NMR and multivariate methods: Identification of chemical markers in extracts of pedra-ume-caá and their antiglycation, antioxidant, and enzymatic inhibition activities.

INTRODUCTION: In Brazil, the plant group popularly known as "pedra-ume-caá" is used in folk medicine for the treatment of diabetes, and its raw material is commonly sold. OBJECTIVE: The aim of the study was to apply a method for chemical identification of extracts of dry pedra-ume-caá leaves using HPLC-high-resolution mass spectrometry (HRMS) and NMR and develop a multivariate model with NMR data to authenticate commercial samples. In addition, to evaluate the biological activities of the extracts. MATERIALS AND METHODS: Dry extracts of Myrcia multiflora, Myrcia amazonica, Myrcia guianensis, Myrcia sylvatica, Eugenia punicifolia leaves, and 15 commercial samples (sold in Manaus and Belém, Brazil) were prepared by infusion. All the extracts were analysed using HPLC-high-resolution mass spectrometry (HRMS), NMR, principal component analysis (PCA), and hierarchical cluster analysis (HCA). The antidiabetic effect of extracts was evaluated according to enzymatic inhibition. Their content of total phenols, cell viability, and antioxidant and antiglycation activities were also determined. RESULTS: HPLC-HRMS and NMR analysis of these extracts permitted the identification of 17 compounds. 1H NMR data combined with multivariate analyses allowed us to conclude that catechin, myricitrin, quercitrin, and gallic and quinic acids are the main chemical markers of pedra-ume-caá species. These markers were identified in 15 commercial samples of pedra-ume-caá. Additionally, only the extracts of M. multiflora and E. punicifolia inhibited α-glucosidase. All the extracts inhibited the formation of advanced glycation end products (AGEs) and showed free-radical-scavenging activity. These extracts did not present cytotoxicity. CONCLUSION: This study revealed the chemical markers of matrices, and it was possible to differentiate the materials marketed as pedra-ume-caá. Moreover, this study corroborates the potential of these species for treating diabetes.

Hypoglycemic effect of the dry leaf extract of Myrcia multiflora in streptozotocin-induced diabetic mice.

ETHNOPHARMACOLOGICAL RELEVANCE: Myrcia multiflora (Lam) DC. is a medicinal plant used in folk medicine for diabetes control, mainly in the Brazilian Amazon. The leaves of this species has already demonstrated antidiabetic properties; however, in mice with type 2 diabetes (DM2), the cumulative effect of the consumption of the dry extract of M. multiflora leaves (Mm) has not yet been reported. AIM OF THE STUDY: To investigate the effect of the dry extract obtained from the infusion of the dried leaves of M. multiflora on the blood glucose levels of diabetic mice. MATERIALS AND METHODS: DM2 was induced in Swiss male mice by a single intraperitoneal injection of streptozotocin [150 mg/kg body weight (bw)]. The animals were divided into two control groups (healthy and diabetic without treatment) and three sample groups that received Mm (25 and 50 mg/kg bw) and acarbose (200 mg/kg bw) by gavage once daily for 28 days (D28). Additionally, biochemical parameters, thiobarbituric acid reactive species (TBARS) levels in the liver, and histopathological analyses of the kidneys and liver were performed. RESULTS: On the seventh day of treatment, a 74.7% reduction in glucose levels were observed in the group of diabetic animals treated with Mm (50 mg/kg bw) when compared to the beginning of the treatment. At D28, the hypoglycemic effect was maintained. The results of the biochemical and histopathological parameters and the TBARS levels suggest that this dry extract exerts nephro- and hepatoprotective effects. CONCLUSIONS: The findings demonstrate the potential that this extract has to inhibit the α-glucosidase enzyme, and it acts similarly to the positive control acarbose. Furthermore, this extract is nephro- and hepatoprotective. Therefore, this dry extract has the potential to be an adjuvant for DM2, which corroborates its use in folk medicine.

Hypoglycemic effect and toxicity of the dry extract of Eugenia biflora (L.) DC. leaves.

