Nanoemulsions and Solid Microparticles Containing Pentyl Cinnamate to Control Aedes aegypti.

The Aedes aegypti mosquito is a vector of severe diseases with high morbidity and mortality rates. The most commonly used industrial larvicides have considerable toxicity for non-target organisms. This study aimed to develop and evaluate liquid and solid carrier systems to use pentyl cinnamate (PC), derived from natural sources, to control Ae. aegypti larvae. The liquid systems consisting of nanoemulsions with different lecithins systems were obtained and evaluated for stability over 30 days. Microparticles (MPs) were obtained by the spray drying of the nanoemulsions using maltodextrin as an adjuvant. Thermal, NMR and FTIR analysis indicated the presence of PC in microparticles. Indeed, the best nanoemulsion system was also the most stable and generated the highest MP yield. The PC larvicidal activity was increased in the PC nanoemulsion system. Therefore, it was possible to develop, characterize and obtain PC carrier systems active against Ae. aegypti larvae.

Characterization of the biochemical, physiological, and medicinal properties of Streptomyces hygroscopicus ACTMS-9H isolated from the Amazon (Brazil).

Actinomycetes are known to produce numerous secondary bioactive metabolites of pharmaceutical interest. The purpose of this study was to isolate, characterize, and investigate the antibacterial, antifungal, and anticancer activities of metabolites produced by Actinobacteria isolated from the rhizosphere of Paullinia cupana. The Actinobacteria was identified as Streptomyces hygroscopicus ACTMS-9H. Based on a bioguided study, the methanolic biomass extract obtained from submerged cultivation had the most potent antibacterial, antifungal, and cytotoxic activities. This extract was partitioned with n-hexane, ethyl acetate, and 2-butanol. Elaiophylin was isolated from the methanolic biomass extract, and its molecular formula was determined (C54H88O18) based on 1H and 13C NMR, IR and MS analyses. The 2-butanol phase was fractionated into four fractions (EB1, EB2A, EB2B, and EB3M). Chemical prospecting indicated the presence of alkaloids, saponins, and reducing sugars in the methanolic extract and 2-butanol phase. The elaiophylin displayed anticancer activity in HEp-2 and HL-60 cells with an IC50 of 1 µg/mL. The EB1 fraction was selectively toxic to HL-60 cells with IC50 of 9 ng/mL. Bioautography showed that the EB1 fraction contained an alkaloid with antibacterial and antifungal activities (MIC values ≤1.9 and <3.9 µg/mL, respectively). In conclusion, the EB1 fraction and elaiophylin of S. hygroscopicus have potent antimicrobial, antifungal, and anticancer activities.

Inhibition of Neurotoxic Secretory Phospholipases A(2) Enzymatic, Edematogenic, and Myotoxic Activities by Harpalycin 2, an Isoflavone Isolated from Harpalyce brasiliana Benth.

Secretory phospholipases A(2) (sPLA(2)) exert proinflammatory actions through lipid mediators. These enzymes have been found to be elevated in many inflammatory disorders such as rheumatoid arthritis, sepsis, and atherosclerosis. The aim of this study was to evaluate the effect of harpalycin 2 (Har2), an isoflavone isolated from Harpalyce brasiliana Benth., in the enzymatic, edematogenic, and myotoxic activities of sPLA(2) from Bothrops pirajai, Crotalus durissus terrificus, Apis mellifera, and Naja naja venoms. Har2 inhibits all sPLA(2) tested. PrTX-III (B. pirajai venom) was inhibited at about 58.7%, Cdt F15 (C. d. terrificus venom) at 78.8%, Apis (from bee venom) at 87.7%, and Naja (N. naja venom) at 88.1%. Edema induced by exogenous sPLA(2) administration performed in mice paws showed significant inhibition by Har2 at the initial step. In addition, Har2 also inhibited the myotoxic activity of these sPLA(2)s. In order to understand how Har2 interacts with these enzymes, docking calculations were made, indicating that the residues His48 and Asp49 in the active site of these enzymes interacted powerfully with Har2 through hydrogen bonds. These data pointed to a possible anti-inflammatory activity of Har2 through sPLA(2) inhibition.

