RESUMEN
The present study thus aimed at the development and physicochemical characterization of solid lipid nanoparticles loaded with crude extract of Piper corcovadensis roots (SLN - CEPc) and chitosan - coated solid lipid nanoparticles loaded with crude extract of P. corcovadensis roots (C - SLN - CEPc), as well as the determination of its antimycobacterial activity against Mycobacterium tuberculosis H37Rv, its cytotoxicity against the Vero cell line and evaluation in the hemolysis assay. Both formulat ions containing the encapsulated extract showed high encapsulation efficiency, formed by a monodispersed system with small and spherical particles, and there was no aggregation of particles. In the biological assays, SLN - CEPc and C - SLN - CEPc showed promisin g anti - M. tuberculosis activity with a minimum inhibitory concentration (MIC) of 12.5 µg/mL, whereas the cytotoxic concentrations obtained at 50% (CC 50 ) in Vero cells were 60.0 and 70.0 µg/mL, respectively. Therefore, nanoencapsulation showed satisfactory results, justifying its usage in the development of new products.
El presente estudio apuntó al desarrollo y caracterización fisicoquímica de na nopartículas lípidas en estado sólido, cargadas con extracto crudo de raíz de Piper c orcovadensis (SLN - CEPc) y nanopartículas lípidas en estado sólido cubiertas con quitosano cargadas co n extracto crudo de raíz de P. corcovadensis (C - SLN - CEPc), así como la determinación de su actividad antimico bacterial contra Mycobacterium tuberculosis H37Rv, su citotoxicidad contra la línea celular Vero y su evaluación en ensayo de hemólisis. Ambas formulaciones que contenían el extracto encapsulado mostraron alta eficien cia de encapsulación, formado por un sistema monodispersado con pequeñas partículas esféricas, y no hubo agregación de partículas. En los ensayos biológicos, SLN - CEPc y C - SLN - CEPc mostraron un a prometedora actividad anti - M. tuberculosis con una mínima conc entración inhibitoria (MIC) de 12,5 µg/mL, mientras que las concentraciones citotóxicas obtenidas al 50% (CC 50 ) en células Vero estuvo en 60,0 y 70,0 µg/mL, respectivamente. Por lo tanto, la nanoencapsulación mostró resultados satisfactorios, justificando su uso en el desarrollo de nuevos productos.
Asunto(s)
Extractos Vegetales/administración & dosificación , Sistemas de Liberación de Medicamentos , Piper/química , Antibacterianos/administración & dosificación , Mycobacterium tuberculosis/efectos de los fármacos , Temperatura , Portadores de Fármacos , Cromatografía Líquida de Alta Presión , Raíces de Plantas , Quitosano , Nanopartículas , LípidosRESUMEN
Abstract The therapeutic drugs to treat Herpes simplex virus (HSV) infections have toxic side effects and there has been an emergence of drug-resistant strains. Therefore, the search for new treatments for HSV infections is mounting. In the present study, semi-solid formulations containing a crude hydroethanolic extract (CHE) from Schinus terebinthifolia were developed. Skin irritation, cutaneous permeation, and in vivo therapeutic efficacy of the formulations were investigated. Treatment with the ointment formulations did not result in any signs of skin irritation while the emulsions increased the thickness of the epidermis in Swiss mice. The cutaneous permeation test indicated that the CHE incorporated in the formulations permeated through the skin layers and was present in the epidermis and dermis even 3 h after topical application. In vivo antiviral activity in BALB/c mice treated with the CHE ointments was better than those treated with the CHE emulsions and did not significantly differ from an acyclovir-treated group. Taken together, this suggests that the incorporation of CHE in the ointment may be a potential candidate for the alternative topical treatment of herpetic lesions.
Asunto(s)
Preparaciones Farmacéuticas/análisis , Simplexvirus/clasificación , Herpesvirus Humano 1/clasificación , Anacardiaceae/efectos adversos , Antivirales/efectos adversos , Aciclovir/antagonistas & inhibidores , Eficacia , Emulsiones/efectos adversosRESUMEN
Phytochemical investigation of Chromolaena laevigata led to the isolation of a new cadinene-sesquiterpene, chromolaevigone glucoside (1), along with nine known compounds: daucosterol (2), stigmasterol glycoside (3), stigmasterol (4), ß-sitosterol (5), pilloin (6), gonzalitosin I (7), quercetin-3-O-α-rhamnopyranoside (8), 7,7-dihydroxy-calamen-12-oic acid lactone (9) and trachelanthic acid (10). Others 11 known compounds were identified by UHPLC-HRMS/MS. These compounds are being described for the first time in this species, with the exception of cadinene 9. Furthermore, due to the limitation of pharmacological studies, antiproliferative, antiviral, and antimicrobial activities of C. laevigata were evaluated. The best results in the cytotoxicity, antimicrobial and antiproliferative tests, presenting GI50 values on ovarian tumour cells (OVCAR-03) of 1.9 µg mL-1 and kidney (786-0) of 2.5 µg mL-1 were observed for the hexanic fraction.[Figure: see text].
