RESUMO
Abstract Hamelia patens, is a plant traditionally used to treat a variety of conditions among the Huastec people of Mexico. The objective of this study is to characterize the phenolic content and critically examine the antimicrobial activity of leaf extracts H. patens, obtained by maceration, Soxhlet and percolation, using ethanol as 70% solvent. Phenolic compounds are characterized by liquid chromatography, coupled to a High Resolution Mass Spectrometry, and the antimicrobial activity was studied from the inhibitory effect of each extract for Escherichia coli, Staphylococcus aureus, Salmonella typhi and S. paratyphi, and by the Minimum Bactericidal Concentration, the percentage of activity and the Index of Bacterial Susceptibility of each extract. The phenolic compound identified in different concentrations in the three extracts was epicatechin. The extracts obtained by the three methods had antimicrobial activity, however, there was no significant difference (p < 0.05) between the Minimum Bactericidal Concentration of the extracts obtained by maceration, percolation and Soxhlet. The results of this study contribute to the body of knowledge on the use of extracts in controlling microorganisms with natural antimicrobials.
Assuntos
Fenóis/isolamento & purificação , Fenóis/farmacocinética , Extratos Vegetais/isolamento & purificação , Extratos Vegetais/farmacocinética , Hamelia/química , Fracionamento Químico/métodos , Antibacterianos/isolamento & purificação , Antibacterianos/farmacologia , Fenóis/química , Staphylococcus aureus/efeitos dos fármacos , Extratos Vegetais/química , Testes de Sensibilidade Microbiana , Escherichia coli/efeitos dos fármacos , México , Antibacterianos/químicaRESUMO
ABSTRACT The aim of this study was to optimize the dextranase production by fungus Pochonia chlamydosporia (VC4) and evaluate its activity in dextran reduction in sugarcane juice. The effects, over the P. chlamydosporia dextranase production, of different components from the culture medium were analyzed by Plackett-Burman design and central composite design. The response surface was utilized to determine the levels that, among the variables that influence dextranase production, provide higher production of these enzymes. The enzymatic effect on the removal of dextran present in sugarcane juice was also evaluated. It was observed that only NaNO3 and pH showed significant effect (p<0.05) over dextranase production and was determined that the levels which provided higher enzyme production were, respectively, 5 g/L and 5.5. The dextranases produced by fungus P. chlamydosporia reduced by 75% the dextran content of the sugarcane juice once treated for 12 hours, when compared to the control treatment.
Assuntos
Modelos Estatísticos , Saccharum/metabolismo , Dextranase/biossíntese , Hypocreales/enzimologia , Temperatura , Dextranos/metabolismo , Meios de Cultura/metabolismo , Eletroforese em Gel de Poliacrilamida , Ativação Enzimática , Sucos de Frutas e Vegetais/análise , Fracionamento Químico/métodos , Concentração de Íons de Hidrogênio , NitratosRESUMO
Sperm contains a wealth of cell surface receptors and ion channels that are required for most of its basic functions such as motility and acrosome reaction. Conversely, animal venoms are enriched in bioactive compounds that primarily target those ion channels and cell surface receptors. We hypothesized, therefore, that animal venoms should be rich enough in sperm-modulating compounds for a drug discovery program. Our objective was to demonstrate this fact by using a sperm-based phenotypic screening to identify positive modulators from the venom of Walterinnesia aegyptia. Methods Herein, as proof of concept that venoms contain interesting compounds for sperm physiology, we fractionated Walterinnesia aegyptia snake venom by RP-HPLC and screened for bioactive fractions capable of accelerating mouse sperm motility (primary screening). Next, we purified each compound from the positive fraction by cation exchange and identified the bioactive peptide by secondary screening. The peptide sequence was established by Edman sequencing of the reduced/alkylated compound combined to LC-ESI-QTOF MS/MS analyses of reduced/alkylated fragment peptides following trypsin or V8 protease digestion. Results Using this two-step purification protocol combined to cell phenotypic screening, we identified a new toxin of 7329.38 Da (actiflagelin) that activates sperm motility in vitro from OF1 male mice. Actiflagelin is 63 amino acids in length and contains five disulfide bridges along the proposed pattern of disulfide connectivity C1-C5, C2-C3, C4- C6, C7-C8 and C9-C10. Modeling of its structure suggests that it belongs to the family of three finger toxins with a noticeable homology with bucandin, a peptide from Bungarus candidus venom. Conclusions This report demonstrates the feasibility of identifying profertility compounds that may be of therapeutic potential for infertility cases where motility is an issue.(AU)
Assuntos
Animais , Masculino , Ratos , Motilidade dos Espermatozoides , Espermatozoides/química , Venenos Elapídicos/isolamento & purificação , Venenos Elapídicos/uso terapêutico , Fosfolipases A2 , Acetilcolinesterase , Espectrometria de Massas em Tandem/métodos , Fracionamento Químico/métodos , CamundongosRESUMO
Los contaminantes del aire han sido y siguen siendo, los principales factores que contribuyen a las enfermedades crónicas como el asma y enfermedades cardiovasculares. La contaminación del aire por material particulado (PM) es un problema mundial y en los últimos años, el PM se ha convertido en un tema importante de investigación ya que tiene un impacto negativo significativo en la salud humana; el PM es generado por las actividades industriales y tubos de escape de vehículos de motor. Sin embargo, diversos componentes nocivos del PM, como los hidrocarburos aromáticos policiclicos (HAP) en general, son sospechosos de ser carcinogénicos. Este trabajo tiene como objetivo identificar los HAP presentes en el PM2.5 del aire de Cúcuta, extraídos por primera vez, mediante el sistema diclorometano-etanol-tolueno e investigar la importancia del fraccionamiento de la materia organica del PM2.5 para detectar los HAP presentes en las fracciones del PM2.5. La identificación de los HAP considerados como contaminantes prioritarios y reconocidos por su afectación a la salud de la población se realizó, mediante cromatografía de gases con detector FID. Los efectos genotoxicos de la materia orgánica del PM2.5 extraída con una mezcla de DCM-etanol-tolueno fueron evaluados mediante el ensayo Cometa.
