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1.
Braz. j. biol ; 84: e255080, 2024. tab, graf, ilus
Article in English | LILACS, VETINDEX | ID: biblio-1364503

ABSTRACT

In the current context of emerging drug-resistant fungal pathogens such as Candida albicans and Candida parapsilosis, discovery of new antifungal agents is an urgent matter. This research aimed to evaluate the antifungal potential of 2-chloro-N-phenylacetamide against fluconazole-resistant clinical strains of C. albicans and C. parapsilosis. The antifungal activity of 2-chloro-N-phenylacetamide was evaluated in vitro by the determination of the minimum inhibitory concentration (MIC), minimum fungicidal concentration (MFC), inhibition of biofilm formation and its rupture, sorbitol and ergosterol assays, and association between this molecule and common antifungal drugs, amphotericin B and fluconazole. The test product inhibited all strains of C. albicans and C. parapsilosis, with a MIC ranging from 128 to 256 µg.mL-1, and a MFC of 512-1,024 µg.mL-1. It also inhibited up to 92% of biofilm formation and rupture of up to 87% of preformed biofilm. 2-chloro-N-phenylacetamide did not promote antifungal activity through binding to cellular membrane ergosterol nor it damages the fungal cell wall. Antagonism was observed when combining this substance with amphotericin B and fluconazole. The substance exhibited significant antifungal activity by inhibiting both planktonic cells and biofilm of fluconazole-resistant strains. Its combination with other antifungals should be avoided and its mechanism of action remains to be established.


No atual contexto de patógenos fúngicos resistentes emergentes tais como Candida albicans e Candida parapsilosis, a descoberta de novos agentes antifúngicos é uma questão urgente. Esta pesquisa teve como objetivo avaliar o potencial antifúngico da 2-cloro-N-fenilacetamida contra cepas clínicas de C. albicans e C. parapsilosis resistentes a fluconazol. A atividade antifúngica da substância foi avaliada in vitro através da determinação da concentração inibitória mínima (CIM), concentração fungicida mínima (CFM), ruptura e inibição da formação de biofilme, ensaios de sorbitol e ergosterol, e associação entre esta molécula e antifúngicos comuns, anfotericina B e fluconazol. O produto teste inibiu todas as cepas de C. albicans e C. parapsilosis, com uma CIM variando de 128 a 256 µg.mL-1, e uma CFM de 512-1,024 µg.mL-1. Também inibiu até 92% da formação de biofilme e causou a ruptura de até 87% de biofilme pré-formado. A 2-cloro-N-fenilacetamida não promoveu atividade antifúngica pela ligação ao ergosterol da membrana celular fúngica, tampouco danificou a parede celular. Antagonismo foi observado ao combinar esta substância com anfotericina B e fluconazol. A substância exibiu atividade antifúngica significativa ao inibir tanto as células planctônicas quanto o biofilme das cepas resistentes ao fluconazol. Sua combinação com outros antifúngicos deve ser evitada e seu mecanismo de ação deve ser estabelecido.


Subject(s)
In Vitro Techniques , Candida albicans , Fluconazole , Candida parapsilosis , Antifungal Agents
2.
Acta odontol. Colomb. (En linea) ; 12(2): 61-77, Jul-Dec. 2022. tab, graf
Article in Spanish | LILACS | ID: biblio-1397171

ABSTRACT

Objetivo: establecer los parámetros para la evaluación visual e instrumental del color dental en estudios in-vitro a partir de la literatura científca publicada entre 2015 y 2021. Métodos: se realizó la búsqueda en las bases de datos: PubMed, Web of Science, Science Direct, Scopus, Scielo y Lilacs; también en el motor de búsqueda Google Académico y las bibliotecas de las editoriales Wiley y Springer. Las palabras clave utilizadas fueron tooth, color, in-vitro, color perception, shade matching, thresholds, appearance, surrounding, "CIELAB" y "CIEDE2000". Teniendo en cuenta los criterios de elegibilidad, se seleccionaron los estudios de acuerdo al título, resumen y texto completo. Resultados: la búsqueda arrojó un total de 37 publicaciones que se agruparon en tres tópicos: 1. toma de color visual: condiciones ambientales, observadores y nivelación; 2. toma de color instrumental: instrumentos; y 3. procesamiento de datos: cálculo de la diferencia de color y umbrales de perceptibilidad (PT) y aceptabilidad (AT). Conclusiones: los aspectos más importantes en la evaluación visual son la iluminación, el ambiente para registro (sitio, entorno y fondo alrededor de la muestra), las condiciones geométricas de visualización, los observadores y el uso de guías. En la evaluación instrumental es relevante elegir el aparato apropiado de acuerdo con su precisión y reproducibilidad, como los espectroradiómetros y los espectrofotómetros de uso clínico. Se presenta el procesamiento de datos para establecer las variaciones de cada coordenada, las diferencias de color (ΔE): CIELAB y CIEDE2000, los umbrales y los lineamientos.