Asunto(s)
Asteraceae , Chromolaena , Sesquiterpenos , Fitoquímicos/farmacología , Componentes Aéreos de las PlantasRESUMEN
There is an increasing demand for fungi control in grains, especially toxigenic. Also, there is growing concern on the use of synthetic fungicides; thus alternatives are needed. The aim of this study was to evaluate the antifungal and antimycotoxigenic action of essential oils (EOs) from Zingiber officinale, Cinnamomum zeylanicum and Cymbopogon martinii against Fusarium verticillioides, a spoilage and toxigenic fungus. Essential oils were first chemically characterised by gas chromatography coupled to mass spectrometry, and their antioxidant potential was measured by the DPPH, ABTS and FRAP methods. Minimum inhibitory concentration (MIC) and disc diffusion were used to assess antifungal activity. Scanning electron microscopy was used to evaluate morphological changes in the fungus. Antimycotoxigenic activity of the EOs against the production of fumonisin B1 and B2 by F. verticillioides was evaluated using ultra-high-performance liquid chromatography system. Z. officinale, C. zeylanicum and C. martinii EOs were predominantly composed by zingiberene and geranial; eugenol; and geraniol, respectively. All the EOs had high antioxidant power, especially that from C. zeylanicum. The MICs were 250, 500 and 2,000 µg mL-1 for C. zeylanicum, C. martinii and Z. officinale EOs, respectively. Mycelial reduction of F. verticillioides was observed when EOs were used, and the lowest activity was detected in the Z. officinale EO. Overall, the tested EOs promoted structural damage to the fungal cell wall, decreased conidia size and mycelial reduction. Antimycotoxigenic evaluation of the EOs evidenced a significant reduction (p < .05) in the production of fumonisins B1 and B2 with all the EOs evaluated in the study. These results suggest that especially C. zeylanicum and C. martinii EOs are highly useful for controlling F. verticillioides and fumonisins production.
Asunto(s)
Antifúngicos/farmacología , Fumonisinas/antagonistas & inhibidores , Fusarium/efectos de los fármacos , Aceites Volátiles/farmacología , Extractos Vegetales/farmacología , Antifúngicos/química , Antifúngicos/aislamiento & purificación , Cinnamomum zeylanicum/química , Cymbopogon/química , Zingiber officinale/química , Pruebas de Sensibilidad Microbiana , Estructura Molecular , Aceites Volátiles/química , Aceites Volátiles/aislamiento & purificación , Extractos Vegetales/química , Extractos Vegetales/aislamiento & purificaciónRESUMEN
The antiviral potential of natural polysaccharide compounds has been demonstrated, especially against enveloped viruses and members of the Herpesviridae family. Two polysaccharide fractions obtained from Stevia rebaudiana (Bertoni) leaves, that were active against Herpes simplex virus type 1 (HSV-1) were studied to investigate their mode of action. Both polysaccharides - SFW (crude faction) and SSFK (homogeneous alkaline fraction) - exerted antiviral effects on the initial stages of HSV-1 infection by inhibiting viral adsorption and penetration. When added after virus internalization, both fractions decreased plaque size. The effect of the fractions was confirmed by investigating viral glycoprotein expression. Based on the mode of action of the polysaccharides demonstrated in the present work and on their selectivity index, the polysaccharides obtained from S. rebaudiana could be an alternative treatment of infections caused by HSV-1.
