Encapsulation of eucalyptus and Litsea cubeba essential oils using zein nanopolymer: Preparation, characterization, storage stability, and antifungal evaluation.

The encapsulation of essential oils (EOs) in protein-based biopolymeric matrices stabilized with surfactant ensures protection and physical stability of the EO against unfavorable environmental conditions. Accordingly, this study prepared zein nanoparticles loaded with eucalyptus essential oil (Z-EEO) and Litsea cubeba essential oil (Z-LEO), stable and with antifungal activity against Colletotrichum lindemuthianum, responsible for substantial damage to bean crops. The nanoparticles were prepared by nanoprecipitation with the aid of ultrasound treatment and characterized. The nanoparticles exhibited a hydrodynamic diameter close to 200 nm and PDI < 0.3 for 120 days, demonstrating the physical stability of the carrier system. Scanning electron microscopy and Transmission electron microscopy revealed that the nanoparticles were smooth and uniformly distributed spheres. Fourier-transform infrared spectroscopy showed interaction between zein and EOs through hydrogen bonding and hydrophobic interactions. Thermogravimetric analysis demonstrated the thermal stability of the nanoparticles compared to pure bioactive compounds. The nanoparticles exhibited a dose-dependent effect in inhibiting the fungus in in vitro testing, with Z-EEO standing out by inhibiting 70.0 % of the mycelial growth of C. lindemuthianum. Therefore, the results showed that zein has great potential to encapsulate hydrophobic compounds, improving the applicability of the bioactive compound as a biofungicide, providing protection for the EO.

Inclusion Complexes of Litsea cubeba (Lour.) Pers Essential Oil into ß-Cyclodextrin: Preparation, Physicochemical Characterization, Cytotoxicity and Antifungal Activity.

The uses of natural compounds, such as essential oils (EOs), are limited due to their instability to light, oxygen and temperature, factors that affect their application. Therefore, improving stability becomes necessary. The objective of this study was to prepare inclusion complexes of Litsea cubeba essential oil (LCEO) with ß-cyclodextrin (ß-CD) using physical mixing (PM), kneading (KN) and co-precipitation (CP) methods and to evaluate the efficiency of the complexes and their physicochemical properties using ATR-FTIR, FT-Raman, DSC and TG. The study also assessed cytotoxicity against human colorectal and cervical cancer cells and antifungal activity against Aspergillus flavus and Fusarium verticillioides. The complexation efficiency results presented significant evidence of LCEO:ß-CD inclusion complex formation, with KN (83%) and CP (73%) being the best methods used in this study. All tested LCEO:ß-CD inclusion complexes exhibited toxicity to HT-29 cells. Although the cytotoxic effect was less pronounced in HeLa tumor cells, LCEO-KN was more active against Hela than non-tumor cells. LCEO-KN and LCEO-CP inclusion complexes were efficient against both toxigenic fungi, A. flavus and F. verticillioides. Therefore, the molecular inclusion of LCEO into ß-CD was successful, as well as the preliminary biological results, evidencing that the ß-CD inclusion process may be a viable alternative to facilitate and increase future applications of this EO as therapeutic medication, food additive and natural antifungal agent.

Litsea glaucescens Kuth possesses bactericidal activity against Listeria monocytogenes.

Background: Litsea glaucencens Kuth is an aromatic plant used for food seasoning food and in Mexican traditional medicine. Among, L. glaucencens leaves properties, it has proven antibacterial activity which can be used against opportunistic pathogens like Listeria monocytogenes, a foodborne bacteria that is the causal agent of listeriosis, a disease that can be fatal in susceptible individuals. The aim of this work was to investigate the antibacterial activity of L. glaucescens Kuth leaf extracts against L. monocytogenes and to identify its bioactive components. Material and Methods: L. glaucences leaves were macerated with four solvents of different polarity (n-hexane, dichloromethane, ethyl acetate, and methanol). To determine the capacity to inhibit bacterial proliferation in vitro, agar diffusion and microdilution methods were used. Next, we determined the minimal bactericidal concentration (MBC). Finally, we determined the ratio of MBC/MIC. Metabolites present in the active methanolic extract from L. glaucescens Kuth (LgMeOH) were purified by normal-phase open column chromatography. The structure of the antibacterial metabolite was determined using nuclear magnetic resonance (1H, 13C, COSY, HSQC) and by comparison with known compounds. Results: The LgMeOH extract was used to purify the compound responsible for the observed antimicrobial activity. This compound was identified as 5,7-dihydroxyflavanone (pinocembrin) by analysis of its spectroscopic data and comparison with those described. The MIC and MBC values obtained for pinocembrin were 0.68 mg/mL, and the ratio MBC/MIC for both LgMeOH and pinocembrin was one, which indicates bactericidal activity. Conclusion: L. glaucences Kuth leaves and its metabolite pinocembrin can be used to treat listeriosis due the bactericidal activity against L. monocytogenes.