Objective: To establish the parameters for the visual and instrumental evaluation of tooth color in in-vitro studies based on the scientifc literature published between 2015 and 2021. Methods: The search was carried out in the databases of PubMed, Web of Science, Science Direct, Scopus, Scielo, Lilacs; search engine Google Scholar and publishers' library of Wiley and Scielo, using the keywords "tooth", "color", "in vitro", "color perception", "shade matching", "thresholds", "appearance", "surrounding", "CIELAB", and "CIEDE2000". The literature was selected according to the title, abstract and full text taking into the eligibility criteria. Results: It yielded a total of 37 publications, which were grouped into three topics: 1. visual color acquisition: environmental conditions for color acquisition, observers and levelling. 2. instrumental color sampling: instruments. 3. Data processing: Calculation of color diference and perception thresholds (PT) and acceptability thresholds (AT). Conclusions: The most important aspects in the visual assessment are lighting, the environment for color registration (site, environment and background around the sample), the geometric conditions of visualization, the observers and the use of guides. Regarding the instrumental assessment of color, the appropriate devices must be chosen according to its precision and reproducibility, being the spectrophotometers and spectroradiometers the most precise ones. It is presented how the data processing is carried out to establish the variations of each coordinate, the color diferences (ΔE): CIELAB and CIEDE2000, thresholds and guidelines.


Subject(s)
Tooth , Color Perception , In Vitro Techniques , Differential Threshold
3.
Rev. Círc. Argent. Odontol ; 80(231): 6-13, jul. 2022. ilus, tab, graf
Article in Spanish | LILACS | ID: biblio-1391619

ABSTRACT

Este trabajo tuvo como objetivo conocer la fiabilidad de la impresora 3D (i3D) aditiva por Matriz de Proceso Digital de Luz (MDLP) Hellbot modelo Apolo®, a través de verificar la congruencia dimensional entre las mallas de modelos impresos (MMi) y su correspondiente archivo digital de origen (MMo), obtenido del software de planificación ortodontica Orchestrate 3D® (O3D). Para determinar su uso en odontología y sus posibilidades clínicas, fue comparada entre cinco i3D de manufactura aditiva, dos DLP, dos por estereolitografía (SLA) y una por Depósito de Material Fundido (FDM). La elección de las cinco i3D se fundamentó en su valor de mercado, intentando abarcar la mayor diversidad argentina disponible. Veinte modelos fueron impresos con cada i3D y escaneados con Escáner Intraoral (IOS) Carestream modelo 3600® (Cs3600). Las 120 MMi fueron importadas dentro del programa de ingeniería inversa Geomagic® Control X® (Cx) para su análisis 3D, consistiendo en la superposición de MMo con cada una de las MMi. Luego, una evaluación cualitativa de la desviación entre la MMi y MMo fue realizada. Un análisis estadístico cuidadoso fue realizado obteniendo como resultado comparaciones en 3d y 2d. Las coincidencias metrológicas en la superposición tridimensional permitieron un análisis exhaustivo y fácilmente reconocible a través de mapas colorimétricos. En el análisis bidimensional se plantearon planos referenciados dentariamente desde la MMo, para hacer coincidir las mediciones desde el mismo punto de partida dentaria. Los resultados fueron satisfactorios y muy alentadores. Las probabilidades de obtener rangos de variabilidad equivalentes a +/- 50µm fueron de un 40,35 % y de +/- 100µm un 71,04 %. Por lo tanto, te- niendo en cuenta las exigencias de congruencia dimensional clínicas de precisión y exactitud a las cuales es sometida nuestra profesión odontológica, se evitan problemas clínicos arrastrados por los errores dimensionales en la manufactura (Cam) (AU)


The objective of this study was to determine the reliability of the Hellbot Apollo® model additive 3D printer (i3D) by Matrix Digital Light Processing (MDLP) by verifying the dimensional congruence between the printed model meshes (MMi) and their corresponding digital source file (MMo), obtained from the Orchestrate 3D® (O3D) orthodontic planning software. A comparison was made between five i3D of additive manufacturing, two DLP, two by stereolithography (SLA), and one by Fused Material Deposition (FDM), to determine its use in dentistry and its clinical possibilities. The choice of the five i3D was based on their market value, trying to cover most of the Argentinean diversity available. Twenty models were printed with each i3D and scanned with Carestream Intraoral Scanner (IOS) model 3600® (Cs3600). The 120 MMi were imported into the reverse engineering program Geomagic® Control X® (Cx) for 3D analysis, consisting of overlaying MMo with each MMi. Then, a qualitative evaluation of the deviation between MMi and MMo. Also, a careful statistical analysis was performed, resulting in 3d and 2d comparisons. Metrological coincidences in three-dimensional overlay allowed a comprehensive and easily recognizable analysis through colorimetric maps. In the two-dimensional analysis, dentally referenced planes were proposed from the MMo, to match the measurements from the same dental starting point. The results were satisfactory and very encouraging. The probabilities of obtaining ranges of variability equivalent to +/- 50µm were 40.35 % and +/- 100µm 71.04 %. Therefore, considering the demands of clinical dimensional congruence, precision, and accuracy to which our dental profession it is subjected, clinical problems caused by dimensional errors in manufacturing (Cam) are avoided (AU)


Subject(s)
Models, Dental , Printing, Three-Dimensional , Stereolithography , Orthodontics/methods , In Vitro Techniques , Algorithms , Software , Image Interpretation, Computer-Assisted/methods , Data Interpretation, Statistical , Evaluation Studies as Topic
4.
Rev. ADM ; 79(3): 146-151, mayo-jun. 2022. ilus, tab
Article in Spanish | LILACS | ID: biblio-1378372