Asunto(s)
Antivirales/aislamiento & purificación , Herpesvirus Humano 1/efectos de los fármacos , Extractos Vegetales/uso terapéutico , Hojas de la Planta/química , Stevia/química , Antivirales/farmacología , Herpesviridae/efectos de los fármacos , Extractos Vegetales/química , Extractos Vegetales/farmacología , Polisacáridos/aislamiento & purificación , Polisacáridos/farmacología , Polisacáridos/uso terapéuticoRESUMEN
Protozoa of the genus Phytomonas are harmful parasites to several agricultural crops of economic importance. Due to their recognized biological activity, crude extracts of Piper aduncum, P. crassinervium, P. hispidum, and P. amalago leaves, were tested using the microdilution plate technique to assess the antiparasitic potential against Phytomonas serpens. Results showed that the ethanolic crude extract of P. crassinervium and P. amalago presented the best inhibitory concentration for 50% of the cells (IC50), 16.5 µg mL-1 in chloroform phase, and 18 µg mL-1 in aqueous phase, respectively, after 48 h treatment. Cytotoxicity analyses were performed using the colorimetric method of sulforhodamine-B in LLCMK2 mammalian cells. The chloroform phase of P. crassinervium was subjected to the fractionation process, in which the ethyl acetate and dichloromethane fractions obtained better IC50 values. Scanning electron microscopy (SEM) images showed alterations in the cell membrane of the treated parasites. The data obtained indicate a potential antiparasitic effect of the Piper species analyzed against P. serpens, being considered promising candidates for formulations of bioproducts to control the parasite.(AU)
Protozoários do gênero Phytomonas são parasitas prejudiciais a várias culturas agrícolas de importância econômica. Devido a sua atividade biológica reconhecida, extratos brutos de folhas de Piper aduncum, P. crassinervium, P. hispidum e P. amalago, foram testadas pela técnica de microdiluição em placa para avaliar o seu potencial antiparasitário contra Phytomonas serpens. Os resultados mostraram que o extrato bruto etanólico de P. crassinervium e P. amalago apresentaram as melhores concentrações inibitórias para 50% das células (IC50), 16,5 µg mL-1 na fase clorofórmio e 18 µg mL-1 na fase aquosa, respectivamente, após 48 h de tratamento. Análises de citotoxicidade foram realizadas através do método colorimétrico da sulforodamina-B, em células de mamíferos LLCMK2. A fase clorofórmio de P. crassinervium foi submetida ao processo de fracionamento, no qual as frações acetato de etila e diclorometano obtiveram melhores valores de IC50. Imagens de microscopia eletrônica de varredura (MEV) mostraram alterações na membrana celular dos parasitas tratados com fase aquosa de P. amalago. Os dados obtidos indicam potencial efeito antiparasitário das espécies de Piper analisadas contra P. serpens, sendo consideradas candidatas promissoras para formulações de bioprodutos para controle do parasito.(AU)
Asunto(s)
Piperaceae , Antiparasitarios , Plantas Medicinales , Trypanosomatina/patogenicidad , Enfermedades de las Plantas/prevención & controlRESUMEN
ABSTRACT: Protozoa of the genus Phytomonas are harmful parasites to several agricultural crops of economic importance. Due to their recognized biological activity, crude extracts of Piper aduncum, P. crassinervium, P. hispidum, and P. amalago leaves, were tested using the microdilution plate technique to assess the antiparasitic potential against Phytomonas serpens. Results showed that the ethanolic crude extract of P. crassinervium and P. amalago presented the best inhibitory concentration for 50% of the cells (IC50), 16.5 µg mL-1 in chloroform phase, and 18 µg mL-1 in aqueous phase, respectively, after 48 h treatment. Cytotoxicity analyses were performed using the colorimetric method of sulforhodamine-B in LLCMK2 mammalian cells. The chloroform phase of P. crassinervium was subjected to the fractionation process, in which the ethyl acetate and dichloromethane fractions obtained better IC50 values. Scanning electron microscopy (SEM) images showed alterations in the cell membrane of the treated parasites. The data obtained indicate a potential antiparasitic effect of the Piper species analyzed against P. serpens, being considered promising candidates for formulations of bioproducts to control the parasite.
RESUMO: Protozoários do gênero Phytomonas são parasitas prejudiciais a várias culturas agrícolas de importância econômica. Devido a sua atividade biológica reconhecida, extratos brutos de folhas de Piper aduncum, P. crassinervium, P. hispidum e P. amalago, foram testadas pela técnica de microdiluição em placa para avaliar o seu potencial antiparasitário contra Phytomonas serpens. Os resultados mostraram que o extrato bruto etanólico de P. crassinervium e P. amalago apresentaram as melhores concentrações inibitórias para 50% das células (IC50), 16,5 µg mL-1 na fase clorofórmio e 18 µg mL-1 na fase aquosa, respectivamente, após 48 h de tratamento. Análises de citotoxicidade foram realizadas através do método colorimétrico da sulforodamina-B, em células de mamíferos LLCMK2. A fase clorofórmio de P. crassinervium foi submetida ao processo de fracionamento, no qual as frações acetato de etila e diclorometano obtiveram melhores valores de IC50. Imagens de microscopia eletrônica de varredura (MEV) mostraram alterações na membrana celular dos parasitas tratados com fase aquosa de P. amalago. Os dados obtidos indicam potencial efeito antiparasitário das espécies de Piper analisadas contra P. serpens, sendo consideradas candidatas promissoras para formulações de bioprodutos para controle do parasito.