Acaricidal activity of the essential oils from Leptospermum scoparium, Origanum vulgare and Litsea cubeba on Rhipicephalus microplus: Influence of the solvents and search for fractions with higher bioactivity.

The use of natural products in research on tick control for Rhipicephalus microplus is increasing year by year, with promising results. In this regard, the aim of the present study was to phytochemically characterize the essential oils (EOs) of Leptospermum scoparium, Origanum vulgare and Litsea cubeba, and to evaluate the acaricidal activity of these EOs in solutions prepared using ethanol, dimethylsulfoxide (DMSO) and Tween 80 on larvae and females of R. microplus. In addition, three L. scoparium fractions were also isolated and their acaricidal activity on these larvae and adult females was tested. Gas chromatography and mass spectrometry results showed that cis-calamenene (29.82 %), carvacrol (64.85 %) and geranial (42.44 %) were the majority compounds of L. scoparium, O. vulgare and L. cubeba, respectively. Three fractions were isolated from L. scoparium: A1, rich in sesquiterpene hydrocarbons, and A2 and A3, rich in ß-triketones. Bioassays on unfed larvae (immersion test) were performed using all the EOs at concentrations from 2.5 to 10.0 mg/mL; and using the three fractions obtained from L. scoparium EO at concentrations from 0.625 to 10 mg/mL. We observed 100 % mortality of larvae in all treatments with L. scoparium EO at all concentrations (diluted both in DMSO and in ethanol), and in treatments with O. vulgare EO diluted in DMSO. However, L. cubeba EO only gave rise to more than 99 % mortality at a concentration of 10 mg/mL, using the same solvents. For engorged females, the immersion test was performed at concentrations from 2.5 to 10.0 mg/mL. Percentage control greater than 90 % was observed only at the highest concentrations of L. scoparium and O. vulgare EOs diluted in DMSO and ethanol, while L. cubeba EO did not reach 90 % control in any of the treatments. In tests on L. scoparium fractions, larval mortality in the fractions rich in ß-triketones (A2 and A3) was above 97 % at a concentration of 2.5 mg/mL, while in the A1 fraction, rich in sesquiterpene hydrocarbons, at the same concentration (2.5 mg/mL), mortality did not reach 22 %. In the adult immersion test, the percentage control was higher than 98 % at the lowest concentration (2.5 mg/mL) of the A1 fraction, while in the treatments with the fractions A2 and A3, the control levels were 16 and 50 %, respectively. Thus, we can conclude that the EOs of L. scoparium, O. vulgare and L. cubeba have acaricidal activity on R. microplus, as also do the fractions derived from L. scoparium EO.

Plant-derived bioactive compounds produced by Streptomyces variabilis LCP18 associated with Litsea cubeba (Lour.) Pers as potential target to combat human pathogenic bacteria and human cancer cell lines.

To date, endophytic actinomycetes have been well-documented as great producers of novel antibiotics and important pharmaceutical leads. The present study aimed to evaluate potent bioactivities of metabolites synthesized by the strain LCP18 residing in the Vietnamese medicinal plant Litsea cubeba (Lour.) Pers towards human pathogenic bacteria and human cancer cell lines. Endophytic actinomycete strain LCP18 showed considerable inhibition against seven bacterial pathogens and three human tumor cell lines and was identified as species Streptomyces variabilis. Strain S. variabilis LCP18 was phenotypically resistant to fosfomycin, trimethoprim-sulfamethoxazole, dalacin, cefoxitin, rifampicin, and fusidic acid and harbored the two antibiotic biosynthetic genes such as PKS-II and NRPS. Further purification and structural elucidation of metabolites from the LCP18 extract revealed five plant-derived bioactive compounds including isopcrunetin, genistein, daidzein, syringic acid, and daucosterol. Among those, isoprunetin, genistein, and daidzein exhibited antibacterial activity against Salmonella typhimurium ATCC 14,028 and methicillin-resistant Staphylococcus epidermidis ATCC 35,984 with the MIC values ranging from 16 to 128 µg/ml. These plant-derived compounds also exhibited cytotoxic effects against human lung cancer cell line A549 with IC50 values of less than 46 µM. These findings indicated that endophytic S. variabilis LCP18 can be an alternative producer of plant-derived compounds which significantly show potential applications in combating bacterial infections and inhibition against lung cancer cell lines.