Harpalycin 2 inhibits the enzymatic and platelet aggregation activities of PrTX-III, a D49 phospholipase A2 from Bothrops pirajai venom.

BACKGROUND: Harpalycin 2 (HP-2) is an isoflavone isolated from the leaves of Harpalyce brasiliana Benth., a snakeroot found in northeast region of Brazil and used in folk medicine to treat snakebite. Its leaves are said to be anti-inflammatory. Secretory phospholipases A2 are important toxins found in snake venom and are structurally related to those found in inflammatory conditions in mammals, as in arthritis and atherosclerosis, and for this reason can be valuable tools for searching new anti-phospholipase A2 drugs. METHODS: HP-2 and piratoxin-III (PrTX-III) were purified through chromatographic techniques. The effect of HP-2 in the enzymatic activity of PrTX-III was carried out using 4-nitro-3-octanoyloxy-benzoic acid as the substrate. PrTX-III induced platelet aggregation was inhibited by HP-2 when compared to aristolochic acid and p-bromophenacyl bromide (p-BPB). In an attempt to elucidate how HP-2 interacts with PrTX-III, mass spectrometry, circular dichroism and intrinsic fluorescence analysis were performed. Docking scores of the ligands (HP-2, aristolochic acid and p-BPB) using PrTX-III as target were also calculated. RESULTS: HP-2 inhibited the enzymatic activity of PrTX-III (IC50 11.34 ± 0.28 µg/mL) although it did not form a stable chemical complex in the active site, since mass spectrometry measurements showed no difference between native (13,837.34 Da) and HP-2 treated PrTX-III (13,856.12 Da). A structural analysis of PrTX-III after treatment with HP-2 showed a decrease in dimerization and a slight protein unfolding. In the platelet aggregation assay, HP-2 previously incubated with PrTX-III inhibited the aggregation when compared with untreated protein. PrTX-III chemical treated with aristolochic acid and p-BPB, two standard PLA2 inhibitors, showed low inhibitory effects when compared with the HP-2 treatment. Docking scores corroborated these results, showing higher affinity of HP-2 for the PrTX-III target (PDB code: 1GMZ) than aristolochic acid and p-BPB. HP-2 previous incubated with the platelets inhibits the aggregation induced by untreated PrTX-III as well as arachidonic acid. CONCLUSION: HP-2 changes the structure of PrTX-III, inhibiting the enzymatic activity of this enzyme. In addition, PrTX-III platelet aggregant activity was inhibited by treatment with HP-2, p-BPB and aristolochic acid, and these results were corroborated by docking scores.

Antimicrobial Peptide Analogs From Scorpions: Modifications and Structure-Activity.

The rapid development of multidrug-resistant pathogens against conventional antibiotics is a global public health problem. The irrational use of antibiotics has promoted therapeutic limitations against different infections, making research of new molecules that can be applied to treat infections necessary. Antimicrobial peptides (AMPs) are a class of promising antibiotic molecules as they present broad action spectrum, potent activity, and do not easily induce resistance. Several AMPs from scorpion venoms have been described as a potential source for the development of new drugs; however, some limitations to their application are also observed. Here, we describe strategies used in several approaches to optimize scorpion AMPs, addressing their primary sequence, biotechnological potential, and characteristics that should be considered when developing an AMP derived from scorpion venoms. In addition, this review may contribute towards improving the understanding of rationally designing new molecules, targeting functional AMPs that may have a therapeutic application.

Pyrolysis-GCMS of Spirulina platensis: Evaluation of biomasses cultivated under autotrophic and mixotrophic conditions.