RESUMEN
Daidzein (DZ) is a polyphenolic compound belonging to Biopharmaceutical Classification System class IV, which shows that it may have limited therapeutic effects due to its low solubility and poor bioavailability. This study aimed to obtain high-purity DZ and prepare and characterize different types of solid dispersions (SDs) in order to enhance aqueous solubility and bioavailability. Excipients were investigated in order to manufacture different types of solid dispersions (SDs). Second-generation solid dispersions (SG), third-generation solid dispersions (TG), and second- and third-generation pH-modulated solid dispersions (SD and TG pHM-SD) were produced via spray drying. The SDs were characterized and tested for in vitro DZ release and oral bioavailability. SDs have shown increased aqueous solubility and in vitro release rate. Solid-state characterization showed that DZ was in an amorphous state in most of the formulations. The enhanced aqueous solubility of TG-pHM SD was reflected by an increase in oral bioavailability, which significantly increased the maximum plasma concentration approximately 20-fold and decreased the time to reach the maximum plasma concentration. The production of pHM SDs that contain DZ via spray drying is a simple and effective approach for oral drug delivery, which has the potential to greatly reduce the dose and enhance therapeutics effects.
RESUMEN
Visceral leishmaniasis, caused by Leishmania infantum, is a neglected tropical disease, to which efforts in the innovation of effective and affordable treatments remain limited, despite the rising incidence in several regions of the world. In this work, the antileishmanial effects of sugiol were investigated in vitro. This compound was isolated from the bark of Cupressus lusitanica and showed promising activity against L. infantum. In spite of the positive results, it is known that the compound is a poorly water-soluble diterpene molecule, which hinders further investigation, especially in preclinical animal studies. Thus, in an alternative delivery method, sugiol was entrapped in glucan-rich particles obtained from Saccharomyces cerevisiae yeast cell walls (YCWPs). To evaluate the activity of sugiol, the experiments were divided into two parts: (i) the in vitro investigation of antileishmanial activity of free sugiol against L. infantum promastigotes after 24, 48, and 72 h of treatment and (ii) the evaluation of antileishmanial activity of sugiol entrapped in glucan-rich particles against intracellular L. infantum amastigotes. Free sugiol induced the cell-death process in promastigotes, which was triggered by enhancing cytosolic calcium level and promoting the autophagy up to the first 24 h. Over time, the presence of autophagic vacuoles became rarer, especially after treatment with lower concentrations of sugiol, but other cellular events intensified, like ROS production, cell shrinkage, and phosphatidylserine exposure. Hyperpolarization of mitochondrial membrane potential was found at 72 h, induced by the mitochondria calcium uptake, causing an increase in ROS production and lipid peroxidation as a consequence. These events resulted in the cell death of promastigotes by secondary necrosis. Sugiol entrapped in glucan-rich particles was specifically recognized by dectin-1 receptor on the plasma membrane of macrophages, the main host cell of Leishmania spp. Electron micrographs revealed particles containing sugiol within the infected macrophages and these particles were active against the intracellular L. infantum amastigotes without affecting the host cell. Therefore, the YCWPs act like a Trojan horse to successfully deliver sugiol into the macrophage, presenting an interesting strategy to deliver water-insoluble drugs to parasitized cells.
Asunto(s)
Antiprotozoarios/farmacología , Muerte Celular/efectos de los fármacos , Diterpenos/farmacología , Leishmania infantum/efectos de los fármacos , Leishmaniasis Visceral/tratamiento farmacológico , Animales , Autofagia/efectos de los fármacos , Calcio/metabolismo , Pared Celular , Modelos Animales de Enfermedad , Femenino , Glucanos , Lectinas Tipo C , Leishmania infantum/citología , Leishmania infantum/patogenicidad , Macrófagos/metabolismo , Potencial de la Membrana Mitocondrial , Ratones Endogámicos BALB C , Mitocondrias/metabolismo , Membranas Mitocondriales/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Saccharomyces cerevisiaeRESUMEN
BACKGROUND: Herpes simplex type 1 (HSV-1) is widely distributed throughout the world's population. The virus spreads through direct contact with an infected individual. After primary infection, the virus remains in a latent state, and the recurrence of herpetic lesions is common. Standard treatment is performed with nucleoside analogues, but the selection of resistant strains have occurred, thus requiring the continual search for new antiviral agents. Plant extracts, fractions, and isolated compounds are a good source for studying possible antiviral compounds. HYPOTHESIS: Among plants with antiviral activity, the crude extract of aerial parts of Tanacetum parthenium (L.) Sch.Bip. (Asteraceae) have previously shown to inhibit HSV-1 infection in vitro. METHODS: The present study investigated the chemical composition of a crude hydroethanolic extract (CHE) of T. parthenium, and in vivo safety and therapeutic efficacy against HSV-1 infection. RESULTS: Liquid chromatography-mass spectrometry showed that the CHE was composed of phenolic acids (chlorogenic acids) and sesquiterpene lactones (parthenolide). Acute and subchronic toxicity and genotoxicity tests in vivo showed that oral CHE administration did not result in signs of toxicity, with no genotoxic potential. The CHE was also safe for topical administration, in which no irritation of the epidermis was observed in treated animals. Tests of topical and oral therapeutic efficacy showed that the CHE was effective against HSV-1 infection. Topical administration was the most effective, the results for which were comparable to acyclovir. CONCLUSION: These findings indicate that the CHE from aerial parts of Tanacetum parthenium has in vivo anti-HSV-1 activity and is safe for oral and topical application.
