Spasmolytic and antidiarrheal activities of Lippia thymoides (Verbenaceae) essential oil

Lippia thymoides (‘alecrim-do-mato’ or ‘alecrim-do-campo’) is used in Brazilian folk medicine to treat various illnesses, including diarrhea. This work aimed to evaluate in vitro spasmolytic and in vivo antidiarrheal activities of the L. thymoides essential oil (OOS) and to correlate with the traditional use of this plant. In isolated guinea-pig ileum, OOS presented a concentration-dependent spasmolytic activity in preparations pre-contracted with KCl 40 mM [EC₅₀ = 16.89 (11.56–24.66) μg/mL], and antagonized phasic contractions induced by 1 μM carbachol [IC₅₀ = 42.71 (37.35–48.83) μg/mL] or histamine [IC₅₀ = 32.38 (27.44–38.20) μg/mL]. In mice, OOS at 400 mg/kg reduced intestinal transit, at 200 and 400 mg/kg reduced total stool mass and at 400 mg/kg reduced intestinal fluid accumulation. It was shown that the antidiarrheal effect of OOS is related to the inhibition of smooth muscle contraction and may be due to the presence of major compound β-caryophyllene in this essential oil.     

Potential of Carvacrol and Thymol in Reducing Biofilm Formation on Technical Surfaces

Polyvinyl chloride (PVC), polypropylene (PP), polyethylene (PE), and stainless steel (SS) are commonly used in medicine and food production technologies. During contact with microorganisms on the surface of these materials, a microbial biofilm is formed. The biofilm structure is difficult to remove and promotes the development of pathogenic bacteria. For this reason, the inhibition of biofilm formation in medical and food production environments is very important. For this purpose, five naturally occurring compounds were used for antimicrobial screening tests. The two with the best antimicrobial properties were chosen to inhibit the biofilm formation of Staphylococcus aureus and Pseudomonas aeruginosa. After 3 days of exposure, thymol reduced the amount of biofilm of Pseudomonas aeruginosa within the range of 70–77% and 52–75% for Staphylococcus aureus. Carvacrol inhibited the formation of biofilms by up to 74–88% for Pseudomonas aeruginosa and up to 86–100% for Staphylococcus aureus. Those phenols decreased the enzyme activity of the biofilm by up to 40–100%. After 10 days of exposure to thymol, biofilm formation was reduced by 80–100% for Pseudomonas aeruginosa and by about 79–100% for Staphylococcus aureus. Carvacrol reduced the amount of biofilm by up to 91–100% for Pseudomonas aeruginosa and up to 95–100% for Staphylococcus aureus.     

Changes in the Leaf Surface Chemistry of Apium graveolens (Apiaceae) stimulated by jasmonic acid and perceived by a specialist insect

Treatment of celery leaves (Apium graveolens, cv. secalinum) with Jasmonic acid (JA, 1 ) or analogues of amino-acid conjugates of Jasmonic acid such as the leucine conjugate with 1-oxoindan-4-carboxylic acid (IN-ILE, 2 ) stimulated the biosynthesis of the furanocoumarins psoralen ( 6 ), xanthotoxin ( 8 ), bergapten ( 7 ), and isopimpinellin ( 9 ). Besides the increase of the compounds within the leaf, a significant amount (ca. 20%) of the total furanocoumarins was deposited on the surface of the leaf. The two monomethoxy furanocoumarins, 7 and 8 , began to increase steadily and simultaneously within the leaf and on the leaf surface ca. 40 h after the onset of the jasmonic-acid stimulus. Within the leaf, the ratio 7/8 was ca. 1.3:1.0, while among the surface lipids 8 dominated ( 7/8 0.8:1.0), indicating that the export of the compounds to the surface is not a simple diffusive translocation along the oil-ducts. Females of the carrot fly (Diptera, Psilidae) responded with an increased oviposition to the altered leaf surface chemistry of the JA-treated celery plants. The effect shown for total leaves was corroborated by surrogate leaves treated with leaf-surface extracts of JA-induced leaves. Based on the known stimulation of oviposition by furanocoumarins, we conclude that the enhanced amount of furanocoumarins on the surface can explain the insects' preference for the JA-stimulated plants. This is the first report of a JA-induced change of the surface chemistry of a plant and an increase of the acceptability of treated leaves for a specialist insect, like the carrot fly.     