ETHNOPHARMACOLOGICAL RELEVANCE: The leaves of Eugenia biflora (Myrtaceae) are traditionally used by Amazonian populations for the control of diabetes. However, their chemical composition has not yet been described and pharmacological evidence has not been reported. OBJECTIVE: This study aimed to identify the chemical constituents and evaluate the hypoglycemic and toxic effect of the dry extract of the E. biflora leaves (DEEB). MATERIALS AND METHODS: DEEB, obtained by infusion, was analyzed using LC-HRMS and NMR, whose the catechin flavonoid was quantified using NMR. The antidiabetic effect of DEEB was evaluated according to its inhibition of the enzymes α-amylase and α-glucosidase, as well as the content of total phenols, free radical scavengingand antiglycation activities, and its in vitro cell viability. Oral maltose tolerance and chronic multiple dose tests (28 days) in streptozotocin-induced diabetic mice (STZ) were performed. The hypoglycemic effect and toxicity of this extract were evaluated in the multiple dose assay. Biochemical parameters, hemolysis, and levels of the thiobarbituric acid reactive species in the liver were investigated and histopathological analyses of the kidneys and liver were performed. RESULTS: Eight phenolic compounds were identified, with catechin (15.5 ± 1.7 mg g-1) being the majority compound and a possible chemical marker of DEEB. The extract showed inhibition activity of the enzyme α-glucosidase. Chronic administration of DEEB (50 mg/kg of body weight) reduced glucose levels in diabetic animals, similar to acarbose; however, DEEB (100 and 200 mg/kg bw) caused premature death of mice by D22 of the treatment. Our data indicate that one of the mechanisms of toxicity in DEEB may be related to the aggravation of oxidative stress in the liver. This histopathological study indicated that DEEB failed to minimize the progression of the toxicity of diabetes caused by STZ. CONCLUSIONS: This study demonstrated the hypoglycemic potential of E. biflora leaves. However, the prolonged use of this tea can be harmful to its users due to its considerable toxicity, which needs to be better investigated.

Libidibia ferrea (jucá) anti-inflammatory action: A systematic review of in vivo and in vitro studies.

Libidibia ferrea (Mart. ex Tul.) L. P. Queiroz (jucá) is a plant extensively used in the Brazilian folk medicine for the treatment of the inflammatory process. Primary studies have focused on the verification of these biological activities, highlighting the role of this plant in inflammatory conditions. This systematic review aimed to critically establish which part of the plant and what type of plant extract present the highest evidence of anti-inflammatory activity as in vivo and in vitro experimental models. This study has followed the recommendations by PRISMA and was registered in the PROSPERO database under number CRD42020159934. The literature review was carried out in several medical and scientific databases (Google Scholar, LILACS, ProQuest, PubMed, ScienceDirect, Scopus and Web of Science) in studies published up to February 2020 and updated on March 2021. No language restriction was made to this search. Eligibility criteria were adopted instead. The risk of bias was evaluated through SYRCLE's RoB tool for the in vivo studies. 609 studies were initiated to identify the whole and the subsequent steps of screening. 13 studies remained in the results (10 in vivo and 3 in vitro). In most studies the risk of bias was low or unclear. The high risk of bias was related to the risk of attrition and reporting bias. The fruit and the aqueous extract were identified as the most used in the studies carried out on the qualitative analysis and the results of the in vivo and in vitro studies were conducive to the anti-inflammatory action, a meta-analysis could not be performed due to heterogeneity between studies and the potential risk of bias to estimate the side effects. Therefore, the implementation of in vivo studies following the international guidelines could collaborate with analyses of the anti-inflammatory effect of jucá.

qNMR quantification of phenolic compounds in dry extract of Myrcia multiflora leaves and its antioxidant, anti-AGE, and enzymatic inhibition activities.