ABSTRACT

Introducción: La finalidad del tratamiento de conductos es conseguir la máxima desinfección, conformación y sellado tridimensional. Objetivo: Comparar la capacidad del sellado apical entre dos técnicas de obturación en conductos mesiales de molares inferiores con limas fracturadas en tercio apical. Material y métodos: Se utilizaron 60 raíces mesiales de molares inferiores, instrumentados con Protaper Universal, se desgastó 4 milímetros la parte activa del instrumento y se fracturó intencionalmente en tercio apical. Se formaron dos grupos de 30 raíces mesiales (n = 30) cada uno y se utilizaron dos técnicas de obturación: grupo 1: condensación lateral clásica y grupo 2: Obtura II. Las muestras se sumergieron en tubos de ensayo y en su interior contenían 5 mL de tinta china, se diafanizaron y observaron con un microscopio estereoscópico (LEICA, EZ4D) a 35x para medir la penetración de tinta china dentro del conducto radicular. Resultados: Se encontró una mayor microfiltración apical con suficiente evidencia estadística en el grupo de Obtura II comparado con el grupo de condensación lateral clásica (p < 0.002). Conclusiones: Ambos grupos presentaron microfiltración apical; sin embargo, en el grupo que se utilizó el sistema de obturación termoplastificada Obtura II se detectó mayor filtración apical comparada con el grupo de condensación lateral clásica (AU)


Introduction: The purpose of root canal treatment is to achieve maximum disinfection, shaping and three-dimensional sealing. Objective: To compare the apical sealing capacity between two obturation techniques in mesial canals of mandibular molars with broken files in the apical third. Material and methods: 60 mesial roots of lower molars were used, instrumented with Protaper Universal, the active part of the instrument was worn by 4 millimeters and it was intentionally broken in the apical third. Two groups of 30 mesial roots (n = 30) each were formed and two filling techniques were used: group 1: classic lateral condensation and group 2: Obtura II. The samples were immersed in test tubes and contained 5 mL of Chinese ink inside, they were clear and observed with a stereomicroscope (LEICA, EZ4D) at 35x to measure the penetration of Chinese ink into the root canal. Results: A higher apical microfiltration with sufficient statistical evidence was found in the Obtura II group compared to the classic lateral condensation group (p < 0.002). Conclusions: Both groups presented apical microfiltration, however, in the group that used the Obtura II thermoplastic obturation system, greater apical filtration was detected compared to the classic lateral condensation group (AU)


Subject(s)
Root Canal Obturation/methods , Dental High-Speed Equipment/adverse effects , Dental Leakage , In Vitro Techniques , Cross-Sectional Studies , Dental Restoration Failure , Dental Pulp Cavity/anatomy & histology , Molar
5.
Int. j. high dilution res ; 21(1): 19-19, May 6, 2022.
Article in English | LILACS, HomeoIndex | ID: biblio-1396573

ABSTRACT

Sporothrix brasiliensisis one of themost virulent zoonosis which affects animals and humans. This fungus is responsible for subcutaneous infection and its contamination is possible through trauma to the skin. Sporotrichosis is highly prevalent in feline. And Rio de Janeiro appears to have the highest occurrence of cases. Objectives: This study aims to evaluate the in vitroefficacy of Sporothrix brasiliensisbiotherapic, with and without an association to allopathic medicine commonly used in the treatment.Methodology: Conidiumcells of Sporothrix brasiliensiswill be cultured in Potato dextrose agar (PDA) for 5 to 7 days and yeast cells in Brain heart infusion (BHI) for 3 to 5 days. After incubation, the cells will be scraped with a drigalski handle and filtered using cells strainer into a tube and centrifuge for 5 minutes at 3000 RPM. The cells will be resuspended with Phosphate buffered saline (PBS), centrifuge again,and finally resuspended in PBS. After preparing the inocule, the microplates will be prepared. There will be 5groups in vitro. The first one will be the control group, only fungi. The second will be the treatment of fungi with homeopathic medicine (Sporothrix brasiliensis30DH). The third group will be the homeopathic medicine in association with itraconazole. The fourth will be the treatment with itraconazole only. And the last group will be the fungi with dynamized distilled water 30DH. Sporothrix brasiliensis30DHwill be prepared according to Brazilian Homeopathic Pharmacopeia. Results and discussions: The experiments are still in progress and the results will be analyzed through Analysis of variance (ANOVA) to determine statistically significant differences. Previous articles based on biotherapic treatments demonstrated successful results, so our research group is conducting these experiments to evaluate the effect in this model. Conclusion: Experiments will be made to verify the efficacy of biotherapic on sporotrichosis treatments.


Subject(s)
Sporotrichosis/therapy , In Vitro Techniques , Biotherapics/therapeutic use
6.
Int. j. high dilution res ; 21(1): 18-18, May 6, 2022.
Article in English | LILACS, HomeoIndex | ID: biblio-1396574

