Eduardo de Jesus Oliveira

Ocimum gratissimum Linn. and rosmarinic acid, attenuate eosinophilic airway inflammation in an experimental model of respiratory allergy to Blomia tropicalis

Allergic asthma has emerged as an important public health problem of urban populations in developed countries. Very often herbal medicine is used to treat this widespread disease, due to the lack of efficacy and the important side effects related to the classical drugs in use. Along this line, Ocimum gratissimum (Og) is a plant widely used in Brazilian folk medicine to treat inflammatory disorders, such as asthma. In the present study we evaluated the immunomodulatory effects of Og and rosmarinic acid (RA, a polyphenolic compound) in a murine model of respiratory allergy induced by the Blomia tropicalis (Bt) mite. The respiratory allergy was induced in A/J mice by administration of Bt extract and the treatment was done using 25, 50 or 100mg/kg of an Og methanolic extract or using 2, 20 or 200mg/kg of RA. We then evaluated the changes induced by these drugs on immunological parameters related to the allergic process, which are up-regulated in this allergic model. The treatment of animals with 100mg/Kg Og and 200mg/Kg RA led to a significant reduction in the numbers of leukocytes/eosinophils in bronchoalveolar lavage (BAL); eosinophil peroxidase activity in BAL; presence of mucus in respiratory tract, histopathological changes in the lung, and IL-4 in BAL. These results suggest that the methanolic extract of Og and the polyphenol RA have therapeutic potential in this murine model of respiratory allergy to a clinically relevant human sensitizer allergen.

Isoflavone formononetin from red propolis acts as a fungicide against Candida sp.

A bioassay-guided fractionation of two samples of Brazilian red propolis (from Igarassu, PE, Brazil, hereinafter propolis 1 and 2) was conducted in order to determine the components responsible for its antimicrobial activity, especially against Candida spp. Samples of both the crude powdered resin and the crude ethanolic extract of propolis from both locations inhibited the growth of all 12 tested Candida strains, with a minimum inhibitory concentration of 256 μg/mL. The hexane, acetate and methanol fractions of propolis 1 also inhibited all strains with minimum inhibitory concentration values ranging from 128 to 512 μg/mL for the six bacteria tested and from 32 to 1024 μg/mL for the yeasts. Similarly, hexane and acetate fractions of propolis sample 2 inhibited all microorganisms tested, with minimum inhibitory concentration values of 512 μg/mL for bacteria and 32 μg/mL for yeasts. The extracts were analyzed by HPLC and their phenolic profile allowed us to identify and quantitate one phenolic acid and seven flavonoids in the crude ethanolic extract. Formononetin and pinocembrin were the major constituents amongst the identified compounds. Formononetin was detected in all extracts and fractions tested, except for the methanolic fraction of sample 2. The isolated isoflavone formononetin inhibited the growth of all the microorganisms tested, with a minimum inhibitory concentration of 200 μg/mL for the six bacteria strains tested and 25 μg/mL for the six yeasts. Formononetin also exhibited fungicidal activity against five of the six yeasts tested. Taken together our results demonstrate that the isoflavone formononetin is implicated in the reported antimicrobial activity of red propolis.

Short-term stability studies of ampicillin and cephalexin in aqueous solution and human plasma: Application of least squares method in Arrhenius equation