Litsea cubeba essential oil: chemical profile, antioxidant activity, cytotoxicity, effect against Fusarium verticillioides and fumonisins production.

The purpose of this study was to determine the chemical profile of Litsea cubeba essential oil, carry out an in vitro evaluation of its antioxidant potential and its cytotoxicity, as well as its antifungal and antimicotoxigenic activities against Fusarium verticillioides. Most of the compounds observed in the EO were neral (32.75%) and geranial (37.67%). The radical scavenging capacity of 2,2-diphenyl-1-picrylhydrazyl and 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid was 104.4 and 56.4 mmol Trolox mg-1, respectively, indicating good antioxidant activity. The EO studied by us revealed cytotoxic effect against HT-29 and HeLa cancer cells. The Minimum Inhibitory and Minimum Fungicidal Concentrations against F. verticillioides were both 125 µg mL-1. Morphological investigation, performed by fluorescence microscopy and scanning electron microscopy, showed that hyphae and microconidia structures underwent changes after treatment with the EO. Analyses performed with the EO strongly reduced the mycelial development of F. verticillioides and the synthesis of fumonisins B1 and B2 in dose-dependence effect compared (P < 0.01) with the fungal control (105 conidia mL-1) and positive control (fludioxonil + metalaxyl-M). Thus, the results obtained in vitro suggest that L. cubeba EO has excellent antioxidant, fungicidal, and antimycotoxigenic effects.

Sex differential marker FD for rapid sex identification of Litsea cubeba.

Litsea cubeba is an important economic tree in China. Sex identification of the species is required to reduce breeding costs. Molecular biology is an ideal method to achieve this aim because of the lack of morphological differences between male and female plants, especially at the seedling stage. Sequence-related amplified polymorphism was used to amplify sex-related bands. Following sequencing, the amplified fragment Dwas used to create a sequence-characterized amplified region (SCAR) marker, FD. The SCAR marker is approximately 750 bp, is female-specific, and is expected to be useful for L. cubeba breeding programs. Furthermore, the amplified fragment L had homology to sex-determining-related genes of other species. Quantitative real-time polymerase chain reaction analysis of this fragment during flower bud development identified expression differences between male and female plants.

In vivo analgesic, antipyretic, and anti-inflammatory potential in Swiss albino mice and in vitro thrombolytic activity of hydroalcoholic extract from Litsea glutinosa leaves.

BACKGROUND: The study was conducted to evaluate the in vitro thrombolytic activity, and in vivo analgesic, anti-inflammatory and antipyretic potentials of different hydrocarbon soluble extracts of Litsea glutinosa leaves for the first time widely used in the folkloric treatments in Bangladesh. This work aimed to create new insights on the fundamental mechanisms of the plant extracts involved in these activities. RESULTS: In thrombolytic activity assay, a significant clot disruption was observed at dose of 1 mg/mL for each of the extracts (volume 100 µL) when compared to the standard drug streptokinase. The n-hexane, ethyl acetate, chloroform, and crude methanolic extracts showed 32.23 ± 0.26, 37.67 ± 1.31, 43.13 ± 0.85, and 46.78 ± 0.9% clot lysis, respectively, whereas the positive control streptokinase showed 93.35 ± 0.35% disruption at the dose of 30,000 I.U. In hot plate method, the highest pain inhibitory activity was found at a dose of 500 mg/kg of crude extract (15.54 ± 0.37 sec) which differed significantly (P <0.01 and P <0.001) with that of the standard drug ketorolac (16.38 ± 0.27 sec). In acetic acid induced writhing test, the crude methanolic extract showed significant (P <0.01 and P <0.001) analgesic potential at doses 250 and 500 mg/kg body weight (45.98 and 56.32% inhibition, respectively), where ketorolac showed 64.36% inhibition. In anti-inflammatory activity test, the crude methanolic extract showed significant (P <0.001) potential at doses 250 and 500 mg/kg body weight (1.51 ± 0.04 and 1.47 ± 0.03 mm paw edema, respectively), where ketorolac showed 1.64 ± 0.05 mm edema after 3 h of carrageenan injection. In antipyretic activity assay, the crude extract showed notable reduction in body temperature (32.78 ± 0.46°C) at dose of 500 mg/kg-body weight, when the standard (at dose 150 mg/kg-body weight) exerted 33.32 ± 0.67°C temperature after 3 h of administration. CONCLUSIONS: Our results yield that the crude hydroalcoholic extract has better effects than the other in all trials. In the context, it can be said that the leaves of L. glutinosa possess remarkable pharmacological effects, and justify its traditional use as analgesic, antipyretic, anti-inflammatory, and thrombolytic agent.