ABSTRACT

Studies have shownthat homeopathy modulates the activity of both single-and multi-celled organisms;therefore, we propose a study into the action of Arnica Montanaand S. cerevisiae fungus nosode on growth "in vitro", and on the fermentation of S. cerevisiaeon brewer's wort. Methods:250 µL of medication in 30% alcohol were placed in 5 mL of Sabouraud Broth (SB) or wort, with 20 µL of fungus ata McFarland standard of 0.5 and in a dilution of 1:100. Fungal growth was evaluated via spectrophotometry at 600 nm or a cell count in a Neubauer chamber in a kinetic of 1 to 5 days' incubation at 25ºC. The production of alcohol by the fungus was evaluated using the BRIX index in the samekinetic. 1x107fungi/mL were previously incubated with medication for 5 days and, afterwards, placed in 20 mL of fresh wort, incubated at 25ºC for 7 days and evaluated for growth and sugar consumption. Resultsand Discussion: The SB results revealed that after 2days incubation with Arnica30CH, an increase in fungal growth was observed (p<0.0001), whilewith nosode 6 and 30CH there was a reduction in growth after 2 and 5 days incubation (p<0.001). The fungi incubated with Arnica30CH exhibited increased sugar consumption after 2 and5 days incubation (p<0.05), while the nosode 30CH resulted in lower sugar consumption after 2 and 3 days incubation (p<0.05). The results for fungal growth and sugar consumption with the wort were similar to those using SB.The fungalcultures previously incubated with homeopathic medication and subsequent incubation with fresh wortindicated a loss of distinction, bothin terms of fungal growth and sugar consumption. This piece of data may suggest action by the homeopathic medication only when in contact with the cells. Conclusion: The treatment of the S. cerevisiae fungus using Arnica and the S. cerevisiae nosode produced a significant modulation in fungal growth and sugar consumption.


Subject(s)
Saccharomyces cerevisiae/metabolism , In Vitro Techniques , Fermentation , Homeopathy
7.
Int. j. high dilution res ; 21(1): 12-12, May 6, 2022.
Article in English | LILACS, HomeoIndex | ID: biblio-1396583

ABSTRACT

The method of preserving substances of natural origin should not only maintain the microbiological safety of the product but also the integrity of its therapeutic potential. Essential oils obtained from plants are complex mixtures of substances and it issuggested to keep them under refrigeration for better conservation. On the other hand, homeopathic mother tincture prepared from plant is kept at room temperature. Aim: This work aimed to evaluate if the freezing process changes the in vitro antifungal activity potential of the homeopathic preparation Aloysia polystachya1CH against Candida albicans. Methodology:The inoculum of C. albicansATCC 10231 was cultivated in culture médium Sabouroud (Himedia®), standardized on a spectrometer and distributed in a 96-well plate. Then, A. polystachya1CH was added to the wells, prepared accordingtothe Brazilian Homeopathic Pharmacopoeia (FHB, 3rd edition) from A. polystachya essencial oil. An aliquot of this homeopathic preparation was frozen and after 40 days it was submitted to the same methodology for evaluation of the antifungal activity. After incubation, the plates were read with triphenyltetrazolic (TTC) (Vetec®). Results and discussion: The results of the in vitroevaluation showed that the freezing process retained the antifungal activity of the dynamized essential oil of A. polystachya1CH against C. albicans. Conclusion: Under the conditions evaluated in this study, the freezing method presented as a viable method of conservation of dynamized plant material.


Subject(s)
In Vitro Techniques , Candida albicans , Derived Preparations , Antifungal Agents
8.
Int. j. high dilution res ; 21(2): 10-10, May 6, 2022.
Article in English | LILACS, HomeoIndex | ID: biblio-1396744

ABSTRACT

A common clinical occurrence in dogs is otitis externa caused by excessive growth of yeasts Malassezia pachydermatis, which can become chronic after wrong treatments, in which microbial resistance can occur. Homeopathic remedies can be considered a successful alternative, selecting the medicine through the similitude principle. Herein, 50 µL of a 1:1000 dilution of Malassezia pachydermatis suspension at 0.5 McFarland scale was used to seed the yeast into Sabouraud dextrose agar plates using a Drigalski spreader to proceed with colony unit counting. Before being seeded, the yeast suspensions were treated with 1% of different homeopathic treatments previously selected from a pilot study, which means Sulphur 6cH, Dolichos pruriens 6cH, and Kali carbonicum 6cH, being water, and succussed water used as controls. For comparison, a set of Sabouraud dextrose agar plates containing 1% Tween 80 was seeded in parallel. The treatments were made blind and evaluated in triplicate. Contaminated cultures were withdrawn. The number of colonies per plate was assessed, and smears were made from the cultures to classify yeast growth according to cytomorphology on ImageJ®software. The preliminary results show no significant effect of all tested medicines compared to the controls. High data variability was also observed, mainly in those cultures whose medium was prepared with Twin 80. In conclusion, at this point of the study, no evidence of the effects of the studied medicines on Malassezia pachydermatis growth in vitro could be identified. The analysis of cytomorphology is still in course.


Subject(s)
Animals , Dogs , Otitis Externa/therapy , Yeasts , Homeopathic Therapeutics , Malassezia , In Vitro Techniques
9.
Bol. latinoam. Caribe plantas med. aromát ; 21(3): 352-364, mayo 2022. ilus, tab
Article in English | LILACS | ID: biblio-1396915

ABSTRACT

The antioxidant activity and the inhibitory potential of α-amylase of lyophilized hydroethanolic extracts of Conocarpus erectus leaves obtained by ultrasonication were determined. The most potent extract was subjected to ultra-high performance liquid chromatography system equipped with mass spectrometer for metabolite identification. The identified metabolites were docked in α-glucosidase to assess their binding mode. The results revealed that 60% ethanolic extract exhibited highest ferric reducing antioxidant power (4.08 ± 0.187 mg TE/g DE) and α-amylase inhibition (IC50 58.20 ± 1.25 µg/mL. The metabolites like ellagic acid, 3-O-methyl ellagic acid, ferujol, 5, 2 ́-dihydroxy-6,7,8-trimethyl flavone and kaempferol glucoside were identified in the extract and subjected to molecular docking studies regarding α-amylase inhibition. The comparison of binding affinities revealed 3-O-methyl ellagic acid as most effective inhibitor of α-amylase with binding energy of -14.5911 kcal/mol comparable to that of acarbose (-15.7815 kcal/mol). The secondary metabolites identified in the study may be extended further for functional food development with antidiabetic properties.