Air pollutants have been and still are the main factors that contribute to chronic diseases such as asthma and cardiovascular disease. Air pollution by particulate matter (PM) is a global problem and in recent years, the PM has become an important research topic since it has a significant negative impact on human health; the PM is generated by industrial activities and exhaust pipes of motor vehicles. However, various harmful components of PM such as polycyclic aromatic hydrocarbons (PAHs) in general, are suspected of being carcinogenic. This work aims to identify the PAHs present in the PM 2.5 air Cúcuta, first extracted by the dichloromethane-ethanol-toluene system and investigate the importance of organic matter fractionation of PM 2.5 to detect PAHs present in the fractions of PM 2.5. The identification of PAHs considered as priority pollutants and recognized for their effects on health of the population was performed by gas chromatography with FID detector. The genotoxic effects of PM2.5 organic matter, extracted with a mixture of DCM-ethanol-toluene, was evaluated by the Comet assay.
Assuntos
Humanos , Carcinógenos Ambientais , Genotoxicidade , Hidrocarbonetos Policíclicos Aromáticos/toxicidade , Fracionamento Químico/métodos , Colômbia , Ensaio Cometa/métodos , Poluição AmbientalRESUMO
Kanihua (Chenopodium pallidicaule Aellen) is a Chenopodiacea of the andean region, that contains between 15 and 19% protein, with essential amino acids. The objective of the study was to fractionate and electrophoretically characterize the proteins of kanihua seed varieties Cupi-Sayhua and Ramis. In the whole meal, the proximal analysis and fractionation were performed, and the flour was fractionated by five techniques according to Osborne solubility to obtain albumins, globulins, prolamins andglutelins. The methodology, solvents and extraction time were optimized; and the electrophoretic profiles of the fractions were identified. The highest protein content (p≤ 0.05) was of kanihua flour and its protein fractions, compared to kiwicha and wheat. The highest percent yield (p≤ 0.05) during 1 h of sequential extraction of the protein fractions, was obtained with the Rodriguez y et.al., technique for albumins and glutelins, and with the technique described by Barba de la Rosa y et.al., for globulins and prolamins. The following results were foundin Ramis and Cupi-Sayhua kanihua: albumins: 15.4±0.3 and 15.8±0.3%, globulins 7S: 24.1±0.5 and 26.3±1.0%, globulins 11S: 25.7±1.0 and 26.7±1.0%, prolamins: 9.6±0.1 and 9.9±0.5% and glutelins: 22.9±0.1 and 21.5±1.4%, respectively The electrophoretic profile showedpatterns similar in number of bands and differences in concentration in both varieties.
La kañihua (Chenopodium pallidicaule Aellen) es una Chenopodiacea de la región andina, que contiene entre 15 y 19% de proteínas, con aminoácidos esenciales. El objetivo del presente estudio fue fraccionar y caracterizar electroforéticamente las proteínas de la semilla de kañihua variedades Ramis y Cupi-Sayhua. En harina integral se realizó el análisis proximal y fraccionamiento, luego, la harina se fraccionó mediante cinco técnicas según la solubilidad de Osborne para obtener albuminas, globulinas, prolaminas y glutelinas. Se optimizó la metodología, solventes y tiempo de extracción; e identificaron los perfiles electroforéticos de las fracciones. El mayor contenido proteínico (p≤ 0,05) fue de la harina de kañihua y sus fracciones proteicas, en comparación a kiwicha y trigo. El mayor rendimiento porcentual (p≤ 0,05) durante 1 h de extracción secuencial de las fracciones proteicas, se obtuvo con la técnica de Rodríguez y et.al., para albuminas y glutelinas, y con la técnica de Barba de la Rosa y et.al., para globulinas y prolaminas. Se encontró en kañihua Ramis y Cupi-Sayhua, albúminas: 15,4±0,3 y 15,8+0,3%; globulinas: 7S 24,1±0,5 y 26,3+1,0%; globulinas 11S: 25,7+1,0 y 26,7+1,0%; prolaminas: 9,6+0,1 y 9,9+0,5% y glutelinas: 22,9+0,1 y 21,5+1,4%, respectivamente. El perfil electroforético mostró patrones similares en número de bandas y diferentes en concentración en ambas variedades.