Efecto de la concentración de saponinas en la actividad hemolítica de extractos de ocho plantas de uso medicinal en Guatemala / Efecto de la concentración de saponinas en la actividad hemolítica de extractos de ocho plantas de uso medicinal en Guatemala

Las plantas se han utilizado desde tiempos antiguos para el tratamiento de todo tipo de afecciones, considerándose en el tiempo actual como una alternativa para la medicina tradicional natural que está fuera del alcance de la población en países como el nuestro. La presente investigación tuvo como principal objetivo determinar la actividad hemolítica in vitro de ocho plantas de uso medicinal en Guatemala (Byrsonima crassifolia, Bixa orellana, Lippia graveolens, Litsea guatemalensis, Petiveria alliacea, Phlebodium pseudoaureum, Smilax domingensis, Solanum nigrescens). Estudios previos han demostrado que estas plantas contienen saponinas, las cuales son un grupo de glucósidos solubles en agua y que contienen características tensoactiva, hemolíticas y citotóxicas (Bruneton, 2001). Se comprobó la presencia de saponinas en las ocho plantas estudiadas, por medio de pruebas cualitativas iniciando con la prueba de espuma. Posteriormente se realizó la identificación por medio de análisis cromatográfico, comparándose con estándares de saponinas esteroidales. La cuantificación de las mismas se realizó por medio de espectrofotometría obteniéndose para Byrsonima crassifolia 1.78%±0.222%, Bixa orellana 0.80%±0.04%, Lippia graveolens 0.21%±0.06%, Litsea guatemalensis 3.19%±0.10%, Petiveria alliacea 0.92%±0.04%, Phlebodium pseudoaureum 1.06%±0.05%, Smilax domingensis 0.21%±0.006% y Solanum nigrescens 1.42%±0.007%. La actividad hemolítica de los extractos de las plantas determinada cualitativamente en agar sangre, no se presentó en ninguno de los extractos estudiados, al compararla con un estándar de saponinas que presentó un halo de hemólisis de 4mm de diámetro. Así mismo se realizó otro ensayo cualitativo en tubo, el cual demostró la presencia de hemólisis en cinco de las ocho plantas de estudio, la cuales son Bixa orellana, Byrsonima crassifolia, Petiveria alliacea, Phlebodium pseudoaureum y Smilax domingensis La cuantificación de la hemólisis de los extractos en estudio, se realizó por medio de espectrofotometría a dos diferentes temperaturas, 25°C y 37°C y a diferentes concentraciones, en donde se obtuvo un mayor porcentaje de actividad hemolítica con los extractos de Solanum nigrescens, Petiveria alliacea y Bixa orellana. También se demostró que la hemólisis aumenta mientras aumenta la concentración del extracto. Se calculó para cada extracto la concentración a la cual ocurre el 50% de hemólisis (CH50), para establecer diferencias de hemólisis a dos temperaturas: 25°C y 37°C obteniendo una correlación entre la concentración de los extractos y la actividad hemolítica. A 25°C solamente Bixa orellana, Petiveria alliacea y Solanum nigrescens tienen importante actividad hemolítica; mientras que a 37°C tienen improtancia Bixa orellana, Litsea guatemalensis, Petiveria alliacea y Solanum nigrescens. En este estudio, la actividad hemolítica de las saponinas de las plantas pudo ser comprobada, sin embargo no ha sido específica por lo que es importante profundizar en el tema dentro de los límites de la lisis y la toxicidad a los eritrocitos, ya que son plantas de uso popular medicinal.