Asunto(s)
Antivirales/toxicidad , Antivirales/uso terapéutico , Herpes Simple/tratamiento farmacológico , Herpesvirus Humano 1/efectos de los fármacos , Extractos Vegetales/toxicidad , Tanacetum parthenium/química , Tanacetum parthenium/toxicidad , Animales , Antivirales/farmacología , Ratones , Modelos Animales , Extractos Vegetales/química , Espectrometría de Masas en TándemRESUMEN
BACKGROUND/AIMS: Parthenolide is a sesquiterpene lactone that is present in plants of the Tanacetum genus, for which many biological effects have already been reported, including antiherpetic activity. Although the effectiveness of parthenolide against Herpes simplex virus 1 (HSV-1) has already been demonstrated, such findings are still controversial. The objective of this study was to investigate the ways in which parthenolide exerts anti-HSV-1 activity. METHODS: The cytotoxicity and antiviral activity of parthenolide were determined by the MTT method and plaque reduction assay, respectively. The expression of cell and viral proteins during the treatment of infected cells was investigated by Western blot. RESULTS: Both strains of HSV-1 were sensitive to parthenolide, and parthenolide was active only after penetration of the virus into the host cell. The expression of p65 protein decreased, the expression of caspases 8 and 9 increased, and the expression of c-Jun N-terminal kinase (JNK) and p38 protein was altered in infected cells after parthenolide treatment, resulting in lower cell survival. The low expression of viral proteins gB, gD, and ICP0 confirmed the reduction of HSV-1 particle production. CONCLUSION: Parthenolide exerts anti-HSV-1 activity by impairing cell viability, which consequently interferes with the efficient infection and production of new viral particles.
Asunto(s)
Antivirales/farmacología , Herpes Simple/tratamiento farmacológico , Herpesvirus Humano 1/efectos de los fármacos , Sesquiterpenos/farmacología , Tanacetum/química , Animales , Caspasas/efectos de los fármacos , Caspasas/metabolismo , Supervivencia Celular/efectos de los fármacos , Chlorocebus aethiops , Herpes Simple/virología , Herpesvirus Humano 1/fisiología , Humanos , FN-kappa B/efectos de los fármacos , FN-kappa B/metabolismo , Componentes Aéreos de las Plantas/química , Células Vero , Proteínas Virales/efectos de los fármacos , Proteínas Virales/metabolismo , Replicación Viral/efectos de los fármacosRESUMEN
Herpes simplex virus infections persist throughout the lifetime of the host and affect more than 80â% of the humans worldwide. The intensive use of available therapeutic drugs has led to undesirable effects, such as drug-resistant strains, prompting the search for new antiherpetic agents. Although diverse bioactivities have been identified in Schinus terebinthifolia, its antiviral activity has not attracted much attention. The present study evaluated the antiherpetic effects of a crude hydroethanolic extract from the stem bark of S. terebinthifolia against Herpes simplex virus type 1 in vitro and in vivo as well as its genotoxicity in bone marrow in mammals and established the chemical composition of the crude hydroethanolic extract based on liquid chromatography-diode array detector-mass spectrometry and MS/MS. The crude hydroethanolic extract inhibited all of the tested Herpes simplex virus type 1 strains in vitro and was effective in the attachment and penetration stages, and showed virucidal activity, which was confirmed by transmission electron microscopy. The micronucleus test showed that the crude hydroethanolic extract had no genotoxic effect at the concentrations tested. The crude hydroethanolic extract afforded protection against lesions that were caused by Herpes simplex virus type 1 in vivo. Liquid chromatography-diode array detector-mass spectrometry and MS/MS identified 25 substances, which are condensed tannins mainly produced by a B-type linkage and prodelphinidin and procyanidin units.