Se determinó la actividad antioxidante y el potencial inhibidor de la α-amilasa de extractos hidroetanólicos liofilizados de hojas de Conocarpus erectus obtenidos por ultrasónicación. El extracto más potente se sometió a un sistema de cromatografía líquida de ultra alto rendimiento equipado con un espectrómetro de masas para la identificación de metabolitos. Los metabolitos identificados se acoplaron en α-glucosidasa para evaluar su modo de unión. Los resultados revelaron que el extracto etanólico al 60% exhibió el mayor poder antioxidante reductor férrico (4.08 ± 0.187 mg TE/g DE) e inhibición de la α-amilasa (IC50 58.20 ± 1.25 µg/mL. Los metabolitos como el ácido elágico, 3-O-metil elágico ácido, ferujol, 5, 2 ́-dihidroxi-6,7,8-trimetil flavona y kaempferol glucósido se identificaron en el extracto y se sometieron a estudios de acoplamiento molecular con respecto a la inhibición de la α-amilasa. La comparación de las afinidades de unión reveló 3-O-metil El ácido elágico como inhibidor más eficaz de la α-amilasa con una energía de unión de -14,5911 kcal/mol comparable a la de la acarbosa (-15,7815 kcal/mol). Los metabolitos secundarios identificados en el estudio pueden ampliarse aún más para el desarrollo funcional de alimentos con propiedades antidiabéticas.


Subject(s)
Plant Extracts/chemistry , alpha-Amylases/antagonists & inhibitors , Myrtales/chemistry , Antioxidants/chemistry , Benzopyrans/analysis , In Vitro Techniques , Plant Extracts/pharmacology , Plant Leaves/chemistry , Molecular Docking Simulation , Antioxidants/pharmacology
10.
Arch. latinoam. nutr ; 72(1): 11-22, mar. 2022. tab, graf
Article in Spanish | LILACS, LIVECS | ID: biblio-1368344

ABSTRACT

El arándano (Vaccinium corymbosum L.) posee un alto contenido de compuestos fenólicos los cuales han sido estudiados principalmente por su actividad antioxidante, antiobesogénica, antiinflamatoria, entre otras. Objetivo. Evaluar el efecto de la digestión gastrointestinal in vitro sobre la bioaccesibilidad de compuestos fenólicos y actividad antioxidante de una formulación nutracéutica de arándano (cápsula), comparado con arándano fresco y polvo. Materiales y métodos. Se obtuvieron extractos metanólicos de muestras de arándano fresco y liofilizado y se determinó su contenido de fenoles, flavonoides y antocianinas totales, así como también actividad antioxidante. Se llevó a cabo un ensayo de simulación de digestión gastrointestinal para evaluar la bioaccesibilidad de los compuestos fenólicos presentes en las muestras. Resultados. Los resultados mostraron que la digestión gástrica de arándano en polvo y en cápsula promovió una mayor bioaccesibilidad de fenoles (42% y 40%), flavonoides (52% y 33%) y antocianinas (45% y 40%) comparado con digestos de arándano fresco. Posterior a la digestión intestinal, la bioaccesibilidad de fenoles (63%) y flavonoides (67%) fue mayor en la cápsula de arándano comparada con su contraparte arándano en polvo. Las condiciones de digestión intestinal afectaron negativamente la bioaccesibilidad de las antocianinas independientemente del tipo de muestra evaluada. Conclusión. Las condiciones de digestión gástrica promueven una mayor estabilidad de los compuestos fenólicos en arándano en polvo y en cápsula lo que pudiera ser relevante para el mantenimiento de un ambiente antioxidante a este nivel. Las condiciones de digestión intestinal afectaron de manera particular a los compuestos fenólicos de arándano fresco y polvo, pero no a la cápsula, lo que puede sugerir que el encapsulamiento protegió de las condiciones alcalinas a los fenoles presentes. Se sugieren estudios posteriores sobre absorción in vitro de los componentes remanentes en intestino y sus posibles efectos sobre biomarcadores de estrés oxidativo en modelos in vivo(AU)


Blueberry (Vaccinium corymbosum L.) has a high content of phenolic compounds which have been studied mainly for their antioxidant, antiobesogenic, anti-inflammatory activity, among others. Objetive. The objective of the present study was to evaluate the effect of in vitro gastrointestinal digestion on the bioaccessibility of phenolic compounds and antioxidant activity of a nutraceutical formulation of blueberry (capsule), compared to fresh and powder blueberry. Materials and methods. Methanolic extracts of fresh and lyophilized blueberry were obtained and determined its total phenols, flavonoids, anthocyanins content, as well as antioxidant activity. A gastrointestinal digestion simulation test also was carried out to assess the bioaccessibility of the phenolic compounds found in samples. Results. The results showed that gastric digestion of powder and capsule blueberry promoted greater bioaccessibility of phenols (42% and 40%), flavonoids (52% and 33%) and anthocyanins (45% and 40%), compared to fresh blueberry digests. After intestinal digestion, the bioaccessibility of phenols (63%) and flavonoids (67%) was higher in the blueberry capsule compared to its powdered blueberry counterpart. The intestinal digestion conditions negatively affected the bioaccessibility of anthocyanins regardless of the type of sample evaluated. Conclusion. Gastric digestion conditions promote greater stability of phenolic compounds in powdered and capsule blueberries, which could be relevant for the maintenance of an antioxidant environment at this level. The intestinal digestion conditions particularly affected the phenolic compounds of fresh and lyophilized blueberry, but not the capsule, which may suggest that encapsulation protected the phenols present from alkaline conditions. Further studies on in vitro absorption of the remaining components in the intestine and their possible effects on oxidative stress biomarkers in in vivo models are suggested(AU)