A limited number of studies with application of the Arrhenius equation have been reported to drugs and biopharmaceuticals in biological fluids at frozen temperatures. This paper describes stability studies of ampicillin and cephalexin in aqueous solution and human plasma applying the Arrhenius law for determination of adequate temperature and time of storage of these drugs using appropriate statistical analysis. Stability studies of the beta-lactams in human plasma were conducted at temperatures of 20°C, 2°C, -20°C and also during four cycles of freeze-thawing. Chromatographic separation was achieved using a Shimpak C(18) column, acetonitrile as organic modifier and detection at 215nm. LC-UV-MS/MS was used to demonstrate the conversion of ampicillin into two diastereomeric forms of ampicilloic acid. Stability studies demonstrated degradation greater than 10% for ampicillin in human plasma at 20°C, 2°C and -20°C after 15h, 2.7days, 11days and for cephalexin at the same temperatures after 14h, 3.4days and 19days, respectively, and after the fourth cycle of freezing-thawing. The Arrhenius plot showed good prediction for the ideal temperature and time of storage for ampicillin (52days) and cephalexin (151days) at a temperature of -40°C, but statistical analysis (least squares method) must be applied to avoid incorrect extrapolations and estimated values out uncertainty limits.

A Validated Method for the Simultaneous Quantitation of Bioactive Alkaloid Markers in the Leaf Ethanolic Extract of Cissampelos sympodialis Eichl.: a Phenological Variation Study

INTRODUCTION The leaf hydroalcoholic extract of Cissampelos sympodialis Eichl. (Menispermaceae) has shown promising activity in different animal models of asthma. Several alkaloids have been identified in the extract, including warifteine and methylwarifteine (bisbenzylisoquinoline), as well as milonine (morphinandienone). OBJECTIVE To develop and validate an analytical method for the simultaneous quantitation of the bioactive markers of C. sympodialis hydroalcoholic leaf extract and to apply the method to a seasonal (phenological) study of the concentration of the alkaloid markers. METHODOLOGY The method used reversed phase high performance liquid chromatography with UV detection and calibration by standard addition. Separation was achieved using a C₁₈-column (250 × 4.6 mm, 5 µm) and a mobile phase consisting of a mixture of 0.05% aqueous (Et)₃NH₂ (A):MeOH(B) in gradient mode at a flow rate of 1.0 mL/min. RESULTS The method proved to be linear in the concentration range tested (2-100 µg/mL, r² > 0,99), precise (RSD ≤ 15%), accurate (85-115%), selective and robust. Detection limits for warifteine, methyl-warifteine and milonine were 0.39, 1.10 and 1.77 µg/mL respectively. The highest concentration of total alkaloids (determined as the sum of the three alkaloids) in the hydroalcoholic extract of the leaves was 2.9 ± 0.2 mg/g extract (n = 3), prior to fruit development. Both warifteine and methylwarifteine were detected in the total alkaloid fraction of the ripened fruits. CONCLUSION The results demonstrated that significant variations in the concentration of the biomarkers occurred throughout the vegetative cycle. The lowest concentration of the alkaloids in the leaves coincided with their appearance in the ripened fruits.

Usefulness of pyrolysis coupled to gas chromatography/mass spectrometry for evaluating the reproducibility of commercial samples of Cymbopogon citratus Stapf., Poaceae

The usefulness of pyrolysis coupled to gas chromatography/mass spectrometry (Py-GC/MS) to evaluate the identity and reproducibility of different brands and batches of commercially available samples of Cymbopogon citratus Stapf (sold as tea) was investigated. Samples of the vegetable material were extracted using hexane and the extract was pyrolysed at 450 oC using a vertical microfurnace pyrolyser interfaced directly with a gas chromatograph/mass spectrometer operated using electron impact ionization at 70 eV. The relative peak areas of the different compounds identified on the pyrograms were used for multivariate data analysis using principal component analysis (PCA) and hierarchical cluster analysis (HCA) techniques. Principal component analysis of pyrolysis data from samples of different brands was able to represent 84.7% of the total variability within the first two principal components, and led to the correct classification of the samples tested. The method was also able to reveal important differences between samples when different batches of the same brand were compared.

Distinct effects of novel naphtoquinone-based triazoles in human leukaemic cell lines

The aim of this study was to investigate the cytotoxic effect of new 1,4‐naphthoquinone‐ 1,2,3‐triazoles, named C2 to C8 triazole derivatives, towards human cancer cell lines.