Assuntos
Proteínas , Grão Comestível , Eletroforese , Fracionamento Químico , Valor NutritivoRESUMO
Abstract A bioassay-guided fractionation of two samples of Brazilian red propolis (from Igarassu, PE, Brazil, hereinafter propolis 1 and 2) was conducted in order to determine the components responsible for its antimicrobial activity, especially against Candida spp. Samples of both the crude powdered resin and the crude ethanolic extract of propolis from both locations inhibited the growth of all 12 tested Candida strains, with a minimum inhibitory concentration of 256 µg/mL. The hexane, acetate and methanol fractions of propolis 1 also inhibited all strains with minimum inhibitory concentration values ranging from 128 to 512 µg/mL for the six bacteria tested and from 32 to 1024 µg/mL for the yeasts. Similarly, hexane and acetate fractions of propolis sample 2 inhibited all microorganisms tested, with minimum inhibitory concentration values of 512 µg/mL for bacteria and 32 µg/mL for yeasts. The extracts were analyzed by HPLC and their phenolic profile allowed us to identify and quantitate one phenolic acid and seven flavonoids in the crude ethanolic extract. Formononetin and pinocembrin were the major constituents amongst the identified compounds. Formononetin was detected in all extracts and fractions tested, except for the methanolic fraction of sample 2. The isolated isoflavone formononetin inhibited the growth of all the microorganisms tested, with a minimum inhibitory concentration of 200 µg/mL for the six bacteria strains tested and 25 µg/mL for the six yeasts. Formononetin also exhibited fungicidal activity against five of the six yeasts tested. Taken together our results demonstrate that the isoflavone formononetin is implicated in the reported antimicrobial activity of red propolis.
Assuntos
Anti-Infecciosos/farmacologia , Candida/efeitos dos fármacos , Isoflavonas/isolamento & purificação , Isoflavonas/farmacologia , Própole/química , Anti-Infecciosos/isolamento & purificação , Brasil , Bactérias/efeitos dos fármacos , Fracionamento Químico , Cromatografia Líquida de Alta Pressão , Testes de Sensibilidade MicrobianaRESUMO
The objective of this work was investigate the synergistic, additive and antagonistic effects of fruit mixtures on total antioxidant capacities and bioactive compounds in tropical fruit juices, and optimize its formulation by the response surface methodology based on the responses: total polyphenols (TP), total antioxidant capacity (TAC), ascorbic acid content and sensorial acceptance. Camu-camu, acerola and acai were the major factors that influenced the antioxidant potential of the juice; and the yellow mombin showed a positive effect on the acceptance of the tropical juice. It was observed an antagonistic effect between acerola and camu-camu for the TAC response. The optimum formulation obtained was 20% acerola, 10% camu-camu, 10% yellow mombin, 10% cashew apple and 10% acai, which was responsible for a response of 155.46 mg.100 g-1 of ascorbic acid, 103.01 mg of GAE.100 g-1 of TP, 10.27 μM Trolox g-1 of TAC and approximately 6.1 of acceptance.
El objetivo de este trabajo fue investigar los efectos sinérgicos, aditivos y antagónicos de mezclas de diferentes frutas tropicales en la capacidad antioxidante total (TAC) y compuestos bioactivos presentes en los jugos mixtos, y optimizar su formulación por la metodología de superficie de respuesta basado en las evaluaciones de: polifenoles totales (TP), capacidad antioxidante total (TAC), contenido de ácido ascórbico y la aceptación sensorial. Camu-camu, acerola y acai fueron las frutas que más influyeron en el potencial antioxidante del jugo mixto; y el jobo mostró un efecto positivo en la aceptación del jugo mixto tropical. Se observó un efecto antagónico entre acerola y camu-camu para la TAC. La formulación óptima obtenida contenía 20% acerola, 10% de camu-camu, 10% el jobo, 10% de manzana de marañón y 10% de acai, la cual ha proporcionado contenidos medio de 155,46 mg.100 g-1 de ácido ascórbico, 103,01 mg de GAE.100 g-1 de TP, 10,27 mM Trolox g-1 de TAC y aproximadamente 6.1 de aceptación sensorial.
Assuntos
Humanos , Sucos de Frutas e Vegetais/análise , Anacardiaceae/química , Anacardium/química , Ananas/química , Antioxidantes/análise , Ácido Ascórbico/análise , Fracionamento Químico/métodos , Interações Medicamentosas , Euterpe/química , Mangifera/química , Polifenóis/análise , Projetos de Pesquisa , PaladarRESUMO
The present study investigated six varieties of locally grown wheat (Lasani, Sehar, Miraj-08, Chakwal-50, Faisalabad-08 and Inqlab) procured from Punjab Seed Corporation, Lahore, Pakistan for their proximate contents. On the basis of protein content and ready availability, Faisalabad-08 (FD-08) was selected to be used for the assessment of comparative efficiency of various methods used for gluten extraction. Three methods, mechanical, chemical and microbiological were used for the extraction of gluten from FD-08. Each method was carried out under ambient conditions using a drying temperature of 55°C. Mechanical method utilized four different processes viz:- dough process, dough batter process, batter process and ethanol washing process using standard 150 mesh. The starch thus obtained was analyzed for its proximate contents. Dough batter process proved to be the most efficient mechanical method and was further investigated using 200 and 300 mesh. Gluten content was determined using sandwich ω-gliadin enzyme-linked immunosorbent assay (ELISA).The results of dough batter process using 200 mesh indicated a starch product with gluten content of 678 ppm. Chemical method indicated high gluten content of more than 5000 ppm and the microbiological method reduced the gluten content from 2500 ppm to 398 ppm. From the results it was observed that no gluten extraction method is viable to produce starch which can fulfill the criteria of a gluten free product (20ppm).