Allelopathic effect of the ethanol extract and fractions of the aerial parts of Lippia alba (Verbenaceae)

Lippia alba, belonging to the Verbenaceae family, is one of the most commonly utilized medicinal plants in folk medicine. The allelopathic activity was assessed using seeds of Lactuca sativa (lettuce) and Allium cepa (onion) by assessing the growth of the radicle and hypocotyl. The tests showed allelopathic efficiency in inhibiting the growth of lettuce and onion seeds. The best results for allelopathic activity were presented by the dichloromethane (DCM) fraction of the fresh plant, which inhibited radicle (23.04–100% lettuce and 64.17–66.36% onion) and hypocotyl (16.77–100% lettuce and 65.10–69.43% onion) formation, and as well as the DCM fraction of the dry plant, which also inhibited radicle (30.74–82.83% lettuce and 63.50–93.67% onion) and hypocotyl (24.12–70% lettuce and 69.07–79.95% onion) formation. Based on these results, it was found that the aerial parts of L. alba are rich in bioactive substances, suggesting the possibility of using of L. alba as a natural herbicide.     

Phytochemical characterisation and bioprospection for antibacterial and antioxidant activities of Lippia alba Brown ex Britton & Wilson (Verbenaceae)

Ethanol extract and fractions obtained from fresh and dry aerial parts of Lippia alba were examined in order to determine their phytochemical composition, antioxidant capacity and antibacterial activities. The ethanol extracts and fractions exhibited an antioxidant effect by the DPPH assay, especially samples of fresh plant. HPLC analysis of the ethyl acetate fractions identified the presence of phenolic acids and flavonoids. The ethanol extract and fractions showed activity against reference and multidrug-resistant strains of Staphylococcus aureus and Enterococcus faecalis (MIC range 2000–250 μg/mL). The hexane and dichloromethane fractions of fresh plant showed better activity against reference strains of Escherichia coli (MIC of 250 and 125 μg/mL, respectively), but all extracts and fractions were less active against multidrug-resistant strains of all the Gram-negative species evaluated. The results showed that the extract and fractions of L alba aerial parts showed antibacterial activity, even against multidrug-resistant Gram-positive bacteria, and antioxidant effect (DPPH assay).     

Identification of the major compounds in extracts ofVerbena officinalis L. (Verbenaceae) by HPLC with post-column derivatization

Summary A qualitative reversed-phase HPLC method has been developed for the analysis of 50% MeOH extracts ofVerbena officinalis L. (Verbenaceae) leaves. The method enables separation of the main constituents: iridoids, flavonoids and phenolic acid derivatives. Simultaneous detection at different wavelengths, measurement of the UV spectrum of each separated compound during elution and co-injection of reference substances facilitated easy and rapid identification of verbenalin, hastatoside and verbascoside. As some of these compounds, mainly flavonoids, have closely related structures, however, characterization by derivatization with reagents inducing a shift of UV absorption maxima was required. This furnished additional structural information. The reagents were adapted for compatibility with the solvent system used for the chromatographic separation.     

In Silico Evaluation of the Antimicrobial Activity of Thymol—Major Compounds in the Essential Oil of Lippia thymoides Mart. & Schauer (Verbenaceae)

In this paper, we evaluated the drug-receptor interactions responsible for the antimicrobial activity of thymol, the major compound present in the essential oil (EO) of Lippia thymoides (L. thymoides) Mart. & Schauer (Verbenaceae). It was previously reported that this EO exhibits antimicrobial activity against Candida albicans (C. albicans), Staphylococcus aureus (S. aureus), and Escherichia coli (E. coli). Therefore, we used molecular docking, molecular dynamics simulations, and free energy calculations to investigate the interaction of thymol with pharmacological receptors of interest to combat these pathogens. We found that thymol interacted favorably with the active sites of the microorganisms’ molecular targets. MolDock Score results for systems formed with CYP51 (C. albicans), Dihydrofolate reductase (S. aureus), and Dihydropteroate synthase (E. coli) were −77.85, −67.53, and −60.88, respectively. Throughout the duration of the MD simulations, thymol continued interacting with the binding pocket of the molecular target of each microorganism. The van der Waals (ΔE_vdW = −24.88, −26.44, −21.71 kcal/mol, respectively) and electrostatic interaction energies (ΔE_ele = −3.94, −11.07, −12.43 kcal/mol, respectively) and the nonpolar solvation energies (ΔG_NP = −3.37, −3.25, −2.93 kcal/mol, respectively) were mainly responsible for the formation of complexes with CYP51 (C. albicans), Dihydrofolate reductase (S. aureus), and Dihydropteroate synthase (E. coli).     