Subject(s)
Tannins , Flavonoids , Blueberry Plants , Phenolic Compounds , Gastrointestinal Absorption , In Vitro Techniques , Chronic Disease , Digestion , Freeze Drying
11.
Bol. latinoam. Caribe plantas med. aromát ; 21(1): 41-50, ene. 2022. ilus, tab
Article in English | LILACS | ID: biblio-1370333

ABSTRACT

Solanum nudum Dunal (Solanaceae) is most commonly known andused by the population of the colombian Pacific coast as an antimalarial treatment. This article study into optimization and quantitative analysis of compounds steroidal over time of development of this species when grown in vitro and wild. A new steroidal compound named SN6 was elucidated by NMR and a new method of quantification of seven steroidal compounds (Diosgenone DONA and six steroids SNs) using HPLC-DAD-MS in extracts of cultures in vitroand wild was investigated. Biology activity of extracts was found to a range of antiplasmodial activity in FCB2 and NF-54 with inhibitory concentration (IC50) between (17.04 -100µg/mL) and cytotoxicity in U-937 of CC50 (7.18 -104.7µg/mL). This method creates the basis for the detection of seven sterols antiplasmodial present in extracts from S. nudum plant as a quality parameter in the control and expression of phytochemicals.


Solanum nudum Dunal (Solanaceae) es el más conocido y utilizado por la población de la costa del Pacífico colombiano como tratamiento antipalúdico. Este artículo estudia la optimización y el análisis cuantitativo de compuestos esteroides a lo largo del tiempo de desarrollo de esta especie cuando se cultiva in vitro y en forma silvestre. Un nuevo compuesto esteroideo llamado SN6 fue dilucidado por RMN y se investigó un nuevo método de cuantificación de siete compuestos esteroides (Diosgenone DONA y seis esteroides SN) usando HPLC-DAD-MS en extractos de cultivos in vitro y silvestres. La actividad biológica de los extractos se encontró en un rango de actividad antiplasmodial en FCB2 y NF-54 con concentración inhibitoria (IC50) entre (17.04 -100 µg/mL) y citotoxicidad en U-937 de CC50 (7.18 -104.7 µg/mL). Este método crea la base para la detección de siete esteroles antiplasmodiales presentes en extractos de planta de S. nudum como parámetro de calidad en el control y expresión de fitoquímicos.


Subject(s)
Steroids/analysis , Solanum/chemistry , Antimalarials/chemistry , In Vitro Techniques , Chromatography, High Pressure Liquid/methods , Solanum/growth & development , Tandem Mass Spectrometry , Phytochemicals , Antimalarials/pharmacology
12.
Acta sci., Health sci ; 44: e56960, Jan. 14, 2022.
Article in English | LILACS | ID: biblio-1367539

ABSTRACT

Colorectal cancer is the 4thcause of cancer death; with considering the growth process of this cancer and the necessity of early diagnosis, the purpose of the research is to state the LncRNA 00970, LncRNA UCAI,and the Wntgene before and after the treatment by 5-Azacytidine epigenetic medicine, to reach the biomarker in the very first steps of colorectal cancer. In this experiment, the human colon cancer cell line (HT29) treated with different concentrations of 5-aza-2'-deoxycytidine (5-aza-dC) was utilized to induce DNA demethylation; Quantitative PCR (qPCR) was used to measure LncRNA UCA1and LncRNA LINC00970 and Wntexpression. There was a significant relationship between the expression of LncRNA 00970, LncRNA UCAI,and the Wntgene and its effects on colorectal (p < 0.05). The Wntgene was treated by 1 and 10 of 5-Azacytidine epigenetic medicine, which then experienced decreases. In LncRNA UCAI and LncRNA00970 in dose 1 micromolar of 5-Azacytidine had decrement and increment of expressionrespectively that explains their efficiency but in treatment by dose 10 mM of this medicine, no significant LncRNA expression difference was detected, 5-azacitidine has a direct impact on its target genes and LncRNAs.Therefore, it can be used in the early diagnosis of colorectal cancer.