Asunto(s)
Anacardiaceae/química , Antivirales/farmacocinética , Herpes Simple/tratamiento farmacológico , Herpesvirus Humano 1/efectos de los fármacos , Extractos Vegetales/farmacología , Animales , Antivirales/química , Antivirales/aislamiento & purificación , Antivirales/farmacología , Supervivencia Celular/efectos de los fármacos , Chlorocebus aethiops , Cromatografía Liquida , Femenino , Herpes Simple/virología , Herpesvirus Humano 1/ultraestructura , Humanos , Masculino , Ratones , Ratones Endogámicos BALB C , Microscopía Electrónica de Transmisión , Corteza de la Planta/química , Extractos Vegetales/química , Extractos Vegetales/aislamiento & purificación , Espectrometría de Masas en Tándem , Taninos/análisis , Taninos/química , Células VeroRESUMEN
Leishmaniasis is a neglected infection that is caused by Leishmania protozoa, affecting millions of people worldwide, mainly in tropical and subtropical regions. This disease has different clinical forms: cutaneous, mucocutaneous, and visceral. The drugs that are currently available for the treatment of this infection have limitations, such as high toxicity, long-term treatment, and leads to drug-resistant strains. Numerous studies, in various experimental models, have sought to develop more effective and less toxic chemotherapeutic agents against leishmaniasis. In the present study, we evaluated the mechanism of cell death that is induced by n-benzyl 1-(4-methoxy)phenyl-9H-ß-carboline-3-carboxamide (C5) against Leishmania amazonensis. C5 increased reactive oxygen species production, depolarization of the mitochondrial membrane, DNA fragmentation, decrease of cell volume, lipoperoxidation, the accumulation of lipid bodies, and acidic vesicular organelles (AVOs) and caused the intense formation of autophagic compartments in L. amazonensis promastigotes. The results indicate that C5 causes L. amazonensis death through different pathways.
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Antiprotozoarios/farmacología , Carbolinas/farmacología , Leishmania mexicana/efectos de los fármacos , Leishmaniasis Cutánea/tratamiento farmacológico , Antiprotozoarios/química , Carbolinas/química , Fragmentación del ADN/efectos de los fármacos , Humanos , Leishmania mexicana/citología , Leishmania mexicana/genética , Leishmania mexicana/metabolismo , Leishmaniasis Cutánea/parasitología , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Estrés Oxidativo/efectos de los fármacos , Especies Reactivas de Oxígeno/metabolismoRESUMEN
The aim of the present work was to develop a topical delivery system that contains Brazilian green propolis extract (PE-8) to increase efficiency and convenience when applied to herpetic lesions. The cytotoxicity and antiherpetic activity was determined in vitro and in vivo. The PE-8 was added to a system that contained poloxamer 407 and carbopol 934P. The in vitro characterization of the system included rheological studies, texture profile analysis, and mucoadhesion analysis. The PE-8 inhibited the virus during the phase of viral infection, induced virion damage, and exhibited an ability to protect cells from viral infection. The system had advantageous mucoadhesive properties, including a suitable gelation temperature of approximately 25°C for topical delivery, a desirable textural profile, and pseudoplastic behavior. The in vitro release study showed a rapid initial release of the PE-8 in the first 3 h, and the rate of drug release remained constant for up to 24 h. The system appeared to be macroscopically and microscopically innocuous to skin tissue. Therefore, the mucoadhesive thermoresponsive system that contained the PE-8 appears to be promising for increasing bioavailability and achieving prolonged release of the PE-8 when applied to skin lesions caused by herpes simplex virus type 1.
Asunto(s)
Antivirales/administración & dosificación , Portadores de Fármacos/química , Herpes Simple/tratamiento farmacológico , Herpesvirus Humano 1/efectos de los fármacos , Própolis/administración & dosificación , Acrilatos/química , Adhesividad , Animales , Antivirales/química , Antivirales/uso terapéutico , Antivirales/toxicidad , Supervivencia Celular/efectos de los fármacos , Chlorocebus aethiops , Relación Dosis-Respuesta a Droga , Liberación de Fármacos , Femenino , Herpes Simple/metabolismo , Masculino , Ratones , Ratones Endogámicos BALB C , Mucosa Bucal/efectos de los fármacos , Mucosa Bucal/metabolismo , Mucosa Bucal/virología , Poloxámero/química , Própolis/química , Própolis/uso terapéutico , Própolis/toxicidad , Reología , Piel/efectos de los fármacos , Piel/metabolismo , Piel/virología , Temperatura , Células VeroRESUMEN
The ethnomedicinal plant Curatella americana L. (Dilleniaceae) is a common shrub in the Brazilian cerrado, in which crude extract showed antifungal activity in a preliminary study. In this work, the antifungal and cytotoxic properties of the crude extract, fractions, and isolated compounds from C. americana were evaluated against the standard yeast strains Candida albicans, C. tropicalis, and C. parapsilosis, clinical isolates, and fluconazole-resistant strains. The combinatory effects between subfractions and isolated compounds and effects on cell morphology, virulence factors, and exogenous ergosterol were also evaluated. The MIC obtained against the Candida species including fluconazole-resistant strain ranged from 15.3 to 31.3 µg/mL for crude extract, 3.9 to 15.6 µg/mL for ethyl acetate fraction, and 7.8 to 31.3 µg/mL for subfractions. The isolated compounds identified as 4'-O-methyl-catechin, epicatechin-3-O-gallate, and 4'-O-methyl-catechin-3-O-gallate showed lower antifungal activity than the crude extract and fractions (MIC ranging from 31.3 to 125.0 µg/mL). The addition of exogenous ergosterol to yeast culture did not interfere in the antifungal activity of the extract and its fractions. Synergistic antifungal activity was observed between subfractions and isolated compounds. The effects on virulence factors and the different mechanisms of action compared to fluconazole and nystatin suggest that this ethnomedicinal plant may be an effective alternative treatment for candidiasis.
