Naringenin is a flavonoid with several biological activities already reported but with few biological applications in the pharmaceutical field. In this work, a new flavanone, called carboxymidamide (DCHA), synthesized through the condensation reaction between naringenin and aminoguanidine is structurally confirmed through spectroscopic analysis of nuclear magnetic resonance, mass spectrometry and Fourier transform infrared. DCHA was not toxic to human fibroblasts and inhibited macrophage activation in vitro. In vivo, it suppressed leukocyte migration in lipopolysaccharide (LPS)-induced peritonitis and reduced zymosan-induced paw edema. Molecular docking simulations of DCHA in the active site of the enzymes inducible nitric oxide synthase, cyclooxygenase-2 and phosphodiesterase type 4 indicate that the anti-inflammatory activity of this new flavanone can be explained by the inhibition of these target enzymes. This is the first work to present the synthesis of a flavonoid derivative with aminoguanidine that showed a high anti-inflammatory potential. field DCHA, , (DCHA) resonance infrared vitro vivo LPSinduced LPS induced zymosaninduced zymosan edema synthase cyclooxygenase2 cyclooxygenase 2 cyclooxygenase- antiinflammatory anti inflammatory potential (DCHA

A novel anti-inflammatory hybrid 3-ibuprofenyl-copalic acid (3-IbuCA) was synthesized from 3-hydroxy-copalic acid isolated from Amazonian copaiba oil (Copaifera multijuga Hayne), and the anti-inflammatory ibuprofen. After full characterization, several assays to verify its anti-inflammatory effects were performed in vitro, in vivo and in silico (molecular docking). Induced fit docking was performed to observe the interactions with the enzymes cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2). In vitro tests of cytotoxicity and tumor necrosis factor (TNF)-α inhibition, and in vivo tests of pleurisy, protein expression and gastrocytotoxicity were performed. Molecular docking studies with COX-1 and 2 showed binding free energies (ΔG) of -2.2 and -7.8 kcal mol-1, respectively, while for mofezolac and indomethacin, the binding free energies ΔG presented values of -8.5 and -10.1 kcal mol-1, which makes 3-IbuCA selective for COX-2 inhibition. This hybrid showed no toxicity against human macrophage at concentrations up to 2 µM, and inhibited TNF-α production in lipopolysaccharide (LPS)-stimulated macrophages. In the pleurisy assays, 3-IbuCA reduced the total leukocytes and mononuclear cells, which was followed by reduction of p-IKBα (phosphorylated nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor, alpha) protein expression. Compared with ibuprofen alone, the hybrid caused less gastric damage. Thus, the docking, together with in vitro and in vivo studies suggest that this novel hybrid has potential as a new anti-inflammatory agent.

Phytochemical investigation of Endopleura uchi led to the isolation of a gallic acid derivate, known as bergenin (2) and friedelin (1), a pentacyclic triterpene. The present work also reports the bergenin quantification in different Endopleura uchi bark, twig and leaves extracts. Our findings showed the highest bergenin concentration in bark methanol extracts (4.75%) and the lowest concentration in twig aqueous extracts (1.89%). Phenolics quantification by Folin-Ciocalteu, revealed phenolic compounds level values from 16.69 to 43.02 mg GAE g-1 dry extract. The ferric reducing antioxidant activity ranged from 230.43 to 567.89 mmol Fe+2 g-1 dry extract. DPPH IC50 free radicals showed to range from 12.04 to 24.20 μg mL-1. Leaves’ chloroform fraction exhibited the highest nitric oxide inhibiting activity bearing IC50= 3.2 μg mL-1. Results show this species to hold significant bergenin concentrations as well as phenolic and anti-inflammatory compounds in all extracts suggesting it bear therapeutic potential. derivate 2 (2 1, 1 , (1) triterpene 4.75% 475 4 75 (4.75% 1.89%. 189 1.89% . 89 (1.89%) FolinCiocalteu, FolinCiocalteu Folin Ciocalteu, Ciocalteu Folin-Ciocalteu 1669 16 69 16.6 4302 43 02 43.0 g1 g g- extract 23043 230 230.4 56789 567 567.8 Fe2 Fe Fe+ IC IC5 1204 12 04 12.0 2420 24 20 24.2 mL1. mL1 mL 1. mL-1 Leaves 32 3 3. antiinflammatory anti inflammatory potential ( (1 4.75 47 7 (4.75 18 1.89 8 (1.89% 166 6 16. 430 0 43. 2304 23 230. 5678 56 567. 120 12. 242 24. mL- 4.7 (4.7 1.8 (1.89 5 4. (4. (1.8 (4 (1.

