Biological effects of extracts obtained from Stryphnodendron adstringens on Herpetomonas samuelpessoai

We report the effect of Stryphnodendron adstringens on the trypanosomatid Herpetomonas samuelpessoai. The parasites were grown at 28 degrees C in a chemically defined medium containing crude extract and fractions at concentrations from 100 to 5000 microg/ml obtained from S. adstringens. Concentrations of 500, 1000, 2500, and 5000 microg/ml both crude extract and semi-purified fraction progressively inhibited the protozoans' growth. At a concentration of 100 microg/ml, crude extract or a semi-purified (F3) fraction did not affect the growth of the protozoans. The F3-9 - F3-12 sub-fractions, at a concentration of 1000 microg/ml, also showed increased inhibitory activity on H. samuelpessoai. The IC50 of the crude extract and the F3 fraction were 538 and 634 microg/ml, respectively. Ultrastructural and enzymatic alterations in the trypanosomatids were also evaluated. H. samuelpessoai cultivated in the presence of IC50 crude extract showed considerable ultrastructural alterations, such as marked mitochondrial swelling with a large number of cristae and evident Golgi complex vesiculation, as observed by transmission electron microscopy. Cells exposed to 538 microg/ml of crude extract at 28 degrees C for 72 h, showed decreased activity of the enzyme succinate cytochrome c reductase, a typical mitochondrion marker, as compared to untreated cells.     

Anti-herpesvirus bovine type 5 activities of extracts obtained from Plocamium brasiliense

Bovine herpesvirus type 5 (BoHV-5) is an important etiologic agent of meningoencephalitis in cattle and has been frequently identified in outbreaks of neurological disease in bovine in the southern hemisphere including Brazil. This study aimed to evaluate the cytotoxic effect and the antiviral properties of extracts obtained from Plocamium brasiliense (Greville) Howe and Taylor in BoHV-5 RJ42/01 replication. The cytotoxic effects were measured in Madin-Darbin bovine kidney cells (MDBK) and cytotoxic concentration (CC₅₀) values have been determined for acyclovir (ACV) (223 μg?±?2.0), ethyl acetate extract from P. brasiliense (2,109 μg?±?10), hexane extract from P. brasiliense (7.181 μg?±?5), dichloromethane extract from P. brasiliense (2.356 μg?±?6.5), and hydroalcoholic extract from P. brasiliense (1.408 μg?±?5.8). As a first approach to characterize the action of these extracts on BoHV-5 RJ42/01, a virucidal assay activity was performed. A virus suspension containing 1?×?10⁵ plaque-forming units (PFU) of the BoHV-5 RJ42/01 was mixed with 600 μg of extract and acyclovir and kept at room temperature (24 °C) for 3 h. Meanwhile, a control of untreated infected virus was performed in the same conditions. Then, treated virus suspension and untreated control were diluted, and percentage of inhibition of infectivity was determined by plaque assay: ethyl acetate extract (99 %), hexane extract (90 %), dichloromethane extract (99 %), and hydroalcoholic extract (27 %). Acyclovir had a slight virucidal activity on viral particle. The inhibition of attachment was performed in MDBK cells inoculated with 100 PFU of BoHV-5 RJ42/01 in the presence or absence of various concentrations of extracts (0.3, 0.9, and 1.5 μg mL^?1). Acyclovir was also assayed at 2.8 and 11.25 μg mL^?1. The inhibition of adsorption was also tested in MDBK cells treated with the same concentrations of the extracts before virus inoculation. Results: hexane extracts inhibited virus attachment in pre-treated cell 0.9 μg (55 %) and 1.5 μg (71 %) and untreated MDBK cell only with 1.5 μg (63 %). Ethyl acetate extract on cell pre-treated with 0.3 μg (67 %), 0.9 μg (81 %), and 1.5 μg (91 %). Ethyl acetate extract on pre-treated cell 0.3 μg (67 %), 0.9 μg (81 %), and 1.5 (91 %) but discrete inhibition on cell untreated. Dichloromethane extract and acyclovir slightly inhibited virus binding on MDBK cell.     

Anti-leishmanial activity of alkaloidal extracts obtained from different organs of Aspidosperma ramiflorum

The present study was designated to evaluate semi-quantitative antileishmanial activity of alkaloidal extracts that were obtained from 1g of different parts of Aspidosperma ramiflorum (leaves, roots, seeds, and stem barks). Alkaloidal extracts of barks and leaves presented a good activity against the extracellular form (promastigotes) of Leishmania (L.) amazonensis. It is known that compounds responsible for the antileishmanial activity in the alkaloidal extracts from A. ramiflorum are the monoterpenoid indole alkaloids ramiflorine A and ramiflorine B, therefore extracts obtained from different plant parts were analyzed by electrospray ionization mass spectrometry (ESI-MS) in order to evidence the presence of these bioactive alkaloids. Based on these findings, alkaloidal extract from leaves was fractionated on preparative thin-layer chromatography in a bioassay-guided fractionation affording individual purified ramiflorines A and B. Both ramiflorines A and B showed significant activity against Leishmania (L.) amazonensis (LD(50) values of 18.5±6.5μg/ml and 12.63±5.52μg/ml, respectively). Our results are showing that alkaloidal extract from leaves is a promising alternative to the use of stem barks from A. ramiflorum.     