Microalgae are autotrophs and CO2 fixers with great potential to produce biofuels in a sustainable way, however the high cost of biomass production is a challenge. Mixotrophic growth of microalgae has been presented as a great alternative to achieve economic sustainability. Thus, the present work reports the energetic characterization of S. platensis biomasses cultivated under autotrophic (A) and mixotrophic conditions using cheese whey waste at different concentrations, 2.5 (M2.5), 5.0 (M5) and 10.0% (M10), in order to analyze the potential production of valuable chemicals and bio-oil by TGA/DTG and Py-GC/MS. The biochemical compositions of the studied biomasses were different due to the influence of different culture mediums. As the whey concentration increased, there was an increase in the carbohydrate content and a decrease in the protein content, which influenced the elemental composition, calorific value, TGA and volatile compounds evaluated by Py-GC/MS at 450°C, 550°C and 650°C. Sample M10 had lower protein content and formed a smaller amount of nitrogenates compounds by pyrolysis at all temperatures evaluated. There was a reduction of 43.8% (450º), 45.6% (550ºC) and 23.8% (650ºC) in the formation of nitrogenates compounds in relation to sample A. Moreover, the temperature also showed a considerable effect in the formation of volatile compounds. The highest yields of nitrogenates compounds, phenols and aromatic and non-aromatic hydrocarbons were observed at 650ºC. The oxygenated, and N and O containing compounds decreased as the temperature increased. Hydrocarbons such as toluene, heptadecane and heneicosane were produced by S.platensis pyrolysis, which makes this biomass attractive for production of high quality bio-oil and valuable chemicals. Therefore, the results showed that it is possible to decrease the formation of nitrogen compounds via manipulation of growth conditions and temperature.

Chemical characteristics of a heteropolysaccharide from Tylopilus ballouii mushroom and its antioxidant and anti-inflammatory activities.

Fucogalactomannan (FGM) is a non-sulphated polysaccharide isolated from the Tylopilus ballouii mushroom. We investigated the chemical characteristics of this FGM using HPLC, chemical methods, and NMR studies ((1)H, (13)C, (1)H/(13)C-HSQC and DEPT-135 spectroscopies) without chemical fragmentation. This polysaccharide consisted primarily of mannose and galactose with variable amounts of fucose and traces of xylose and with MW of 140kDa. Infrared and NMR spectroscopies showed the possible interaction between these polysaccharides and proteins. The antioxidant activity showed for FGM a high inhibition of superoxide and hydroxyl radicals with an IC50 of 1.25 and 1.6mg/mL, respectively. The results of peroxidation tests showed that FGM had an IC50 of 1.72mg/mL. Furthermore, the anti-inflammatory assay showed that FGM reduced edema by 32.8%, 42.0%, and 56% at doses of 30, 50, and 70mg/kg, respectively. Thus, these results suggested a structure and indicated possible anti-inflammatory and antioxidant activities use of these polysaccharides.

Iridoids from leaf extract of Genipa americana

Abstract Genipa americana L., Rubiaceae, is a plant native from Brazil popularly known as "jenipapo". Two iridoids, 1-hydroxy-7-(hydroxymethyl)-1,4aH,5H,7aH-cyclopenta[c]pyran-4-carbaldehyde (1), and iridoid 7-(hydroxymethyl)-1-methoxy-1H,4aH,5H,7aH-cyclopenta[c]pyran-4-carbaldehyde (2) were isolated and identified in the leaf extract of G. americana. Compounds 1 and 2 were identified for the first time in G. americana, and 1 has not been yet described in literature. These substances were analyzed by spectroscopic techniques such as infrared, high resolution mass spectrometry, 1H and 13C 1D; as well as 2D nuclear magnetic resonance. Moreover, the presence of flavonoids was detected by a preliminary analysis by Thin Layer Chromatography.

Antioxidant and anti-inflammatory effect of polysaccharides from Lobophora variegata on zymosan-induced arthritis in rats.

This study analyzes the action of sulfated polysaccharides, fucans, from algae Lobophora variegata on zymosan-induced arthritis in rats. Groups of fucans, obtained after acetone fractionation (0.3-2.0 volumes), were denominated F0.3, F0.5, F0.8, F1, F1.5, and F2. The results that F1 contained a high yield in relation to other fractionated fucans. Chemical and structure analysis of F1 was performed by nuclear magnetic resonance (NMR) and infrared (IR) spectroscopies. The in vitro antioxidant activities of the fraction F1 were also observed. Thus, 2 mg/mL of F1 inhibited the phosphomolybdate in the total antioxidant activity assay. The EC(50) values were 0.3 mg/mL and 0.12 mg/mL for superoxide and hydroxyl radicals, respectively. Fucan F1 (25, 50, and 75 mg/kg by body weight), diclofenac sodium (10 mg/kg), and L-NAME (25 mg/kg) were administered intraperitoneally (i.p.) in rats, according to body weight of different groups of animals (n=6). After 6 h, analyses of cell influx and nitrite levels were conducted. Then after 96 h, analysis of edema and concentration of serum TNF-α was carried out along with histopathological analysis. F1 at 25, 50, and 75 mg/kg i.p. by body weight reduced cell influx in 52.1-96.7% and nitric oxide level in 27.2-39% compared with the control group. The reduction of edema and serum TNF-α was observed at 50 mg/kg i.p. (p<0.001). These results suggest that this heterofucan from the brown algae L. variegata has potential anti-inflammatory activity in acute zymosan-induced arthritis in rats and that antioxidant activity promotes modulation in the cellular redox state.

