécies usadas na medicina tradicional e oficial da época. Neste estudo, foi feita uma extensa revisão sobre os usos tradicionais de plantas descritas em oitenta e sete Monografias presentes na FBRAS 1ª Edição, em vinte livros publicados do século 19 até a década de 1970. Oitenta e seis diferentes usos medicinais foram citados e três ou mais livros; mesmos usos foram descritos em dez ou mais referências, mostrando seu amplo emprego e importância como medicamento. Os resultados mostram que as espécies nativas presentes na 1ª Edição da FBRAS têm longa tradição em uso, confirmado pelos dados históricos. É necessário, portanto, que estudos farmacológicos com essas plantas sejam estimulados, como forma de promover seu melhor aproveitamento.

Effect of triterpene saponins from roots of Ampelozizyphus amazonicus Ducke on diuresis in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Ampelozizyphus amazonicus Ducke is a plant used in Brazilian folk medicine to both prevent malaria and act as a depurative. AIM OF THE STUDY: We have investigated the effects of an ethanol crude extract of roots of Ampelozizyphus amazonicus (CEAaD), a chemically characterized saponin mixture (SAPAaD), as well as a saponin-free fraction (SAPAaD-free) obtained from CEAaD on diuresis in rats. MATERIALS AND METHODS: Wistar rats under ad libitum water conditions or water deprivation for 12h prior to the start of the experiment were volume-expanded with 0.9% NaCl (4% body weight, by gavage) containing either CEAaD, SAPAaD, or SAPAaD-free at the doses indicated in the text. Rats were individually housed in metabolic cages, and urine volume was measured every 30 min throughout the experiment (3 h). RESULTS: CEAaD increased urine volume in rats under conditions of both free access to water and under water deprivation. In the latter condition, CEAaD (150 mg/kg) increased the urine volume from zero to 0.9+/-0.1 ml/120 min, n=6). Similarly, the SAPAaD-free (50-200 mg/kg) mixture also increased the urine volume. In contrast, SAPAaD (12.5-1000 mg/kg) produced a significant reduction (p<0.01) in diuresis under conditions of both water deprivation and with free access to water prior to the start of the experiment. CONCLUSION: Our data indicate that CEAaD contains compounds that cause both diuresis and antidiuresis and that the antidiuretic effect is due mainly to the presence of saponins.

Medicinal plants and other botanical products from the Brazilian Official Pharmacopoeia / Plantas medicinais e outros produtos vegetais da Farmacopéia Brasileira

In this paper, we describe the results of a thorough survey in the four editions of the Brazilian Official Pharmacopoeia (FBRAS), in a search for data about the plants and other botanical products included in them. The growth of the pharmaceutical industry since the second half of last century markedly affected the Brazilian official medicine. The paper analyses the transformation within the Pharmacopoeia, focusing on the presence of Monographs for Native medicinal plants. The result showed a strong substitution of Native American medicinal plants by industrialized medicine and foreign medicinal plants in FBRAS and confirms the necessity of investiments in research for the validation of Brazilian Native plants.

Este artigo descreve os resultados de uma extensa revisão efetuada nas quatro edições da Farmacopéia Brasileira, buscando dados sobre as plantas medicinais e outros produtos vegetais nela descritos. O crescimento da indústria farmacêutica internacional a partir da segunda metade do século passado afetou intensamente a medicina oficial no Brasil. O texto faz uma breve análise das transformações sofridas pela Farmacopéia, tomando como base a existência de Monografias para as espécies vegetais nativas. Os resultados do estudo demonstram uma intensa substituição das plantas nativas do Brasil por medicamentos industrializados e outros produtos vegetais estrangeiros, confirmando assim a necessidade de investimentos em pesquisa de validação das nossas plantas medicinais.