Effect of urea on poly(methacrylic acid) and poly(acrylic acid) aqueous solutions

The effects of urea on aqueous solutions of both poly(methacrylic acid (PMA) and poly(acrylic acid) (PAA) have been investigated by using potentiometry, viscometry and study of the fluorescence of Auramine O, a cationic dye. The viscosity behaviour of unionized PMA obtained from direct dissolution of solid powder shows that the unneutralized macromolecules can be associated in water. The stability of such “aggregates” seems weak as indicated by their disappearance as soon as the charge density is very low. For PMA salt solution percolated through a cation (H⁺) exchange resin column, no association is observed. The pH-dependent conformational behaviour of PMA which, contrary to PAA, presents compact conformations in water at low charge density is discussed in terms of solvophobic/solvophilic interactions. It is shown that, even for urea concentration up to 8 M, the compact conformations of PMA are not completely destroyed. The formation of H⁺/urea complex is taken into account.     

Bioimpact of application of pesticides with plant growth hormone (gibberellic acid) on target and non-target microorganisms

The objective of this investigation was to determine the impacts of fungicide, insecticide, plant growth hormone (gibberellic acid) on soil microbiota, and the growth characteristics of Aspergillus flavus. In the fungicide or insecticide mixed with plant growth hormone treated soil sample, the total viable number of soil microbiota was found to be higher than that of the soil treated with fungicide or insecticide alone. Moderate effect of insecticide used on the total number of fungi was observed. On the other hand the effect of insecticide on soil bacteria was more than effect of fungicide, and the negative effect of fungicide on soil bacteria was observed particularly at latent periods (15 and 20 days) of application. A great sensitivity to fungicide and insecticide was observed in the case of nitrogen fixing bacteria. At 15 days after fungicide and insecticide application the adverse effect was found. Morphological deformations were clear in A. flavus cultivated on medium containing fungicide, the fungus failed to form conidiospores, conidiophores and vesicles. Intermediate and terminal outgrowths like blisters and terminal vesicle originate from hyphae. The addition of plant growth hormone reduced the effect of fungicide on fungus.     

Effect of biodegradation conditions on morphology of ternary compositions of low density polyethylene with poly(lactic acid) and starch

Polyethylene–poly(lactic acid)–starch ternary compositions were obtained in a rotor disperser under conditions of shear deformation. Their biodegradation under the action of mold fungus spores and soil was investigated using IR spectroscopy and SEM. The changes under the fungal action revealed that the ternary blends represented a nutrient medium, while the exposure to soil led to alteration in morphology due to the whole range of numerous environmental factors.     

Increased synthesis of a new oleanane-type saponin in hairy roots of marigold (Calendula officinalis) after treatment with jasmonic acid

Native plant of marigold (Calendula officinalis L.) synthesizes oleanolic acid saponins classified as glucosides or glucuronides according to the first residue in sugar chain bound to C-3 hydroxyl group. Hairy root culture, obtained by transformation with Agrobacterium rhizogenes strain 15834, exhibit a potent ability of synthesis of oleanolic acid glycosides. The HPLC profile of saponin fraction obtained from C. officinalis hairy roots treated with plant stress hormone, jasmonic acid, showed the 10-times increase of the content of one particular compound, determined by NMR and MALDI TOF as a new bisdesmoside saponin, 3-O-β-d-glucuronopyranosyl-28-O-β-d-galactopyranosyl-oleanolic acid. Such a diglycoside does not occur in native C. officinalis plant. It is a glucuronide, whereas in the native plant glucuronides are mainly accumulated in flowers, while glucosides are the most abundant saponins in roots. Thus, our results revealed that the pathways of saponin biosynthesis, particularly reactions of glycosylation, are altered in C. officinalis hairy root culture.     