Myrcia multiflora (Lam.) DC. is often used in Brazilian folk medicine to control diabetes. Analysis using HPLC-HRMS and NMR of the dry extract from the infusion of leaves of this species revealed twelve phenolic compounds. Among these compounds, chlorogenic acid (1), 4-O-caffeoylquinic acid (2), corilagin (3), chebulagic acid (4), pedunculagin (5), quercetin-3-O-ß-2″-galloylglucoside (7), and kaempferol-3-O-rhamnoside (12) are described for the first time in this matrix. Furthermore, six compounds were quantified using qNMR. The compounds in the dry extracts are 3, 6 (myricetin-3-O-d-glucoside), 8 (myricitrin), 9 (hyperoside), 10 (guaijaverin) and 11 (quercitrin). These compounds may be considered chemical markers in this matrix. In addition, this extract presents activities of α-glucosidase inhibition (IC50 = 79.9 µg mL-1) and glycation in vitro (IC50 = 10.2 µg mL-1), in addition to antioxidant activity against DPPH and ABTS radicals (1,856.7 and 1,032.0 µmol TEq, respectively). This extract did not show significant cytotoxicity in human fibroblasts. Therefore, the enzymatic inhibition, anti-AGE (advanced glycation end-products) and antioxidant activities of Myrcia multiflora leaves corroborated its antidiabetic therapeutic potential and instigates future preclinical studies aimed at the treatment of diabetes mellitus and its complications.

Essential Oil from Bark of Aniba parviflora (Meisn.) Mez (Lauraceae) Reduces HepG2 Cell Proliferation and Inhibits Tumor Development in a Xenograft Model.

Aniba parviflora (Meisn.) Mez (Lauraceae) is an aromatic plant of the Amazon rainforest, which has a tremendous commercial value in the perfumery industry; it is popularly used as flavoring sachets and aromatic baths. In Brazilian folk medicine, A. parviflora is used to treat victims of snakebites. Herein, we analyzed the chemical composition of A. parviflora bark essential oil (EO) and its effect on the growth of human hepatocellular carcinoma HepG2 cells in vitro and in vivo. EO was obtained by hydrodistillation and characterized by GC-MS and GC-FID. The main constituents of EO were linalool (16.3±3.15), α-humulene (14.5±2.41 %), δ-cadinene (10.2±1.09 %), α-copaene (9.51±1.12 %) and germacrene B (7.58±2.15 %). Initially, EO's cytotoxic effect was evaluated against five cancer cell lines (HepG2, MCF-7, HCT116, HL-60 and B16-F10) and one non-cancerous one (MRC-5), using the Alamar blue method after 72 h of treatment. The calculated IC50 values were 9.05, 22.04, >50, 15.36, 17.57, and 30.46 µg/mL, respectively. The best selectivity was for HepG2 cells with a selective index of 3.4. DNA Fragmentation and cell cycle distribution were quantified in HepG2 cells by flow cytometry after a treatment period of 24 and 48 h. The effect of EO on tumor development in vivo was evaluated in a xenograft model using C.B-17 SCID mice engrafted with HepG2 cells. In vivo tumor growth inhibition of HepG2 xenograft at the doses of 40 and 80 mg/kg were 12.1 and 62.4 %, respectively.

Pedra-ume caá fruit: An Amazon cherry rich in phenolic compounds with antiglycant and antioxidant properties.

Species of Eugenia have been used as an antidiabetic natural source. Chemical, antioxidant and antiglycant screening of extracts from pedra-ume caá (Eugenia punicifolia) fruits were performed. 1H NMR assisted by non-supervised chemometric methods were employed for the evaluation of the chemical profiles which were distinguished according to the color of fruit maturation stages, as well as for pulp and seed fruit. Furthermore, 1H NMR fingerprint analysis of the crude extract allowed the identification of quercitrin and myricitrin, beside other nine compounds. The extracts of the yellow (YP) and green (GP) pulps presented higher antiglycant and antioxidant activities. Fresh juice from E. punicifolia was encapsulated in microcapsules produced with dextrose equivalent (DE) of 10, 20 or 30 as wall materials for the maintainment of their antioxidant and antiglycant properties. The more efficient retention of the bioactive compounds was found using the DE30. The Encapsulation Efficiency (EE) and the Retention Efficiency (RE) of this system was found around 89.7% and 97.6%, respectively. In addition, NMR spectra revealed the presence of flavonoids O-glycosylated (quercitrin and myricitrin) which might be related to the antiglycant and antioxidant activities. The YP presented larger content of quercitrin (117.6 ±â€¯0.4 mg per each 100 g of fresh fruit). Therefore, pedra-ume caá should be employed as an alternative nutraceutical source, as well as intherapeutic pourposes.