Piquiá (Caryocar villosum - Caryocaraceae) is a native fruit from the Amazon region rich in bioactive substances. Fruit pulp extracts were analyzed by HPLC, together with extracts obtained from fruit pulp industry residual parts, byproducts such as husks (shells) and seeds. Extracts were prepared with two ethanolic solvent systems. Phenolic substances ellagic and gallic acids were detected with standards and quantified by HPLC. Cytotoxic, antioxidant and anti-inflammatory activities in vitro were also evaluated. Shell extracts showed free radicals scavenger capacity in ABTS (IC50: 3.93 ± 0.12 µg mL-1) and DPPH models (IC50: 7.81 ± 0.34 µg mL-1), low cytotoxicity in human fibroblasts, but high at tumor strains, and also a high anti-inflammatory potential observed by the inhibition of nitric oxide production. At low concentrations (20 µg mL-1), excellent antioxidant activities were verified in cellular assays, with percentages of 70.69 ± 2.77%, 79.89 ± 6.50% and 79.48 ± 8.6% for shell, pulp and seed extracts, respectively. With this set of results, C. villosum fruit extracts become a high potential raw material to be used in pharmaceutical and cosmetic applications.

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.

O fruto Camu-camu da amazônia (Myrciaria dubia HBK Mc Vaugh) apresenta um interesse atrativo na alimentação e industria de cosméticos devido ao rico conteúdo de Vitamina C, flavonoides e antocianinas. O objetivo deste estudo foi investigar a ação antiobesidade da ingestão da polpa de Camu-camu em modelo de ratos obesos. Ratos Wistars foram induzidos a obesidade pela injeção subcutânea de glutamato monossódico recebendo dieta ad libitum. Os ratos foram divididos em dois grupos. Um grupo experimental, que ingeriram 25 mL/dia de polpa de Camu-camu (CCG) e um grupo não tratado (CG). Após 12 semanas, os animais foram sacrificados. Sangue, fígado, coração e tecido adiposo foram coletados e pesados, assim como determinado os perfis bioquímicos e inflamatórios. Animais que receberam a polpa de Camu-camu reduziram o peso do tecido adiposo, os níveis plasmáticos de glicose, colesterol total, trigliceridios, LDL-c e níveis de insulina no sangue. Houve um aumento nos níveis de HDL-c. Nenhuma mudança foi observada nos marcadores inflamatórios e enzimas do fígado. A polpa de Camu-camu foi capaz de melhorar o perfil bioquímico da obesidade em ratos, sugerindo que este fruto da Amazônia pode ser futuramente utilizado como um ingrediente alimentar funcional no controle de doenças crônicas ligadas a obesidade.

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.

O lúpus eritematoso sistêmico (LES) é uma doença autoimune de origem desconhecida, associada ao estresse oxidativo. O presente estudo teve como objetivo investigar a presença de estresse oxidativo em pacientes com LES recém-diagnosticado. Pacientes com LES (n = 36) e controles (n = 28) foram incluídos no estudo. Amostras de sangue foram usadas para dosagem de malondialdeído (MDA), grupo sulfidrila (SH) e ácido úrico no soro. Os níveis de MDA (µmol/L) foram maiores nos pacientes (3,9 ± 2,6) que nos controles (1,6 ± 2,6). Os níveis de SH foram significativamente menores nos pacientes. Os achados sugerem que o MDA pode ser um bom marcador de estresse oxidativo no LES.

Systemic lupus erythematosus (SLE) is an autoimmune disease of unknown origin associated with oxidative stress. The present study aimed to investigate the presence of oxidative stress in patients with newly diagnosed SLE. SLE patients (n = 36) and control subjects (n = 28) were enrolled in this study. Blood samples were used for malondialdehyde (MDA), sulfhydryl groups (SH) and uric acid determination. MDA levels (µmol/L) were higher in patients (3.9 ± 2.6) than in control subjects (1.6 ± 2.6). SH were significantly lower in SLE patients. The findings suggest that MDA can be a good marker of oxidative stress in SLE.

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%), 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%), must be standardized using decoction as extraction methodology, with 2.5% 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.

Malaria remains an important health problem in tropical countries like Brazil. Thrombocytopenia is the most common hematological disturbance seen in malarial infection. Oxidative stress (OS) has been implicated as a possible mediator of thrombocytopenia in patients with malaria. This study aimed to investigate the role of OS in the thrombocytopenia of Plasmodium vivax malaria through the measurement of oxidant and antioxidant biochemical markers in plasma and in isolated platelets. Eighty-six patients with P. vivax malaria were enrolled. Blood samples were analyzed for total antioxidant and oxidant status, albumin, total protein, uric acid, zinc, magnesium, bilirubin, total thiols, glutathione peroxidase (GPx), malondialdehyde (MDA), antibodies against mildly oxidized low-density lipoproteins (LDL-/nLDL ratio) and nitrite/nitrate levels in blood plasma and GPx and MDA in isolated platelets. Plasma MDA levels were higher in thrombocytopenic (TCP) (median 3.47; range 1.55-12.90 µmol/L) compared with the non-thrombocytopenic (NTCP) patients (median 2.57; range 1.95-8.60 µmol/L). Moreover, the LDL-/nLDL autoantibody ratio was lower in TCP (median 3.0; range 1.5-14.8) than in NTCP patients (median 4.0; range 1.9-35.5). Finally, GPx and MDA were higher in the platelets of TPC patients. These results suggest that oxidative damage of platelets might be important in the pathogenesis of thrombocytopenia found in P. vivax malaria as indicated by alterations of GPx and MDA.