Characterization of an antimicrobial poly(lactic acid) film prepared with poly(ε‐caprolactone) and thymol for active packaging

Antimicrobial active films based on poly(lactic acid) (PLA) were prepared with poly(ε‐caprolactone) (PCL) and thymol (0, 3, 6, 9, and 12 wt%) by solvent casting methods. The films were characterized by thermal, structural, mechanical, gas barrier, and antimicrobial properties. Scanning electron microscopy analysis revealed that the surface of film became rougher with certain porosity when thymol was incorporated into the PLA/PCL blends. Thymol acted as plasticizers, which reduce the intermolecular forces of polymer chains, thus improving the flexibility and extensibility of the films. The addition of PCL into the pure PLA film decreased the glass transition temperature of the films. The presence of thymol decreased the crystallinity of PLA phase, but did not affect the thermal stability of films. Water vapor barrier properties of films slightly decreased with the increase of thymol loading. The antimicrobial properties of thymol containing films showed a significant activity against Escherichia coli and Listeria monocytogenes. The results indicated the potential of PLA/PCL/thymol composites for applications in antimicrobial packaging. Copyright © 2014 John Wiley & Sons, Ltd.     

Enhanced daidzin production from jasmonic and acetyl salicylic acid elicited hairy root cultures of Psoralea corylifolia L. (Fabaceae)

Daidzin (7-O-glucoside of daidzein) has several pharmacological benefits in herbal remedy, as antioxidant and shown antidipsotropic activity. Hairy root culture of Psoralea corylifolia L. was developed for biomass and enhanced daidzin production using signalling compounds such as jasmonic acid (JA) and acetyl salicylic acid (ASA). Best response of 2.8-fold daidzin (5.09% DW) with 1 μM JA treatment after second week and 7.3-fold (3.43% DW) with 10 μM JA elicitation after 10th week was obtained from hairy roots compared to untreated control. ASA at 10 μM promoted 1.7-fold increase in daidzin (1.49% DW) content after seventh week compared to control (0.83% DW). Addition of 25 μM ASA resulted in 1.44% DW daidzin (1.5-fold increase) with 0.91% DW in control after fifth week and 1.44% DW daidzin (2.3-fold increase) after eighth week when compared to untreated control (0.62% DW). Reduced biomass with increased daidzin content was facilitated by elicited hairy root cultures.     

Effect of pH on Poly(acrylic acid) Solution Polymerization

The free-radical redox-initiated aqueous solution polymerization of fully and partially neutralized acrylic acid was carried out at room temperature under full exposure to air. The effect of neutralization degree on the polymerization rate and product properties was studied. Increasing neutralization of the reaction mixture with sodium hydroxide resulted in greater conversion of acrylic acid to sodium acrylate. The rate of polymerization, determined from a gravimetric off-line water removal technique, was shown to decrease significantly with decreasing degree of neutralization. Molecular weight also decreased with decreasing degree of neutralization. The glass transition temperature and hydrophilicity of the polymer product decreased with increasing degree of neutralization. In-line infrared monitoring was also used to monitor the reaction progress and was shown to be an effective tool for this purpose.     

Effect of experimental conditions on nano-indentation response of low density polyethylene (LDPE)

Nano-indentation is an interesting tool for analyzing nano-scale mechanical properties. The analysis of nano-mechanical properties as a function of experimental conditions is very critical for designing engineering components. In this study, nano-indentation experiments were performed by considering different values of amplitude (1, 5, 10?nm), frequency (11.2, 22.5, 45?Hz), strain rate (0.02, 0.05, 0.1, 0.2, 1?s^?1), peak load (10, 30, 100?mN) and hold time (1, 3, 5, 10, 20, 50, 100?sec) to analyze their effect on the mechanical properties of LDPE. The results showed that the effect of amplitude and frequency on the nano-mechanical properties of LDPE were negligible. Load-displacement curves displayed a shift towards higher indentation depths along with a decrease in peak load from 20.6 to 14.8?mN by having a decrease in strain rate from 1 to 0.02?s^?1. Elastic modulus and hardness values exhibited a decrease with an increase in hold time. Logarithmic creep model was used to fit the experimental data of creep as a function of holding time which showed good agreement (r² ≥ 0.97) with the experimental values. Recommended holding times are also suggested to eliminate the creep and nose problem in order to achieve high accuracy in measurements.     