Remela de cachorro (Clavija lancifolia Desf.) fruits from South Amazon: Phenolic composition, biological potential, and aroma analysis.

Remela de cachorro (Clavija lancifolia Desf.) is an Amazonian native fruit consumed specially in the Purus microregion. Because of its rarity, restricted consumption, and the lack of knowledge about its chemical composition, remela de cachorro fruit was studied in relation to its phenolic and aroma constitution. Using liquid chromatography tandem mass spectrometry (LC-MS/MS), 11 compounds (flavonoids and its glucosides along with organic acids) were tentatively identified by fragmentation patterns. A previously validated method was applied to quantify common antioxidant compounds in the raw pulps, for which kaempferol was the main compound. Gas chromatography mass spectrometry (GC-MS) with headspace solid-phase microextraction (HS-SPME) was employed to assess the aroma composition of remela de cachorro fruit. A total of 27 volatile organic compounds (VOCs) were identified for this fruit, for which benzaldehyde and linalool were the main VOCs. Furthermore, biological activities, such as antioxidant capacity (ABTS, DPPH, and ORAC methods), cytotoxicity, and α-glucosidase and lipase inhibitions of the hydroalcoholic extract of remela de cachorro fruit were evaluated. In vitro biological assays revealed the potential of this fruit as a bioactive food that should be further studied and explored in Amazonian products.

Antioxidant activity and peroxidase inhibition of Amazonian plants extracts traditionally used as anti-inflammatory.

BACKGROUND: The Amazon is the largest rainforest in the world and is home to a rich biodiversity of medicinal plants. Several of these plants are used by the local population for the treatment of diseases, many of those with probable anti-inflammatory effect. The aim of the present investigation was to evaluate the in vitro antioxidant and anti-peroxidases potential of the ethanol extracts of five plants from the Brazilian Amazon (Byrsonima japurensis, Calycophyllum spruceanum, Maytenus guyanensis, Passiflora nitida and Ptychopetalum olacoides). METHODS: DPPH, ABTS, superoxide anion radical, singlet oxygen and the ß-carotene bleaching methods were employed for characterization of free radical scavenging activity. Also, total polyphenols were determined. Antioxidant activities were evaluated using murine fibroblast NIH3T3 cell. Inhibition of HRP and MPO were evaluated using amplex red® as susbtract. RESULTS: The stem bark extracts of C. spruceanum and M. guyanensis provided the highest free radical scavenging activities. C. spruceanum exhibited IC50 = 7.5 ± 0.9, 5.0 ± 0.1, 18.2 ± 3.0 and 92.4 ± 24.8 µg/mL for DPPH(•), ABTS(+•), O2 (-•) and (1)O2 assays, respectively. P. olacoides and C. spruceanum extracts also inhibited free radicals formation in the cell-based assay. At a concentration of 100 µg/mL, the extracts of C. spruceanum, B. japurensis inhibited horseradish peroxidase by 62 and 50 %, respectively. C. spruceanum, M. guyanensis, B. japurensis also inhibited myeloperoxidase in 72, 67 and 56 %, respectively. CONCLUSIONS: This work supports the folk use these species that inhibited peroxidases and exhibited significant free radical scavenging and antioxidant activities what can be related to treatment of inflammation.

In vitro and in vivo anti-malarial activity of plants from the Brazilian Amazon.