El presente roduc seis variedades de trigo cultivado localmente (Lasani, Sehar, Miraj-08, Chakwal-50, Faisalabad-08 e Inqlab) obtenidos por sus contenidos proximales en Punjab Seed Corporation, Lahore, Pakistán. Sobre la base del contenido de roduct y su fácil disponibilidad, Faisalabad-08 (Fd-08) fue seleccionado para ser utilizado para la evaluación de la eficacia comparadativa de los diferentes métodos utilizados para la extracción de gluten. Tres métodos fueron utilizados para la extracción de gluten; mecánico, químico y microbiológico realizados en condiciones ambientales utilizando una roduct e de secado de 55°C. El método mecánico roduct cuatro procesos diferentes, a saber: proceso de masa, proceso de rebozado de masa, proceso de pasta y proceso de lavado en etanol empleando malla estándar de 150 mesh. El almidón obtenido se analizó por sus contenidos proximales. El contenido de gluten se roduct usando roduct ω-gliadina, ensayo de inmunoabsorción ligado a enzimas (ELISA). El proceso de rebozado de masa fue el método roduct más eficiente y se investigó adicionalmente usando malla 200 y 300 mesh. El proceso de rebozado de masa usando malla 200 generó un almidón con contenido de gluten de 678 ppm. El método químico produjo un contenido de gluten de más de 5.000 ppm, y el método microbiológico redujo el contenido de gluten de 2500 ppm a 398 ppm. A partir de estos resultados se roduct que ningún de estos métodos de extracción de gluten fue es viable para roduct almidón que pueda cumplir los criterios de un roduct libre de gluten (20 ppm).
Assuntos
Glutens/isolamento & purificação , Triticum/química , Fracionamento Químico/métodos , Análise de Alimentos/métodos , Manipulação de Alimentos/métodosRESUMO
A dinâmica da matéria orgânica do solo (MOS) pode ser melhor compreendida após identificadas suas frações lábeis (> 53 µm) e estáveis (< 53 µm). Diante da importância crescente do uso de técnicas de fracionamento físico da MOS, torna-se necessário avaliar se a recuperação de carbono nessas frações é influenciada pelos métodos utilizados para sua determinação. O objetivo deste trabalho foi avaliar os teores de carbono, por três métodos, em diferentes frações da matéria orgânica, num Latossolo Vermelho-Amarelo distrófico sob consórcio milho-forrageiras e em sistema solteiro. Foram determinados os teores de carbono das frações: (i) particulada da matéria orgânica (COP) e naquela (ii) associada aos minerais da matéria orgânica (COM). Determinou-se ainda, o teor de (iii) carbono orgânico total do solo (COT) na matéria orgânica não fracionada. Três métodos de determinação do carbono orgânico do solo foram utilizados: (i) Walkley & Black modificado, (ii) Mebius modificado, e (iii) combustão via seca, em analisador elementar (CHN). Foram utilizadas amostras de solos de três sistemas de manejo dispostos em delineamento em blocos casualizados, com três repetições para determinação do carbono orgânico do solo: (i) milho solteiro; (ii) milho consorciado com capim colonião (Panicum maximum cv. Aruana); e (iii) milho consorciado com braquiária (Brachiaria humidicola). As amostras de solo foram coletadas na profundidade de 0 5 cm. O método Walkley & Black apresentou menores valores de COT em relação ao CHN nos sistemas de manejo milho/braquiária e milho solteiro, e mostrou valores de COP semelhantes àqueles obtidos pelo CHN nos sistemas de manejo milho solteiro e milho/braquiária. Valores de COT, COP e COM obtidos pela metodologia de Mebius modificado foram semelhantes àqueles obtidos pelo analisador elementar, para todos os sitemas de manejo. O COP, quando avaliado pelas metodologias de Mebius e CHN, mostrou-se significativamente mais sensível às alterações de manejo em relação ao COT e COM.
Soil organic matter alterations can be better understood when it is fractionated for the identification of its labile (> 53 µm) and stable (< 53 µm) fractions. Because of the increasing and important use of soil organic matter fractionation techniques, it becomes necessary to evaluate if the carbon recovery in these fractions is influenced by methods used for its determination. The aim of this work was to evaluate the content of organic carbon in soil organic matter fractions, by three different methods, under maize-grass intercropping and sole maize in a dystrophic Red-Yellow Latosol (Oxisol). The following carbon contents were determined from the fractions: (i) of the particulate soil organic matter (POM) and from that (ii) of mineral-associated soil organic matter (MOC). The content of total organic carbon (TOC) was also determined in the non-fractionated soil organic matter. Three methods for the determination of soil organic carbon were used: (i) Walkley & Black modified, (ii) Mebius modified, and (iii) dry combustion in a CHN elemental analyzer. The experimental design was a randomized block design with three replications. Soil samples were collected from three management systems: (i) sole corn; (ii) maize-Panicum maximum intercropping; and (iii) maize- Brachiaria humidicola intercropping. Soil samples were collected from the depth of 0 5 cm. Lower TOC were found in the Walkley & Black method in relation to the CHN elemental analyzer, under the management systems sole maize and maize-Panicum maximum intercropping. However, the Walkley & Black method showed similar POM in relation to CHN analyzer method, under the management systems sole maize and maize-Brachiaria humidicola intercropping. TOC, POM and MOC obtained from the Mebius modified method were similar to those obtained from the elemental analyzer, under the three management systems. The POM when evaluated by the Mebius and CHN methods, showed to be more sensible to soil management alterations.