Glucans from the Caripia montagnei mushroom present anti-inflammatory activity.

Caripia montagnei is a basidiomycete species which contains polysaccharides with immunomodulatory properties. An extract of this mushroom underwent removal of the fat content by organic solvent and subsequently proteolysis. The aqueous phase obtained after proteolysis was precipitated with methanol yielding a fraction containing carbohydrates (98.7+/-3.3%) and protein (1.3+/-0.25%). Chemical analysis, infrared spectroscopy and nuclear magnetic resonance (NMR) showed that the carbohydrate fraction contained (63.3+/-4.1) of beta-glucans and proteins (2.2+/-0.3%). These glucans (50mg/kg of body weight) significantly reduced the inflammatory infiltrate produced by thioglycolate-induced peritonitis by 75.5+/-5.2%, when compared to Wy-14643 (60.3+/-6.1%), PFOA (37.8+/-2.8%) and clofibrate (52.2+/-3.2%), p<0.001, which are of the peroxisome proliferator-activated receptor (PPAR-alpha). L-NAME, a nitric oxide synthase inhibitor, reduced the plantar edema in Wistar rats by 91.4+/-1.3% (p<0.001). A significant reduction in nitric oxide (NO) levels was observed in the exudates when the glucans was used in comparison to carrageenan. The C. montagnei glucans did not present signs of inducing cytotoxicity. A decrease in IL-1ra, IL-10 and IFN-gamma in the peritonitis model was observed. Thus, the results suggest that glucans from the C. montagnei mushroom is an effective immunomodulator and may have potential for anti-inflammatory properties.

Terpenóides e avaliação do potencial antimalárico, larvicida, anti-radicalar e anticolinesterásico de Pouteria venosa (Sapotaceae) / Terpenoids and evaluation of the antimalarial, larvicidal, anti-radicalar and anticholinesterase potential of Pouteria venosa (Sapotaceae)

O presente trabalho descreve o isolamento de quatro triterpenos (taraxerol, ácido ursólico, ácido 3b,19a,23-triidroxiurs-12-en-28-óico e ácido 2a,3a,19a,23-tetraidroxiurs-12-en-28-óico) e um fitoesteróide (espinasterol), bem como a avaliação do potencial antimalárico (cepa NK-65 do Plasmodium berghei), larvicida (larvas do 4º instar do Aedes aegypti), anti-radicalar (2,2-difenil-1-picril-hidrazila, DPPH) e anticolinesterásico de extratos das folhas, cascas do caule e caule de Pouteria venosa (Mart.) Baehni. Todos os compostos isolados estão sendo descritos pela primeira vez nesta espécie e foram identificados com base na análise de dados espectrais (IV e RMN, incluindo APT e DEPT), bem como pela comparação com dados descritos na literatura.

This work describes the isolation of four triterpenes (taraxerol, ursolic acid, 3b,19a,23-trihydroxyurs-12-en-28-oic acid and 2a,3a,19a,23-tetrahydroxyurs-12-en-28-oic acid) and a phytosteroid (spinasterol), as well as a preliminary evaluation of antimalarial (NK-65 strains of Plasmodium berghei), larvicidal (4th instar of Aedes aegypti), anti-radicalar (2,2-diphenyl-1-pycryl-hydrazyl, DPPH) and anticholinesterase activities of Pouteria venosa (Mart.) Baehni extracts from leaves, stem barks and stems. All isolated compounds are being described for the first time in this species and were identified on basis of the spectral data (IR and NMR, including APT, DEPT), as well as by comparison with literature data.