Subject(s)
In Vitro Techniques/methods , DNA/analysis , Colorectal Neoplasms/drug therapy , Colorectal Neoplasms/therapy , Colonic Neoplasms/diagnosis , Early Diagnosis , Azacitidine/analysis , Azacitidine/antagonists & inhibitors , Biomarkers , Colorectal Neoplasms/mortality , Cell Line/drug effects , Colonic Neoplasms/drug therapy , Colonic Neoplasms/therapy , Epigenomics , RNA, Long Noncoding , RNA, Long Noncoding/drug effects , Genes
13.
Braz. J. Pharm. Sci. (Online) ; 58: e191120, 2022. tab, graf
Article in English | LILACS | ID: biblio-1394048

ABSTRACT

Abstract The aim of the current study was to assess the physicochemical characteristics and wound healing activity of chitosan-polyvinyl alcohol (PVA) crosslinked hydrogel containing recombinant human epidermal growth factor (rh-EGF) or recombinant mouse epidermal growth factor (rm-EGF). The hydrogels were prepared and analyses were made of the morphological properties, viscosity, water absorption capacity, mechanical and bio-adhesive properties. The viscosity of the formulations varied between 14.400 - 48.500 cPs, with the greatest viscosity values determined in K2 formulation. F2 formulation showed the highest water absorption capacity. According to the studies of the mechanical properties, H2 formulation (0.153±0.018 N.mm) showed the greatest adhesiveness and E2 (0.245±0.001 mj/cm2) formulation, the highest bio-adhesion values. Hydrogels were cytocompatible considering in vitro cell viability values of over 76% on human keratinocyte cells (HaCaT, CVCL-0038) and of over 84% on human fibroblast cells (NIH 3T3, CRL-1658) used as a model cell line. According to the BrdU cell proliferation results, B1 (197.82±2.48%) formulation showed the greatest NIH 3T3 and C1 (167.43±5.89%) formulation exhibited the highest HaCaT cell proliferation ability. In addition, the scratch closure assay was performed to assess the wound healing efficiency of formulation and the results obtained in the study showed that F2 formulation including PEGylated rh-EGF had a highly effective role.


Subject(s)
Wound Healing , Hydrogels/analysis , Chitosan/chemical synthesis , Epidermal Growth Factor , Polyvinyl Alcohol/pharmacology , Wounds and Injuries/classification , In Vitro Techniques/methods , Cell Culture Techniques/methods , Cell Proliferation/genetics , Absorption
14.
São Paulo; s.n; s.n; 2022. 157 p. tab, graf, ilus.
Thesis in English | LILACS | ID: biblio-1380998

ABSTRACT

Melanoma accounts for 3% of skin neoplasms and is the leading cause of death from skin disorders worldwide. The high mortality rate associated with this disease stems from the high capacity of melanoma patients to develop metastases and treatment relapse with inhibitors of the MAPK signaling pathway (such as BRAF inhibitors), commonly used in melanoma therapy. Thus, the investigation of genes involved in the mechanisms of melanoma development is essential for new and more effective therapeutic strategies. Hence, we describe in this thesis two projects involving the genes SIN3B and IRF4 as possible biomarkers for cutaneous melanoma. Initially, through bioinformatics analyses performed by our group, an upregulation of SIN3B was found in metastatic melanomas. This result together with the understanding of SIN3B role in regulating gene expression and oncogenic transformation, prompted us to describe in this thesis some mechanisms by which SIN3B may influence melanoma development. We then sought to characterize the gene function using SIN3B-deleted cells, generated by the CRISPR-Cas9 methodology. Initially, we observed increased SIN3B expression in BRAF-mutant metastatic melanomas, where we noted that the long splicing variant of the gene (NM_001297595.1) was effectively prevalent in melanomas. Subsequently, we designed gRNAs between the exons 2 and 3 of the human SIN3B gene and engineered three knockout clones and three control clones (containing empty lentiCRISPRv2 plasmid) from different melanoma cell lines (SKMEL28, A2058, and A375). Through functional analyses, it was observed that the absence of the gene did not interfere in the proliferation of tumor cells; however, it led to a decrease in invasive properties. These results were verified by Boyden chamber assays and transcriptome analysis (total RNA sequencing of deleted cells), where a decrease in migration and motility pathways was observed. Additionally, a screening of synthetically lethal genes with SIN3B was performed with a genome wide CRISPR library. These results showed that USP7 and STK11 genes, which belong to the FoxO signaling pathway, were essential in SIN3B-depleted melanoma cells. Finally, through a collaborative project with the Wellcome Trust Sanger Institute, previous large-scale sequencing analyses demonstrated that deletion of the IRF4 gene was lethal for melanoma cells. Accordingly, we performed IRF4 silencing in vitro and noticed that the lack of IRF4 promotes cell death and apoptosis, independently of MYC and MITF, known in the literature to be downstream targets of this gene. Therefore, these data suggest that IRF4 plays a vital role in melanoma cell survival. Taken together, both works herein described in this thesis demonstrate how CRISPR-Cas9 can be applied to study the functions and mechanisms of genes involved in melanoma progression, collectively helping in the development of more effective therapeutic strategies for this tumor