Effect of sepiolite on the biodegradation of poly(lactic acid) and polycaprolactone

PLA and PCL nanocomposites prepared by adding 5 wt% of a sepiolite (SEPS9) were degraded in compost, leading to effective degradation for all samples.PLA and PLA/SEPS9 seem to be mainly degraded by a bulk mechanism, showing a significant level of polymer degradation, however the presence of SEPS9 particles partially delays the degradation probably due to a preventing effect of these particles on polymer chain mobility and/or PLA/enzymes miscibility. PCL and PCL/SEPS9 showed a preferential surface mechanism of degradation; and in contrast to PLA, sepiolite does not present a considerable barrier effect on the degradation of PCL.     

Influence of charge density on the swelling of colloidal poly(N-isopropylacrylamide-co-acrylic acid) microgels

The volume phase transition of colloidal poly(N-isopropylacrylamide-co-acrylic acid) microgels depends in a complex way on the effective charge density within the polymer network. A series of monodisperse PNIPAM/AAc microgels with different content of acrylic acid were synthesized by surfactant-free emulsion polymerization employing sonication instead of a conventional stirring technique. Subsequently, the colloids were characterized by dynamic light scattering and electron microscopy. Potentiometric titrations provided the amount of carboxyl groups incorporated into the copolymer. The effective charge density was systematically controlled by the content of acrylic acid monomers, the pH value of the suspension, and the salt concentration. The hydrodynamic dimensions of the microgels have been measured by dynamic light scattering. The swelling/deswelling behavior is determined by the delicate balance between hydrophobic attraction of NIPAM and the repulsive electrostatic interactions of the carboxylate group of the acrylic acid moieties. Compared to their macroscopic counterparts the charged microgel particles show a significantly different swelling/deswelling behavior. This manifests in the occurrence of a two-step volume phase-transition process with increasing acrylic acid content. Hydrogen bonding has to be considered to understand this two step volume phase transition uniquely observed for colloidal microgels. Another interesting phenomenon presented here is the reversible formation of well-defined aggregates at low pH and under high salt conditions.     

Effect of pressure on dissociation of poly(methacrylic acid) in aqueous solution

The effect of pressure (up to 6300 atm) on the electrical conductivity of poly(methacrylic acid) (PMA) in aqueous solution was investigated in the concentration range 0.001M to 0.1M (in equivalents of carboxyl groups) at 30°C. The degree of ionization α calculated from the conductivity data increased linearly with pressure. From dilatometric measurements of volume changes when PMA was partially neutralized with NaOH, a conformational transition was confirmed to take place above α = 0.12, and the volume change accompanying the dissociation of the side chain groups (carboxyl groups) was found to be ? 12.5 ml/H⁺. It may be inferred from the values of α at each polymer concentration and from the dilatometric data that there is no conformational transition from a compact structure to the coiled form below at least 6,000 atm in the absence of Na⁺ counterions.     

Biodegradable Poly(butylene adipate-co-terephthalate)/Poly(glycolic acid) Films: Effect of Poly(glycolic acid) Crystal on Mechanical and Barrier Properties

Binary biodegradable polymers films, poly(butylene adipate-co-terephthalate)(PBAT) and poly(glycolic acid)(PGA), were prepared through batch melt mixing to obtain Film Ⅰ and Film Ⅱ under two different processing conditions. PGA crystals played a major role in enhancing the mechanical and barrier properties of the films. For Film Ⅰ, there were initial PGA crystals before the film blowing process, the PGA molecular chain further crystallized, forming the oriented crystallization of PGA. Moreover, ...     

Effect of dyeing on melting behavior of poly(lactic acid) fabric

The effect of the dyeing on the melting behavior of poly(lactic acid) fabrics was investigated by differential scanning calorimeter. The DSC melting peaks at 10°C min^-1 of the untreated poly(lactic acid) fabric were observed at a temperature higher than those of the dyed fabrics. The restricting force from the extended tie molecules along the fiber axis seems to decrease in the dyeing process. When the sample was rapidly heated, the crystallites melted at lower temperatures since recrystallization was restricted. It was estimated, based on the heating-rate dependency of melting behavior, that the original crystallites of the untreated sample melted at 146.1°C and those of the dyed samples melted at higher temperatures, suggesting that their crystallites are grown to be more perfect in the dyeing process. This revised version was published online in July 2006 with corrections to the Cover Date.