Assuntos
Carbono , Agricultura Sustentável , Matéria Orgânica , Fracionamento QuímicoRESUMO
Dental caries is considered a multi-factorial, infectious, chronic, localized, post-eruptive, transmissible disease that leads to the destruction of dental hard tissue. The recognition of Streptococcus mutans as the major bacterial species involved in dental caries has led to the implementation of prevention and control measures for eliminating or reducing it in oral cavity. The main goal of research on medicinal plants is the search for substances or compounds with antimicrobial activity. The aim of this study was to evaluate the antimicrobial activity of fractions obtained by two methods from Isertia laevis against S. mutans and S. sobrinus. The plant material was collected in Medina (Colombia), at an elevation of 550 meters above sea level. From the ethanol extract of leaves of I. laevis, fractions were obtained by two methods: extraction by column vacuum chromatography (CVC) and extraction by continuous liquid / liquid partitioning (CLLP). The evaluation of the antimicrobial activity of fractions against S. mutans and S. sobrinus was performed by well diffusion and bioautography assays. From the CVC technique, only the methanol and methanol-dichloromethane fractions showed activity against S. mutans and S. sobrinus, with a minimum inhibitory concentration of 2 mg/well. From the CLLP technique, only the dichloromethane fraction showed activity against both microorganisms, with a minimum inhibitory concentration of 1 mg/well. Compounds C1 and C2 were isolated from the three active fractions, and showed a minimum inhibitory concentration of 0.4 mg/well for S. mutans and S. sobrinus, with zones of inhibition measuring 6.5 and 6.2 mm, respectively. In conclusion: 1) the three active fractions of I. laevis showed activity against S. mutans and S. sobrinus, 2) compounds C1 and C2 were present equally in the three active fractions showing activity against the two bacteria, 3) compounds C1 and C2 may be triterpenoid and/or steroidal saponin structures, and 4) the two extraction methods lead equally to obtaining the active fractions.
La caries dental es considerada una enfermedad infecciosa multifactorial, cronica, localizada, poseruptiva y transmisible que conlleva a la destruccion del tejido dental duro. El claro reconocimiento de Streptococcus mutans como la principal especie bacteriana implicada en caries dental, ha conducido a la implementacion de medidas de prevencion y control para la eliminacion o disminucion de este microorganismo en cavidad oral. El objetivo fundamental de la investigacion en plantas medicinales, es la busqueda de sustancias o compuestos con actividad antimicrobiana para ser utilizadas en el control o prevencion de enfermedades infecciosas. En este sentido, en salud bucal muchas sustancias obtenidas de plantas han mostrado actividad antimicrobiana. El objetivo de este estudio fue evaluar la actividad antimicrobiana de fracciones obtenidas de la planta Isertia laevis mediante dos metodologias contra S. mutans y S. sobrinus. El material vegetal se colecto en el municipio de Medina (Cundinamarca- Colombia) situado a una altura de 550 metros sobre el nivel del mar. A partir del extracto etanolico de hojas de I. laevis se obtuvieron fracciones mediante dos metodologias, extraccion por cromatografia en columna al vacio (CCV) y extraccion por fraccionamiento liquido/liquido continuo (FLLC). La evaluacion de la actividad antimicrobiana de las fracciones frente a S. mutans y S. sobrinus se realizo por el metodo de difusion en pozo y bioautografico. De las fracciones obtenidas por CCV, solamente las fracciones metanol y metanol-diclorometano presentaron actividad antimicrobiana sobre S. mutans y S. sobrinus, con una concentracion minima inhibitoria de 2 mg/pozo. De las fracciones obtenidas por FLLC solamente la fraccion diclorometano presento actividad antimicrobiana sobre S. mutans y S. sobrinus, con una concentracion minima inhibitoria de 1 mg/pozo. De las 3 fracciones activas se aislaron los compuestos C1 y C2, que presentaron una concentracion minima inhibitoria de 0.4 mg/pozo tanto para S. mutans como para S. sobrinus con halos de inhibicion, respectivamente, de 6.5 y 6.2 mm. En conclusion, 1. Las fracciones metanol y diclorometano obtenidas por CCV y la fraccion diclorometano obtenida por FLLC de hojas de I. laevis presentaron actividad antimicrobiana sobre S. mutans y S. sobrinus; 2. Los compuestos C1 y C2 presentes por igual en las tres fracciones activas tuvieron accion inhibitoria sobre las dos bacterias en evaluacion; 3. Las pruebas quimicas cualitativas para los compuestos C1 y C2 indican que posiblemente corresponden a estructuras de saponinas triterpenicas y/o esteroidales; y 4. Las dos metodologias de extraccion conducen por igual a la obtencion de las fracciones activas.
Assuntos
Humanos , Streptococcus mutans/efeitos dos fármacos , Extratos Vegetais/farmacologia , Cariostáticos/farmacologia , Streptococcus sobrinus/efeitos dos fármacos , Rubiaceae , Solventes/química , Terpenos/química , Vácuo , Extratos Vegetais/química , Testes de Sensibilidade Microbiana , Cromatografia/métodos , Cromatografia Líquida , Imunodifusão , Etanol/química , Metanol/química , Fracionamento Químico , Cloreto de Metileno/químicaRESUMO
Natural essential oils are used extensively in fragrances, flavorants, and in the food and pharmaceutical industries. During hydrodistillation, a part of the essential oil becomes dissolved in the condensate and lost as this water is discarded. In this study, carvone and limonene content recovered from hydrodistillation waste water of caraway fruit were quantified using two methods for recovering dissolved aromatic molecules from condensate water: extraction through distillation and extraction by means of a solvent. This allows for the conservation of useful molecules which are typically discarded with the waste water produced during the distillation process. The objective of this study was to quantify the carvone and limonene content recoverable from waste water derived from the distillation of caraway essential oil. The well-known Clevenger method and a simpler, more practical technique employing cyclohexane as a solvent were employed to determine the recoverable content of aromatic molecules from the hydrosol. The chemical compositions of the respective recovered extracts were compared with those of the primary oils to analyze the efficacy of these methods. Recovered extract accounted for 10 to 40 percent of the total oil yield. The limonene and carvone molecules recovered using these methods were quantified through gas chromatography in order to characterize the composition of the secondary extract produced.