Cytohistological and phytochemical study of madder root extracts obtained by ultrasonic and classical extractions

Introduction – Madder (Rubia tinctorum) has been used since ancient times as a source of pigments for dyeing and painting. Madder dyes are localised in roots and the native chemical population is composed of glycosiled and aglycone compounds. The aim of this study is to elaborate an efficient extraction process without any chemical denaturation of dyes. Objective – To compare an optimised ultrasonic process, using for madder dye extraction, with two conventional procedures and to determine the efficiency of ultrasound on these vegetable matrix. Methodology – Madder roots were extract in a methanol–water mixture in 37 : 63 (v/v) for ultrasound and 80 : 20 (v/v) for reflux and agitation. HPLC‐PAD analyses showed the anthraquinone proportion for each extraction process and their denaturing effects. Finally, cytohistological observations were made to show the consequence of each process on the cell organisation in madder roots. Results – The results showed that the amount of extracted dyes was higher with UAE than with agitation and reflux. HPLC‐PAD analysis revealed that the anthraquinone composition differed according to the extraction procedure. The UAE extracts presented an important richness in terms of anthraquinonic compounds that suggests a preserving effect. Cytohistological observations showed that the main alterations concerned the cell walls of phloem. After UAE the walls exhibited numerous pitted areas reflecting an ultrasound‐induced cavitation that enhances the extraction effectiveness of this method. Conclusion – The study has shown the improvement of madder roots extraction both quantitatively and qualitatively using the efficiency of ultrasound‐assisted extraction in comparison with magnetic agitation and reflux techniques. Copyright © 2009 John Wiley & Sons, Ltd.     

Artemisinin content in Artemisia annua L. extracts obtained by different methods

Isolation of artemisinin from Artemisia annua L. and its quantification by the HPLC-MS method are considered. Different extraction methods were used for the isolation of artemisinin: maceration, ultrasonic and subcritical CO₂-extraction. The component content of the CO₂- and hexane extracts was studied by the GC-MS method.     

Determination of phenolic content and antioxidant activity of extracts obtained from Rosmarinus officinalis' calli

Rosmarinus officinalis is widely found in the lands of Aegean and Mediterranean regions of Turkey. Stem explants of very young shoots were cultured in both woody plant medium (WPM) and Murashige and Skoog (MS) media supplemented with 7g/L agar, 30g/L sucrose, and 1 and 3mg/L naphthaleneacetic acid (NAA) for callus initiation. Induced calli were subcultured 4 times with intervals of 7-10 days. MS medium supplemented with 1mg/L NAA proved to be the best medium for the production of callus (65.0%) among the samples tested. The lyophilized calli were subjected to solvent extraction. Active constituents of 8 calli extracts were analyzed by HPLC, and rosmarinic acid (RA) was determined to be the primary compound. Calli cultivated in WPM supplemented with 1mg/L NAA and extracted at 50 degrees C, yielded the highest amount of RA (34.4mg/g dry weight). Moreover, antioxidant activity of calli extracts was determined using a number of in vitro assays, including total phenol assay, DPPH radical scavenging activity (RSA), and trolox equivalent antioxidant capacity (TEAC). On the basis of the current findings, we conclude that WPM supplemented with 1mg/L NAA yields higher phenolic content as well as higher antioxidant activity.     

Immunologic study of artificial complex antigens obtained from P. aeruginosa lipopolysaccharides]

Artificial antigens were obtained on the basis of the polysaccharide component of P. aeruginosa complexed with an indifferent protein. Immunological study indicated that the specific polysaccharide of P. aeruginosa lipopolysaccharide contained two structures, high molecular and low molecular, having qualitative and quantitative differences in their hydrocarbon composition. Artificial complex antigens possessed serological and immunogenic properties, the low molecular polysaccharide fraction complexed with protein having less pronounced serological and immunogenic activity than polysaccharide and the high molecular fraction complexed with protein. Antificial complex antigens exerted no protective effect in generalized P. aeruginosa infection in rats.     

Effect of extracts obtained from olive oil mill waste waters on Bacillus megaterium ATCC 33085

n -Propanol was the most effective solvent for extracting antibacterial substances from olive oil mill waste water ('alpechines'). Several phenolics were detected in propanol extracts that had bactericidal effects on Bacillus megaterium ATCC 33085, inhibiting sporulation and germination at 5.6 mmol/l total phenolics (expressed as syringic acid). The biological effect was increased in the presence of high glucose and NaCl concentrations and after β-glucosidase hydrolysis.     

Preliminary investigation on chemical composition and bioactivity of differently obtained extracts from Symphytum aintabicum Hub.- Mor. &Wickens

Symphytum species, commonly known as comfrey, hold a long history of folk medicinal uses for healing wounds, bone fractures and other pain resulting from inflammation. However, many species belonging to this genus have not yet been scientifically investigated and hence investigation in this regard is warranted. In the present research, S. aintabicum extracts prepared by different extraction techniques (homogenizer assisted extraction, infusion and maceration) were assayed for their chemosystematic attributes (total phenolic, flavonoid) as well as bioactive compound contents, antioxidant and enzyme inhibition potentials. For instance, the extracts were found to possess phenolic and flavonoid levels ranging from 35.50 to 112.25 mg GAE/g and 2.54–25.12 mg RE/g, respectively. In general, the extracts displayed significant antioxidant activity. However, methanolic and infusion extracts were observed as the most potent radical scavengers and reducing agents, most likely related to their phenolic content. The extracts also showed metal chelating power and total antioxidant capacity in phosphomolybdenum assay. Moreover, while all extracts showed inhibition on amylase, only the methanolic extracts acted as inhibitors of cholinesterases and tyrosinase, most probably due to their relatively higher flavonoid content. The findings also indicated that the extraction method as well as the solvent type used to more or less affect the yield of the bioactive compounds and extracts’ bioactivity reported herein. Hence, as a first report highlighting the in vitro pharmacological properties of S. aintabicum, promising results were revealed from the quantitative chemosystematic analysis of bioactive components present and as sources of antioxidants and enzyme inhibitors against key human illnesses, thus providing scope for potential applications in phytomedicine development.