A hanseníase, doença infecciosa crônica, é causada pelo Mycobacterium leprae. Depois da Índia, o Brasil possui o segundo maior número de casos no mundo. O aumento do estresse oxidativo e da deficiência das defesas antioxidantes estão presentes em indivíduos infectados e podem associar-se à progressão da infecção. Foram estudadas alterações nos níveis séricos da peroxidação lipídica e vitamina A em pacientes com diferentes formas de hanseníase. Foram selecionados para o estudo quatro grupos de pacientes com hanseníase e um grupo controle (indivíduos saudáveis) e os níveis séricos de vitamina A e a peroxidação lipídica, medida através do malondialdeído (MDA), foram determinados por métodos espectrofotométricos. Os níveis séricos médios de MDA (µmol/L) foram 3,80 ± 0,5 no grupo controle e 10,54 ± 1,1 nos pacientes com hanseníase. Sendo este aumento gradual e exacerbado nas formas mais severas da doença. Quanto à vitamina A, os níveis séricos (µg/dL) encontraram-se diminuídos nos indivíduos infectados (38.51 ± 4.2), principalmente na forma lepromatosa, quando comparados com o grupo controle (53.8 ± 5.6). Estes resultados indicam que a peroxidação lipídica pode ser um fator importante na infecção mediada pelo Mycobacterium leprae podendo estar relacionada ao aumento da atividade fagocítica pelos macrófagos contribuindo para um aumento da LPO durante a progressão da infecção. A avaliação do perfil oxidante/antioxidante nestes pacientes pode ser um fator importante no tratamento, controle e/ou prognóstico desta doença.

Leprosy, a chronic infectious disease, is caused by a Mycobacterium leprae infection. After India, Brazil has the second greatest number of cases in the world. Increase of oxidative stress and antioxidant deficiency are present in infected subjects and can be related to infection progression. We studied alterations in serum levels of lipid peroxidation (LPO) and vitamin A in patients with different forms of leprosy. Four groups of leprosy patients and a control group (healthy subjects) were selected, and their vitamin A serum levels and LPO profile, measured as malonaldehyde (MDA) were measured by spectrophotometric assays. The mean MDA serum levels (µmol/L) were 3.80 ± 0.5 for control group and 10.54 ± 1.1 in the leprosy patients and this increase was gradual, being more accentuated in severe forms of the disease. Also, the vitamin A serum levels (µg/dL) were diminished in the infected subjects (38.51 ± 4.2), mainly in lepromatous form, when compared with the control group (53.8 ± 5.6). These results indicate that LPO can be an important factor in Mycobacterium leprae infection, which can be related to increases in phagocytic activity and the general breakdown of antioxidants, contributing to an increase of LPO during infection progression. The evaluation of oxidant/antioxidant status in these patients can be an important factor in the treatment, control, and/or prognosis of this disease.

Os tióis são descritos como os principais responsáveis pelos efeitos antioxidantes das proteínas plasmáticas. Além disso, diversos estudos mostram uma correlação positiva entre os níveis séricos de tióis e a capacidade antioxidante total (CAT). A medida da CAT por métodos baseados na oxidação de substratos tem sido amplamente usada como referência na estimativa da capacidade antioxidante em amostras biológicas; porém, em muitos casos esses métodos são inexatos e imprecisos, principalmente quando realizados por procedimentos não-automatizados. Neste artigo descrevemos um procedimento automatizado simples para a determinação de tióis totais no soro, com base no conhecido método de Ellman. A dosagem dos tióis foi correlacionada com diversos marcadores da capacidade oxidante/antioxidante, como a CAT, o teste das substâncias reativas ao ácido tiobarbitúrico (TBARs) e os níveis de peróxidos totais. Os tióis correlacionaram-se positivamente com a CAT (r = 0,298; p < 0,001) e negativamente com os níveis de TBARs (r = - 0,330; p < 0,001). O procedimento aqui descrito para a dosagem de tióis pode ser uma ferramenta importante na medida da capacidade antioxidante juntamente com a dosagem da CAT. A utilização desse novo procedimento possibilita a determinação dos tióis totais em larga escala por laboratórios clínicos e de pesquisa onde essa demanda é necessária.

Thiol groups have been described as the main responsible for antioxidative effects of plasmatic proteins. Also, thiol serum levels have shown a positive correlation with total antioxidant capacity (TAC) in many studies. Measurement of TAC by substract oxidation-based methods have been widely used as a reference to measure antioxidant status; however, in many cases these methods are inexact or imprecise, usually when performed by manual procedures. In this paper we describe a simple automated procedure for the determination of total thiols in serum, which was based on Ellman’s method. It was correlated with several markers of oxidative/antioxidative status, such as TAC and thiobarbituric acid reactive substance test (TBARs). Serum thiol levels were correlated positively with TAC (r = 0.298, p < 0.001) and negatively with TBARs levels (r = -0.330, p < 0.001). The novel automated procedure for thiol groups measurement can be a great tool in estimation of antioxidant status together with TAC assay. This procedure makes the determination of total thiol groups in large scale possible in clinical chemistry or research laboratories where this approach is necessary.