Los aceites esenciales naturales se utilizan ampliamente en las fragancias, saborizantes, y en la industria alimentaria y farmacéutica. Durante la hidrodestilación, una parte del aceite esencial se disuelve en el condensado y se pierde como agua de descarga. En este estudio, el contenido de carvona y limoneno recuperados del agua de desecho de la hidrodestilación de la fruta de alcaravea se cuantificaron utilizando dos métodos para recuperar las moléculas aromáticas disueltas en el agua condensada: extracción a través de la destilación y la extracción con un disolvente. Esto permite la conservación de las moléculas útiles que normalmente son desechadas con las aguas residuales producidas durante el proceso de destilación. El objetivo de este estudio fue cuantificar el contenido de carvona y limoneno recuperable de las aguas residuales procedente de la destilación del aceite esencial de alcaravea. El conocido método de Clevenger y una técnica sencilla y práctica que emplea ciclohexano como disolvente fueron utilizadas para determinar el contenido de moléculas aromáticas contenidas en el hidrosol. La composición química de los extractos recuperados fue comparada con los aceites primarios para analizar la eficacia de estos métodos. El extracto recuperado representa del 10 al 40 por ciento del contenido total de aceite esencial. Las moléculas de limoneno y carvona recuperadas mediante estos métodos se cuantificaron mediante cromatografía de gases con el fin de caracterizar la composición del extracto secundario.
Assuntos
Óleos Voláteis/isolamento & purificação , Óleos Voláteis/química , Carum , Cicloexanos/análise , Fracionamento Químico/métodos , Monoterpenos/análise , Cromatografia Gasosa , Destilação , Terpenos/análiseRESUMO
Las mordeduras producidas por serpientes venenosas son un serio problema médico en varias regiones del mundo y sobre las cuales los sistemas de salud actúan en diferentes grados en lo referente a tratamiento y prevención. Sin embargo, el tratamiento de las mordeduras de serpientes venenosas en animales domésticos puede resultar difícil por diversos motivos, siendo uno de estos la baja oferta o ausencia de antivenenos para uso veterinario. Las presiones comerciales en la industria farmacéutica han llevado a una reducción en la producción de antivenenos en varias partes del mundo, su disponibilidad es, a veces, bastante limitada y en algunos casos, son imposibles de conseguir. En este trabajo, inmunizamos caballos con veneno de serpientes Sudamericanas para obtener el plasma hiperinmune que fue procesado para obtener IgG entera o fragmentos F(ab´)2 usando dos métodos convencionales (fraccionamiento por ácido caprílico o doble precipitación salina y digestión con pepsina). Los antivenenos así obtenidos fueron probados en sus características bioquímicas e inmunoquímicas, así como en su potencia neutralizante. El SDS-PAGE de los antivenenos mostró bandas en el orden de los 150 y 100 kDa en los antivenenos conteniendo IgG entera o fragmentos F(ab´)2, respectivamente. La presencia de albúmina o contaminantes de alto o bajo peso molecular no fue detectada en ninguna de las preparaciones. No se observaron diferencias importantes en la potencia neutralizante de los antivenenos, aunque el costo de producción fue mucho más bajo en la obtención de IgG completa. A partir de esto, se sugiere que los bajos costos de producción en la obtención de antivenenos de IgG entera para uso veterinario, hacen a esta tecnología adecuada y rentable cuando la producción de F(ab´)2 no es posible.
Bites by venomous snakes are a serious medical problem in several regions of the world, on which the different health systems act with different modalities. Nevertheless, the treatment of venomous snakebites in domestic animals can turn difficult due several problems among which, the conspicuous, is the low availability or lack of antivenoms for veterinary use. As commercial pressures on the pharmaceutical industry have led to a reduction in the production of antivenins in several parts of the world, their availability is sometimes rather limited and sometimes these products are impossible to obtain. In this work, we immunized horses with venom of South American vipers to obtain hyperimmune plasma. The plasma was processed to separate whole IgG of F(ab´)2 fragments using two conventional methods (caprylic acid fractionation or double saline precipitation and pepsin digestion). The obtained antivenins were tested for their biochemical and immunochemical characteristics and neutralizing potency. The SDS-PAGE of the antivenins showed, in the processed antivenin, bands in the order of 150 and 100 kDa in the whole IgG or F(ab´)2 fragments, respectively. The presence of albumin or contaminants of high or low molecular weight was not detected in any of the preparations. No important differences were observed in the neutralizing potency of the antivenins, although production cost was very low with the method used to obtain pure IgG. The low production cost makes the production of antivenins for veterinary use profitable when the production of F(ab´)2 fragments is not possible.
Assuntos
Animais , Antivenenos/uso terapêutico , Fragmentos Fab das Imunoglobulinas/farmacologia , Mordeduras de Serpentes , Venenos de Serpentes , Caprilatos , Fracionamento Químico/métodos , CavalosRESUMO
Comparative studies on fatty acid and protein composition of the endosperm and embryo of palmito (Euterpe edulis Martius) were conducted using gas-liquid chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. On a dry weight basis, the embryo contained extremely lower amounts of lipids and proteins than did the endosperm, which was associated with the scarce lipid and protein bodies previously reported in axis and cotyledon. The fatty acid composition also exhibited differences between both tissues: (I) the fatty acid diversity was greater in embryo than in endosperm; (II) embryo and endosperm contained predominantly linoleic, palmitic, oleic and stearic acids even though the relative values were different for each tissue. As compared to other palm species, the higher fatty acid unsaturation in Euterpe edulis seed could be involved in the previously reported short longevity and recalcitrant behavior during storage. Proteins of both tissues were heterogeneous in molecular mass. Some proteins were tissue-specific, but other were common, among them a highly glycosylated protein which migrated at about 55 kDa. We hypothesize that the latter, also reported in all previously studied palm species, is one of the proteins characterizing the Arecaceae family.