RESUMEN
Trypanosoma cruzi is the causative agent of Chagas' disease, a parasitic disease that remains a serious health concern with unsatisfactory treatment. Drugs that are currently used to treat Chagas' disease are partially effective in the acute phase but ineffective in the chronic phase of the disease. The aim of the present study was to evaluate the antitrypanosomal activity and morphological, ultrastructural and biochemical alterations induced by a new molecule, 4-nitrobenzaldehyde thiosemicarbazone (BZTS), derived from S-(-)-limonene against epimastigote, trypomastigote and intracellular amastigote forms of T. cruzi. BZTS inhibited the growth of epimastigotes (IC50 = 9·2 µ m), intracellular amastigotes (IC50 = 3·23 µ m) and inhibited the viability of trypomastigotes (EC50 = 1·43 µ m). BZTS had a CC50 of 37·45 µ m in LLCMK2 cells. BZTS induced rounding and distortion of the cell body and severely damaged parasite mitochondria, reflected by extensive swelling and disorganization in the inner mitochondrial membrane and the presence of concentric membrane structures inside the organelle. Cytoplasmic vacuolization, endoplasmic reticulum that surrounded organelles, the loss of mitochondrial membrane potential, and increased mitochondrial O2 â¢- production were also observed. Our results suggest that BZTS alters the ultrastructure and physiology of mitochondria, which could be closely related to parasite death.
Asunto(s)
Ciclohexenos/química , Estadios del Ciclo de Vida/efectos de los fármacos , Mitocondrias/efectos de los fármacos , Terpenos/química , Tripanocidas/farmacología , Trypanosoma cruzi/efectos de los fármacos , Animales , Benzaldehídos/química , Benzaldehídos/farmacología , Línea Celular , Retículo Endoplásmico/efectos de los fármacos , Retículo Endoplásmico/patología , Retículo Endoplásmico/ultraestructura , Células Epiteliales/efectos de los fármacos , Células Epiteliales/parasitología , Estadios del Ciclo de Vida/fisiología , Limoneno , Macaca mulatta , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Mitocondrias/metabolismo , Mitocondrias/patología , Mitocondrias/ultraestructura , Superóxidos/agonistas , Superóxidos/metabolismo , Tiosemicarbazonas/química , Tiosemicarbazonas/farmacología , Tripanocidas/química , Trypanosoma cruzi/crecimiento & desarrollo , Trypanosoma cruzi/metabolismo , Trypanosoma cruzi/ultraestructuraRESUMEN
The most bioactive soy isoflavones (SI), daidzein (DAI) and genistein (GEN) have poor water solubility, which reduces their bioavailability and health benefits and limits their use in industry. The goal of this study was to develop and characterize a new gelatin matrix to microencapsulate DAI and GEN from soy extract (SE) by spray drying, in order to obtain solid dispersions to overcome solubility problems and to allow controlled release. The influences of 1:2 (MP2) and 1:3 (MP3) SE/polymer ratios on the solid state, yield, morphology, encapsulation efficiency, particle size distribution, release kinetics and cumulative release were evaluated. Analyses showed integral microparticles and high drug content. MP3 and MP2 yield were 43.6% and 55.9%, respectively, with similar mean size (p > 0.05), respectively. X-ray diffraction revealed the amorphous solid state of SE. In vitro release tests showed that dissolution was drastically increased. The results indicated that SE microencapsulation might offer a good system to control SI release, as an alternative to improve bioavailability and industrial applications.