Complete ¹H and 13C NMR assignments of isojuripidine from Solanum asterophorum Mart.

Isojuripidina, um alcaloide 3-aminoespirosolano foi isolado das partes aereas de Solanum asterophorum Mart. A estrutura foi determinada usando uma combinacao de tecnicas de RMN homo (1D: RMN 1H, RMN 13C{1H}, RMN13C-DEPT135; 2D: COSY, 1H-1H-NOESY) e heteronuclear 2D (HSQC e HMBC) e espectro de massas de alta resolucao. Foi feita tambem a atribuicao inequivoca dos deslocamentos quimicos dos atomos de carbono e hidrogenio dos derivados 3-N,6-O-diacetil-isojuripidina e 3-N-cinamoil-isojuripidina.

Studies with Cissampelos sympodialis: the search towards the scientific validation of a traditional Brazilian medicine used for the treatment of asthma

The paper is a review of the literature on the ethnobotanical, chemical and pharmacological aspects of the species Cissampelos sympodialis Eichler, Menispermaceae, in order to assess its potential for the treatment of asthma. The aqueous infusion from the leaves of this Brazilian plant is used in the traditional medicine for the treatment of respiratory conditions, including colds, bronchitis and asthma. A multidisciplinary approach has led to the elucidation of the main chemical biomarkers and of the mechanism of action of the extract and its isolated constituents in animal models of inflammation and asthma. A comprehensive review of the literature on the species and its related chemical constituents was conducted using Pubmed, Web of Sciences, Lilacs, SciFinder, as well as conference proceedings. Retrieved literature data demonstrates that the aqueous fraction of the ethanolic extract from the leaves exerts an immunomodulatory activity in different animal models of asthma. This include an increase in the levels of anti-inflammatory cytokines, a decrease in the production of antigen-specific immunoglobulin, a decrease in mucus production and deposition in the airways, and a direct bronchodilator activity. These preclinical results clearly demonstrate the potential of this species for the treatment of asthma and points to the need for well-designed clinical trials to finally validate the traditional use of this herbal medicine.

Antioxidant and Antihypertensive Effects of a Chemically Defined Fraction of Syrah Red Wine on Spontaneously Hypertensive Rats

A particularly phenolic-rich fraction extracted from red wine from the São Francisco valley (Northeastern Brazil) was chemically characterized and its hypotensive and antioxidant effects on spontaneously hypertensive rats were studied both in vitro and in vivo. The liquid-liquid pH dependent fractionation scheme afforded a fraction with high content of bioactive phenolics such as flavonols, flavonol glycosides, phenolic acids and anthocyanins, whose identities were confirmed by liquid chromatography coupled to mass spectrometry analysis. Pretreatment of spontaneously hypertensive rats with this wine fraction at doses of 50 and 100 mg/kg by gavage for 15 days was able to decrease mean arterial pressure and heart rate as well as decrease serum lipid peroxidation. The fraction at concentrations of 0.01–1000 µg/mL induced concentration-dependent relaxation of isolated rat superior mesenteric artery rings pre-contracted with phenylephrine and this effect was not attenuated by endothelium removal. Our results demonstrate it is possible for phenolic constituents of red wine that are orally bioavailable to exert in vivo hypotensive and antioxidant effects on intact endothelial function.

1H and 13C NMR assignments of two new isomeric bisbenzylisoquinoline alkaloids from Cissampelos sympodialis Eichl. (Menispermaceae).

des‐7′‐O‐methylroraimine (compound 1) and epi‐des‐7′‐O‐methylroraimine (compound 2), two new isomeric bisbenzylisoquinoline alkaloids were isolated and characterized as a mixture from the rhizomes of Cissampelos sympodialis Eichl. The unambiguous structural elucidation of both isomers was performed with the aid of HR‐ESI‐MS, FT‐IR, and NMR techniques including COSY, HMQC, HMBC, and NOESY. Copyright