Assuntos
Ácidos Graxos/análise , Arecaceae/metabolismo , Frutas/química , Proteínas de Plantas/metabolismo , Sementes/metabolismo , Sementes/química , Fracionamento Químico , Eletroforese em Gel de Poliacrilamida , Metabolismo dos LipídeosRESUMO
El hecho de reconocer a S. mutans como el microorganismos más importante en la iniciación de la caries dental conduce a diseñar medidas dirigidas hacia la eliminación o disminución de éste en cavidad oral. Además, con el elevado índice de resistencia a los antibióticos, existe un considerable interés por estudiar los efectos de compuestos naturales obtenidos de plantas sobre microorganismos de importancia en cavidad oral. Isertia, género de la familia Rubiaceae, se distribuye desde Centroamérica hasta el norte de Sudamérica; en Colombia se conocen siete especies de este género en diferente regiones del país. Los objetivos de este estudio fueron evaluar la actividad antimicrobiana de extractos, fracciones y subfracciones obtenidas a partir de hojas de Isertia laevis sobre 18 cepas bacterianas pertenecientes a los géneros Streptococcus, Lactobacillus y Actinomyces, asociadas a la caries dental y a otras patologías bucales. La evaluación antimicrobiana se realizó de acuerdo al método de difusión en pozo contra las siguientes bacterias: S. mutans ATCC 25175, S. mutans ATCC 31989, S. mitis 804 NCTC 3165, S. salivarius NCTC 8606, S. mutans 10923, L. acidophilus 903 ATCC 4365, S. sobrinus, S. rattus FA 1 (G), S. mutans C67-1, S. cricetus AHT, S. mutans ingbritt, L. plantarum 748, L. casei 475, L. brevis, A. odontolyticus, S. mutans 35FS3m S, nytabs 35FS1 t S. mutans 29FS2. El extracto en etanol y la fracción MeOH: H20 mostraron actividad inhibitoria a 20 y 10mg sobre todas las cepas. A partir de la fracción en MeOH:H20 se obtuvieron diferentes subfracciones. Hasta la actualidad, la especie A. laevis no había sido objeto de estudio químico ni microbiológico, por lo que representa, de acuerdo a los resultados obtenidos, una promisoria fuente, en la que se deben seguir estudiando los efectos antimicrobianos en modelos más cercanos al evento infeccioso in vivo y los blancos de acción.
Assuntos
Cárie Dentária/tratamento farmacológico , Fitoterapia , Preparações de Plantas/uso terapêutico , Rubiaceae , Actinomyces , Fracionamento Químico , Cárie Dentária/microbiologia , Estudo de Avaliação , Espectrofotometria/métodos , Lactobacillus , Streptococcus mutansRESUMO
Finding a new microbial source of transglutaminase (MTGase) and the study of the medium composition for MTGase production were the goals of this work. A total of 200 actinomycete-like strains were isolated from Brazilian soil samples and two of them named T10b and P20 were selected based on their ability to produce 0.15 U.mL-1 and 0.25 U.mL-1 of MTGase, respectively. Strain P20 was chosen to continue the study and was identified as Streptomyces sp. In order to optimize the MTGase activity, modifications of the usual media composition described for enzyme production were tested. The strategy adopted was: (1) screening experiment for the best carbon and nitrogen sources; (2) fractional factorial design (FFD) to elucidate the key ingredients in the media (the results indicated that the soybean flour, peptone, KH2PO4 and MgSO4.7H2O had a significant effect on MTGase) production and (3) central composite design (CCD) to optimize the concentration of the key components. The experimental results were fitted to a second-order polynomial model at the 95 percent level of significance (P < 0.05). Under the proposed optimized conditions, the model predicted a MTGase activity of 1.37 U.mL-1, very closely matching the experimental activity of 1.4 U.mL-1.
Assuntos
Streptomyces/enzimologia , Transglutaminases/química , Análise de Variância , Actinobacteria/isolamento & purificação , Fracionamento Químico , Meios de Cultura , Fermentação , Modelos Biológicos , ViscosidadeRESUMO
Avaliaram-se as frações dos compostos nitrogenados dos capins setária (Setaria anceps Stapf), hemarthria (Hemarthria altissima [Poir] Stapf. & Hubbard), angola (Brachiaria purpurascens [Raddi] Henr.) e acroceres (Acroceras macrum Stapf.) adubados com 0, 100, 200, 300 e 400kg de nitrogênio/ha e colhidos aos 28, 42, 56 e 70 dias de idade, utilizando-se análise de fatores. Após redução e avaliação da variação conjunta total das variáveis, optou-se pela adoção de três fatores, os quais englobaram 85,6 por cento da variação total, em que o primeiro fator associou-se intimamente aos compostos nitrogenados ligados à parede celular (frações B3 e C), o segundo fator associou-se aos compostos nitrogenados protéicos citoplasmáticos de rápida e intermediária degradação (fração B1+B2), e o terceiro fator associou-se aos compostos nitrogenados não-protéicos (NNP) (fração A). As frações dos compostos nitrogenados das forrageiras foram influenciadas pela idade de corte, e o avanço da idade das plantas resultou em redução dos teores de NNP. A adubação nitrogenada contribuiu para a elevação dos teores das frações de NNP e B1+B2, e as frações associadas à parede celular não apresentaram respostas evidentes quanto à adubação nitrogenada.