BACKGROUND: The anti-malarials quinine and artemisinin were isolated from traditionally used plants (Cinchona spp. and Artemisia annua, respectively). The synthetic quinoline anti-malarials (e.g. chloroquine) and semi-synthetic artemisinin derivatives (e.g. artesunate) were developed based on these natural products. Malaria is endemic to the Amazon region where Plasmodium falciparum and Plasmodium vivax drug-resistance is of concern. There is an urgent need for new anti-malarials. Traditionally used Amazonian plants may provide new treatments for drug-resistant P. vivax and P. falciparum. Herein, the in vitro and in vivo antiplasmodial activity and cytotoxicity of medicinal plant extracts were investigated. METHODS: Sixty-nine extracts from 11 plant species were prepared and screened for in vitro activity against P. falciparum K1 strain and for cytotoxicity against human fibroblasts and two melanoma cell lines. Median inhibitory concentrations (IC50) were established against chloroquine-resistant P. falciparum W2 clone using monoclonal anti-HRPII (histidine-rich protein II) antibodies in an enzyme-linked immunosorbent assay. Extracts were evaluated for toxicity against murine macrophages (IC50) and selectivity indices (SI) were determined. Three extracts were also evaluated orally in Plasmodium berghei-infected mice. RESULTS: High in vitro antiplasmodial activity (IC50 = 6.4-9.9 µg/mL) was observed for Andropogon leucostachyus aerial part methanol extracts, Croton cajucara red variety leaf chloroform extracts, Miconia nervosa leaf methanol extracts, and Xylopia amazonica leaf chloroform and branch ethanol extracts. Paullinia cupana branch chloroform extracts and Croton cajucara red variety leaf ethanol extracts were toxic to fibroblasts and or melanoma cells. Xylopia amazonica branch ethanol extracts and Zanthoxylum djalma-batistae branch chloroform extracts were toxic to macrophages (IC50 = 6.9 and 24.7 µg/mL, respectively). Andropogon leucostachyus extracts were the most selective (SI >28.2) and the most active in vivo (at doses of 250 mg/kg, 71% suppression of P. berghei parasitaemia versus untreated controls). CONCLUSIONS: Ethnobotanical or ethnopharmacological reports describe the anti-malarial use of these plants or the antiplasmodial activity of congeneric species. No antiplasmodial activity has been demonstrated previously for the extracts of these plants. Seven plants exhibit in vivo and or in vitro anti-malarial potential. Future work should aim to discover the anti-malarial substances present.

Anti-inflammatory action of Justicia acuminatissima leaves

AbstractIn Amazonas State (Brazil), Justicia acuminatissima (Miq.) Bremek., Acanthaceae, leaf teas are used in folk medicine to treat several inflammatory illnesses. In order to validate this medicinal application, we analyzed the acute toxicity and antioxidant, antiedematogenic and antinociceptive potentials of an aqueous extract of this species, using culture cells and animal models. The aqueous extract did not cause toxic effects on human lymphocytes in high concentration (400 μg/ml), neither on mice treated with high doses (5000 mg/kg) in an acute toxicity analysis by oral route, and also did not cause lesions in the gastric mucosa of animals treated with 300 mg/kg, which was the maximal dose used in the anti-inflammatory screening. The aqueous extract caused inhibition of inflammatory pain in formalin-induced paw licking test with all tested doses, 30, 100 and 300 mg/kg, and antiedematogenic activity at 100 and 300 mg/kg. Additionally, the aqueous extract presented statistically significant action on the release of nitric oxide by lipopolysaccharide-activated macrophages. These results and other preliminary studies support the folk use of this species, and further investigation of its action mechanism by inhibition of COX-2 or related metabolite would be interesting.

Biological activities and cytotoxicity of diterpenes from Copaifera spp. Oleoresins.