In vivo analgesic, antipyretic, and anti-inflammatory potential in Swiss albino mice and in vitro thrombolytic activity of hydroalcoholic extract from Litsea glutinosaleaves

BACKGROUND: The study was conducted to evaluate the in vitro thrombolytic activity, and in vivo analgesic, anti-inflammatory and antipyretic potentials of different hydrocarbon soluble extracts of Litsea glutinosaleaves for the first time widely used in the folkloric treatments in Bangladesh. This work aimed to create new insights on the fundamental mechanisms of the plant extracts involved in these activities. RESULTS: In thrombolytic activity assay, a significant clot disruption was observed at dose of 1 mg/mL for each of the extracts (volume 100 µL) when compared to the standard drug streptokinase. The n-hexane, ethyl acetate, chloroform, and crude methanolic extracts showed 32.23 ± 0.26, 37.67 ± 1.31, 43.13 ± 0.85, and 46.78 ± 0.9% clot lysis, respectively, whereas the positive control streptokinase showed 93.35 ± 0.35% disruption at the dose of 30,000 I.U. In hot plate method, the highest pain inhibitory activity was found at a dose of 500 mg/kg of crude extract (15.54 ± 0.37 sec) which differed significantly (P <0.01 and P <0.001) with that of the standard drug ketorolac (16.38 ± 0.27 sec). In acetic acid induced writhing test, the crude methanolic extract showed significant (P <0.01 and P <0.001) analgesic potential at doses 250 and 500 mg/kg body weight (45.98 and 56.32% inhibition, respectively), where ketorolac showed 64.36% inhibition. In anti-inflammatory activity test, the crude methanolic extract showed significant (P <0.001) potential at doses 250 and 500 mg/kg body weight (1.51 ± 0.04 and 1.47 ± 0.03 mm paw edema, respectively), where ketorolac showed 1.64 ± 0.05 mm edema after 3 h of carrageenan injection. In antipyretic activity assay, the crude extract showed notable reduction in body temperature (32.78 ± 0.46°C) at dose of 500 mg/kg-body weight, when the standard (at dose 150 mg/kg-body weight) exerted 33.32 ± 0.67°C temperature after 3 h of administration. CONCLUSIONS: Our results yield that the crude hydroalcoholic extract has better effects than the other in all trials. In the context, it can be said that the leaves of L. glutinosa possess remarkable pharmacological effects, and justify its traditional use as analgesic, antipyretic, anti-inflammatory, and thrombolytic agent.

Antidepressant activity of Litsea glaucescens essential oil: identification of ß-pinene and linalool as active principles.

ETHNOPHARMACOLOGICAL RELEVANCE: Litsea glaucescens (Lauraceae) has been used in Mexican Traditional Medicine to relieve illness related to central nervous system, such as epilepsy, fright and sadness. In this study, L. glaucescens essential oil properties on central nervous system were evaluated in mice using different behavioral tests. MATERIALS AND METHODS: The essential oil was obtained by hydrodistillation and analyzed by GC/MS. Identification of major compounds was also carried out by comparison with authentic samples. The psychopharmacological profile of L. glaucescens essential oil, and some its major compounds, were evaluated in mice using several experimental models: forced swimming test (FST: Antidepressant-like activity), open field test (OFT: Spontaneous locomotor activity), elevated plus-maze (EPM: Anxiolytic-like activity), exploratory cylinder (ECT: Sedative-like activity), rotarod (motor coordination) and traction performance (myo-relaxant effect) the essential oil and active principles was administered intraperitoneally. RESULTS: The essential oil showed antidepressant-like activity at doses of 100 and 300 mg/Kg. The monoterpenes ß-pinene and linalool were identified as the two main active principles of the essential oil, and showed antidepressant-like and sedative-like activity. Eucalyptol, limonene and α-pinene they did not show antidepressant-like activity, and were not further tested. CONCLUSIONS: L. glaucescens essential oil showed antidepressant activity, ß-pinene and linalool were identified as its active principles. These results support the use of L. glaucescens in Mexican Traditional Medicine for the treatment of sadness.

Actividad biológica y composición química de especies del género Litsea en Mesoamérica: Una revisión / Biological activity and chemical composition of Litsea´s genus species in Mesoamerica: a review

El presente trabajo es una revisión sobre la actividad biológica de extractos naturales con potencial medicinal o agroindustrial, en el cual se muestra que las especies vegetales del género Litsea distribuidas en Mesoamérica podrían constituir una fuente importante en el desarrollo de potenciales medicamentos y como biocontroladores tales como antitumoral, antiviral, antimicrobiana, antiinflamatoria, antioxidante, y en control fitosanitario como insecticida, plaguicida y repelente., entre otras. El desarrollo de un producto natural involucra la selección de la especie empleando criterios etnobotánicos o por bioprospección, los métodos de extracción estudio químico para el aislamiento y elucidación estructural de las moléculas responsables de la actividad y pruebas biológicas realizadas mediante ensayos in vitro, in vivo, toxicológicos y clínicos...