The effects of levels of nitrogen fertilizer 0, 100, 200, 300 and 400kg of nitrogen/ha, and cutting ages at 28, 42, 56 and 70 days on nitrogenous compounds of tropical grasses, Setaria grass (Setaria anceps Stapf), Limpo grass (Hemarthria altissima [Poir] Stapf. & Hubbard), California grass (Brachiaria purpurascens [Raddi] Henr.) and Nilo grass (Acroceras macrum Stpaf) were evaluated, using factor analysis. After reduction and evaluation of the total variation, three selected factors accounted for 85.60 percent of the total variation, the first factor was strongly associated with nitrogenous compounds of cell wall (fractions B3 and C), the second factor, was related to citoplasmatic protein and to fast and intermediary degradations rates (fraction B1+B2), while the third factor was related to nonproteic nitrogenous compounds (NPN) (fraction A). Cutting age affected nitrogenous compounds fractions of the grasses. The increase in cutting age resulted in a reduction of NPN fraction scores. The level of nitrogen fertilizer resulted in increases of NPN and B1+B2 fractions scores, however, no evident responses in other fractions were related to nitrogen fertilizer supply.
Assuntos
Compostos de Nitrogênio/administração & dosagem , Poaceae/química , Fracionamento QuímicoRESUMO
Se presenta el estudio de la actividad antibacteriana de la especie vegetal Diplostephium tolimense Cuatrec. (Asteraceae), colectada en el páramo del Nevado del Tolima, la cual ha tenido pocos estudios químicos y farmacológicos, al igual que muchas otras especies de este ecosistema. A partir de un extracto etanólico crudo, y mediante partición biodirigida, se obtiene la fracción más simple de mayor actividad frente a Staphylococcus aureus (ATCC 25923). Paralelamente se realizan análisis fitoquímicos a las porciones de mayor bioactividad obtenidas durante todo el proceso. Se observa que la acción antibacteriana se incrementa al aumentar la simplicidad química del extracto, y que terpenos y flavonoides parecen estar relacionados con la acción revelada. Se trata del primer reporte, hasta la fecha conocido, de actividad antibacteriana in vitro para D. tolimense
Assuntos
Plantas , Terpenos , Extratos Vegetais , Fracionamento QuímicoRESUMO
Different immunomodulatory activities present in Trichilia glabra (TG) leaf extracts have already been described. Particularly, chloroform-methanol extracts were responsible for an in-vivo anti-inflammatory effect. The effect of such extracts on the infectivity of enveloped and naked viruses were investigated. Methanolic fraction extracts were active against herpes simplex virus type 1 (HSV-1) and vesicular stomatitis virus (VSV), while no activity against poliovirus type 3 was observed. VSV was slightly more affected than HSV-1: 2.8 log10 reduction in VSV titer against 2.4 log10reduction in HSV-1 titer when 0.25 mg/ml F2 fraction was tested and a reduction of 2.7 log10in VSV virus titer and of 1.5 log10in HSV-1 virus titer was observed when 0.25 mg/ml F3 fraction was tested. Results obtained in this work suggest a potential pharmaceutical use of TG extract components.
Previamente se han descripto distintas actividades inmunomoduladoras, presentes en extractos de hojas de Trichilia glabra (TG). En particular, se ha demostrado una actividad antiinflamatoria presente en extractos metanólicos. En este trabajo se investigó la actividad virucida de dichos extractos sobre virus envueltos y desnudos. Distintos extractos metanólicos han inactivado en forma moderada los virus herpes simplex tipo 1 (HSV-1) y el virus de la estomatitis vesicular (VSV), mientras no evidenciaron actividad sobre poliovirus tipo 3. VSV resultó algo mas afectado que HSV-1: se observó una reducción en el título viral de 2,8 log10para VSV y de 2,4 log10para HSV-1 cuando se uso una concentración de 0,25 mg/ml de la fracción F2 y una reducción de 2,7 log10para VSV y de 1,5 log 10para HSV-1 cuando se usó una concentración de 0,25 mg/ml de la fracción F3. Los resultados obtenidos en este trabajo, sugieren un potencial uso farmacéutico de los componentes presentes en los extractos de TG.
Assuntos
Animais , Antivirais/isolamento & purificação , Meliaceae/química , Extratos Vegetais/farmacologia , Folhas de Planta/química , Antivirais/farmacologia , Chlorocebus aethiops , Fracionamento Químico , Clorofórmio , Herpesvirus Humano 1/efeitos dos fármacos , Metanol , Extratos Vegetais/isolamento & purificação , Poliovirus/efeitos dos fármacos , Células Vero , Vírus da Estomatite Vesicular Indiana/efeitos dos fármacosRESUMO
A radioterapia é uma opção terapêutica padrão para o tratamento radical do câncer de próstata localizado. Umamelhora nos índices terapêuticos, através da elevação da dose total do tratamento pelas irradiações, tem sidoexaustivamente pesquisada nos últimos anos, com o auxílio de modernos e mais precisos sistemas de planejamentoe de tratamento. Recentemente, um interesse renovado sobre o uso do hipofracionamento para o tratamento docâncer de próstata pela radioterapia está sendo observado. Tal mudança de estratégia está relacionada ao novoconceito radiobiológico de que os tumores prostáticos possuem um índice α/β muito mais baixo do que previamenteestimado. O hipofracionamento, portanto, parece ser uma opção terapêutica interessante para este tumor, tendoem vista a possibilidade de melhora do índice terapêutico com uma alteração na dose diária de radioterapia. Estetrabalho avalia criticamente o valor do hipofracionamento nos tumores prostáticos malignos.