Copaifera spp. are Amazonian species widely studied and whose oleoresins are used by local people for various medicinal purposes. However, a detailed study of the activity of the main phytochemical components of these oleoresins remains to be done. Here, we studied the cytotoxicity and in vitro anti-inflammatory effects of six diterpene acids: copalic, 3-hydroxy-copalic, 3-acetoxy-copalic, hardwickiic, kolavic-15-metyl ester, and kaurenoic, isolated from the oleoresins of Copaifera spp. The diterpenes did not show cytotoxicity in normal cell lines, nor did they show significant changes in viability of tumoral line cells. The 3-hydroxy-copalic was able to inhibit the enzyme tyrosinase (64% ± 1.5%) at 250 µM. The kolavic-15-metyl ester at 200 µM showed high inhibitory effect on lipoxygenase (89.5% ± 1.2%). Among the diterpenes tested, only kaurenoic and copalic acids showed significant hemolytic activities with 61.7% and 38.4% at 100 µM, respectively. In addition, it was observed that only the copalic acid (98.5% ± 1.3%) and hardwickiic acid (92.7% ± 4.9%) at 100 mM inhibited nitric oxide production in macrophages activated by lipopolysaccharide. In this assay, the diterpenes did not inhibit tumor necrosis factor-α production. The acids inhibited the production of IL-6, 3-acetoxy-copalic (23.8% ± 8.2%), kaurenoic (11.2% ± 5.7%), kolavic-15-methyl ester (17.3% ± 4.2%), and copalic (4.2% ± 1.8%), respectively, at 25 µM. The kaurenoic, 3-acetoxy-copalic and copalic acids increased IL-10 production. This study may provide a basis for future studies on the therapeutic role of diterpenic acids in treating acute injuries such as inflammation or skin disorders.

Anti-Inflammatory Activity of Triterpenes Isolated from Protium paniculatum Oil-Resins.

Protium is the main genus of the Burseraceae family and one of the most common genera in South America, with an important species called "breu." Gum and oil-resins of this species are used as tonic and stimulant and for the treatment of ulcers and inflammation. The present study aims to isolate and investigate the anti-inflammatory activity of triterpene compounds isolated from oil-resin of Protium paniculatum. The pentacyclic triterpenes α,ß-amyrin, acetylated α,ß-amyrin, α,ß-amyrone, and brein/maniladiol did not alter the viability of murine J774 macrophages (IC50 > 20 µg/mL), with the exception of mixture of brein/maniladiol which showed moderate cytotoxic activity. Also it was observed that compounds at 10 µg/mL inhibited more than 80% of production of NO(•), although only α,ß-amyrin was able to inhibit the production of TNF-α (52.03 ± 2.4%). The compounds inhibited the production of IL-6 and induced the production of IL-10 in murine J774 macrophages stimulated by LPS. α,ß-Amyrone inhibited the expression of COX-2 and also inhibited the formation of paw or ear edema in rats and mice, having a quick and immediate effect. This study may provide the basis for future investigations on the therapeutic role of α,ß-amyrone in treating inflammation.

In vitro and in vivo anti-malarial activity of limonoids isolated from the residual seed biomass from Carapa guianensis (andiroba) oil production.

BACKGROUND: Carapa guianensis is a cultivable tree used by traditional health practitioners in the Amazon region to treat several diseases and particularly symptoms related to malaria. Abundant residual pressed seed material (RPSM) results as a by-product of carapa or andiroba oil production. The objective of this study was to evaluate the in vitro and in vivo anti-malarial activity and cytotoxicity of limonoids isolated from C. guaianensis RPSM. METHODS: 6α-acetoxyepoxyazadiradione (1), andirobin (2), 6α-acetoxygedunin (3) and 7-deacetoxy-7-oxogedunin (4) (all isolated from RPSM using extraction and chromatography techniques) and 6α-hydroxy-deacetylgedunin (5) (prepared from 3) were evaluated using the micro test on the multi-drug-resistant Plasmodium falciparum K1 strain. The efficacy of limonoids 3 and 4 was then evaluated orally and subcutaneously in BALB/c mice infected with chloroquine-sensitive Plasmodium berghei NK65 strain in the 4-day suppressive test. RESULTS: In vitro, limonoids 1-5 exhibited median inhibition concentrations (IC50) of 20.7-5.0 µM, respectively. In general, these limonoids were not toxic to normal cells (MRC-5 human fibroblasts). In vivo, 3 was more active than 4. At oral doses of 50 and 100 mg/kg/day, 3 suppressed parasitaemia versus untreated controls by 40 and 66%, respectively, evidencing a clear dose-response. CONCLUSION: 6α-acetoxygedunin is an abundant natural product present in C. guianensis residual seed materials that exhibits significant in vivo anti-malarial properties.