RESUMEN
The effect of a meropenem-ciprofloxacin combination (MCC) on the susceptibility of multidrug-resistant (MDR) Pseudomonas aeruginosa (MRPA) clinical isolates was determined using checkerboard and time-kill curve techniques. Structural changes and differential gene expression that resulted from the synergistic action of the MCC against one of the P. aeruginosa isolates (1071-MRPA]) were evaluated using electron microscopy and representational difference analysis (RDA), respectively. The differentially expressed, SOS response-associated, and resistance-associated genes in 1071-MRPA exposed to meropenem, ciprofloxacin, and the MCC were monitored by quantitative PCR. The MCC was synergistic against 25% and 40.6% of MDR P. aeruginosa isolates as shown by the checkerboard and time-kill curves, respectively. The morphological and structural changes that resulted from the synergistic action of the MCC against 1071-MRPA were a summation of the effects observed with each antimicrobial alone. One exception included outer membrane vesicles, which were seen in a greater amount upon ciprofloxacin exposure but were significantly inhibited upon MCC exposure. Cell wall- and DNA repair-associated genes were differentially expressed in 1071-MRPA exposed to meropenem, ciprofloxacin, and the MCC. However, some of the RDA-detected, resistance-associated, and SOS response-associated genes were expressed at significantly lower levels in 1071-MRPA exposed to the MCC. The MCC may be an alternative for the treatment of MDR P. aeruginosa. The effect of this antimicrobial combination may be not only the result of a summation of the effects of meropenem and ciprofloxacin but also a result of differential action that likely inhibits protective mechanisms in the bacteria.
Asunto(s)
Antibacterianos/farmacología , Ciprofloxacina/farmacología , Genes Bacterianos/efectos de los fármacos , Pseudomonas aeruginosa/genética , Tienamicinas/farmacología , Brasil , Recuento de Colonia Microbiana , ADN Bacteriano/biosíntesis , ADN Bacteriano/genética , Combinación de Medicamentos , Farmacorresistencia Bacteriana Múltiple/genética , Regulación Bacteriana de la Expresión Génica/efectos de los fármacos , Humanos , Meropenem , Pruebas de Sensibilidad Microbiana , Reacción en Cadena de la Polimerasa , Infecciones por Pseudomonas/microbiología , Pseudomonas aeruginosa/efectos de los fármacos , Pseudomonas aeruginosa/ultraestructura , ARN Bacteriano/biosíntesis , ARN Bacteriano/genéticaRESUMEN
Stryphnodendron adstringens has a high tannin content and is used as an antiseptic and antimicrobial and in the treatment of leucorrhea, gonorrhea, wound healing, and gastritis. The present study evaluated the toxic effects of the heptamer prodelphinidin (F2) from the stem bark of S. adstringens in rodents. In the acute toxicity test, the mice that received oral doses exhibited reversible effects, with an LD50 of 3.015 mg · kg(-1). In the chronic toxicity test at 90 days, Wistar rats were treated with different doses of F2 (10, 100, and 200 mg · kg(-1)). In the biochemical, hematological, and histopathological examinations and open-field test, the different dose groups did not exhibit significant differences compared with controls. The present results indicate that F2 from the stem bark of S. adstringens caused no toxicity with acute and chronic oral treatment in rodents at the doses administered.
RESUMEN
Yeasts of the Candida genus can colonize epithelium and mucosa of the vertebrate organisms; howeverthese can cause infection in a broad range of body sites. Candida species also can be found in drinking water and they are considered as a potential indicator of water quality. In this study were evaluated three methods to identify yeasts isolated from blotted water (seminested PCR, culture on CHROMagar Candida medium, and Candifast identification system). For this propose, we used 27 isolates fromblotted water and compared with 22 clinical isolates from vaginal fluid. Seminested PCR has shown specificity and sensitivity for identification of the Candida species. Candida albicans and Candida parapsilosis were the prevalent species from vaginal fluid and blotted water, respectively. Culture onCHROMagar and Candifast system had low agreement with snPCR (40.9% and 45.5%, respectively) in the yeasts identification from vaginal fluid. On the other hand, CHROMagar Candida can be used in the presumptive identification of yeasts isolated from bottled water and it had agreements percentage of 81.5% with snPCR method.
Leveduras do gênero Candida podem colonizar epitélio e mucosa dos organismos vertebrados, entretanto, podem causar infecções em vários lugares do corpo. Espécies de Candida, também, podem ser encontradas em água e são consideradas um potencial indicador da qualidade de água. Neste trabalho, foram avaliados três métodos de identificação de leveduras isoladas de água engarrafada (seminestedPCR, cultura no meio CHROMagar Candida e sistema de identificação Candifast). Foram utilizados 27 isolados de água engarrafada e comparados com 22 isolados clínicos de fluido vaginal. Seminested PCR tem mostrado especificidade e sensibilidade para a identificação das espécies de Candida. Candida albicans e Candida parapsilosis foram as espécies prevalentes do fluido vaginal e da água engarrafada,respectivamente. Cultura em CHROMagar e o sistema Candifast tiveram baixa concordância com snPCR(40,9% e 45,5%, respectivamente) na identificação de leveduras de fluido vaginal. Em contrapartida, CHROMagar Candida pode ser usado em identificação presuntiva de leveduras de água engarrafada apresentando concordância de 81,5% com o método snPCR.