Effects of Passiflora nitida Kunth leaf extract on digestive enzymes and high caloric diet in rats.

The present study investigated inhibition of pancreatic lipase and metabolic effects of high caloric diet in rats. The Passiflora nitida hydroethanol leaf extract (PNE) was used in in vitro assays or administered to rats to study dyslipidemia. Inhibition of lipase in vitro was studied by a spectrophotometric assay using orlistat as the positive control. The effects of PNE on reduction of postprandial triglyceride were studied by oral fat-overloading in rats. Metabolic alterations were induced using the cafeteria diet and 4 weeks post-treatment with PNE or orlistat and blood samples were collected and biochemical analyses were performed. Liver and retroperitoneal fat tissues were obtained to analyze weight and steatosis. IC50 (lg/mL) values for pancreatic lipase inhibition were 21.2 ± 0.8 and 0.1 ± 0.01 for PNE and orlistat, respectively. Oral administration of lipid emulsion resulted in postprandial hypertriglyceridemia at 3 h postadministration and when rats were then administered PNE and orlistat there was decreased of triglyceride levels by 15 % compared to control. Although the energy consumption by the cafeteria diet had been higher, there was no significant weight gain observed in the study groups. The cafeteria diet resulted in a significant increase of weight in the retroperitoneal fat and hypertriglyceridemia levels that could be significantly reduced by PNE and orlistat treatment. We hypothesized that PNE administration prevented the hypertriglyceridemia in rats with a high caloric diet, possibly owing to reduction of lipid absorption and pancreatic lipase inhibition.

Effects of diet supplementation with Camu-camu (Myrciaria dubia HBK McVaugh) fruit in a rat model of diet-induced obesity.

Amazonian Camu-camu fruit (Myrciaria dubia HBK Mc Vaugh) has attracted interest from food and cosmetics industries because of its rich content of vitamin C, flavonoids and anthocyanins. The goal of this study was investigates the antiobesity action of the ingestion of the Camu-camu pulp in a rat model of diet-induced obesity. Wistar rats with obesity induced by subcutaneous injection of monosodium glutamate receiving diet ad libitum. The rats were divided in two groups: an experimental group that ingested 25 mL/day of Camu-camu pulp (CCG) and a non treated group (CG). After 12 weeks, the animals were sacrificed. Blood, liver, heart, white adipose tissues were collected and weighted, biochemical and inflammatory profiles were determinate as well. Animals that received the pulp of Camu-camu reduced their weights of the fat in white adipose tissues, glucose, total cholesterol, triglycerides, LDL-c and insulin blood levels. There was an increase in HDL-c levels. No change was observed in inflammatory markers and liver enzymes. Camu-camu pulp was able to improve the biochemical profile of obesity in rats suggesting that this Amazonian fruit can be further used such a functional food ingredient in control of chronic diseases linked to obesity.

Technological development of aqueous extracts from Calycophyllum spruceanum (Benth.) Hook. f. ex K. Schum., Rubiaceae, (mulateiro) using factorial design

The aim of the present work is to characterize the vegetal raw material of the Calycophyllum spruceanum (Benth.) Hook. f. ex K. Schum., Rubiaceae, known as "mulateiro", and to evaluate the influence of extractive parameters for attainment of standardized aqueous extractive solutions. The physical-chemical characterization of the samples was performed using pharmacopoeic and not pharmacopoeic methodologies. A 2³ factorial design was used to evaluate the influence of extraction techniques (infusion/decoction), drug: solvent ratio (2.5 and 7.5 percent), and extraction time (5 and 15 min) on the total tannin content of aqueous extractive solutions from C. spruceanum. The extractive solution that showed higher total tannin and dry residue content had their physical-chemical characteristics determined. The results suggest that an aqueous extractive solution from rinds of C. spruceanum barks with higher tanning yield (9.9 g percent), must be standardized using decoction as extraction methodology, with 2.5 percent of vegetal drug for an extraction time of 15 min. The results of the physical-chemical characterization suggest that environmental factors modify the properties of this species and, therefore, they can influence the quality and security of a product derived from this medicinal plant.