O melanoma representa 3% dos tipos de neoplasias cutâneas e é a maior causa das mortes por distúrbios de pele no mundo. A alta taxa de mortalidade associada à essa doença advém da alta capacidade de pacientes com melanoma desenvolverem metástases, e apresentarem recidiva após tratamento com inibidores da via de sinalização MAPK (como da proteína BRAF), comumente utilizados no tratamento de pacientes metastáticos. Assim, a investigação de genes envolvidos nos mecanismos de desenvolvimento do melanoma é primordial para novas estratégias terapêuticas mais efetivas. Dessa forma, descrevemos no presente trabalho dois projetos envolvendo os genes SIN3B e IRF4 como possíveis biomarcadores para melanoma cutâneo. Em análises prévias de bioinformática realizados pelo nosso grupo, SIN3B foi identificado tendo maior expressão em melanomas metastáticos. Além disso, diversos estudos mostraram que o gene está envolvido na regulação da expressão gênica e transformação oncogênica. Dessa forma, descrevemos nessa tese alguns mecanismos pelos quais SIN3B pode influenciar no desenvolvimento do melanoma, através da caracterização funcional de células SIN3B-deletadas pela metodologia CRISPR-Cas9. Inicialmente, observamos aumento na expressão de SIN3B em melanomas metastáticos BRAF-mutados, onde notamos que a variante de splicing longa do gene (NM_001297595.1), era efetivamente prevalente em melanomas. Assim, desenhamos sequências de RNA guias entre os éxons 2 e 3 do gene SIN3B humano e, obtivemos três clones knockout e outros três clones controle (contendo plasmídeo vazio) em diferentes linhagens de melanoma (SKMEL28, A2058 e A375), para caracterização funcional. Observou-se que a ausência do gene não interferiu na proliferação das células tumorais, contudo, acarretou na diminuição de processos invasivos. Esses resultados foram averiguados através de ensaios em câmara de Boyden e análises de transcriptoma (sequenciamento de RNA total das células deletadas), onde notou-se diminuição das vias de migração e motilidade. Adicionalmente, um rastreamento de genes sinteticamente letais com SIN3B foi realizado com uma biblioteca de CRISPR capaz de silenciar todo o genoma. Esses resultados mostraram que os genes USP7 e STK11, ambos pertencentes à via de sinalização de FoxO, são essenciais nas células SIN3B deletadas. Por fim, através de um projeto colaborativo com o Wellcome Trust Sanger Institute, análises prévias de sequenciamento de larga escala demonstraram que a deleção do gene IRF4 era letal para células de melanoma. Dessa forma, realizamos o silenciamento de IRF4 in vitro e notamos que a ausência do gene promove morte celular e apoptose, independentemente de MYC e MITF, conhecidos na literatura por serem alvos downstream do gene. Portanto, esses dados sugerem que IRF4 tem um papel importante na sobrevivência de células de melanoma. Em conjunto, ambos trabalhos descritos nessa tese, demonstram como a metodologia CRISPR-Cas9 pode auxiliar no entendimento de processos importantes para a malignidade do melanoma e contribuir para estratégias terapêuticas mais efetivas para esse tumor


Subject(s)
Skin Neoplasms/complications , Methodology as a Subject , Melanoma/pathology , Neoplasm Metastasis , Neoplasms , Patients/classification , Skin , In Vitro Techniques/methods , Biomarkers/analysis , Gene Expression , Cell Survival , Sequence Analysis, RNA/instrumentation , Computational Biology/methods , Absenteeism , Clustered Regularly Interspaced Short Palindromic Repeats
15.
Braz. J. Pharm. Sci. (Online) ; 58: e191024, 2022. tab, graf
Article in English | LILACS | ID: biblio-1394036

ABSTRACT

Abstract Posaconazole exerts an extended spectrum of antifungal activity against various strains of clinically relevant moulds and yeasts. In recent years, antifungal triazole posaconazole has become increasingly important for the prophylaxis and treatment of systemic mycoses. After oral administration of posaconazole, absolute bioavailability has been estimated to range from 8% to 47%. Pharmaceutical co-crystallization is a promising approach for improving dissolution rate or manipulating other physical properties of API. The objective of this study is to improve the dissolution rate of posaconazole by co-crystallization. A 1:1 stoichiometric co-crystals of adipic acid were prepared by solvent assisted grinding method. The prepared co-crystals were subjected to solid-state characterization by FTIR, PXRD and DSC studies. The physicochemical properties of posaconazole and co-crystals were assessed in terms of melting point, flowability and dissolution rate. The results indicated improvement in flow property and dissolution rate. In vitro dissolution profile of co-crystals showed a significant increased dissolution of posaconazole from initial period in 0.1 N hydrochloric acid solution. The dissolution efficiency for posaconazole-adipic acid co-crystal was 61.65 % against posaconazole, 46.58 %. Thus, co-crystallization can be a promising approach to prepare posaconazole-adipic acid co-crystals with improved physicochemical properties.


Subject(s)
Administration, Oral , Crystallization/instrumentation , Hydrochloric Acid , Sprains and Strains/diagnosis , Yeasts/classification , In Vitro Techniques/methods , Pharmaceutical Preparations , Biological Availability , Spectroscopy, Fourier Transform Infrared , Efficiency , Dissolution , Mycoses/pathology
16.
Braz. J. Pharm. Sci. (Online) ; 58: e19517, 2022. tab, graf
Article in English | LILACS | ID: biblio-1383995

ABSTRACT

Nordihydroguaiaretic acid (NDGA) is a natural product obtained by the alkaline extraction of dried plants of Larrea tridentata species. Due to the biological properties presented, such as antioxidant, anti-inflammatory, antiviral and cytotoxic capacity, this compound is being increasingly studied. In this review, it was evaluated the benefits of NDGA against different animal models. Besides that, it was found that this compound has antitumor activity similar to its synthetic derivative terameprocol in prostate tumors. The hypoglycemic effect may be evidenced by the inhibition of sugar uptake by NDGA; in obesity, studies have observed that NDGA presented a positive regulatory effect for Peroxisome proliferator-activated receptors (PPAR-α) involved in the oxidation of hepatic fatty acids and reduced the expression of lipogenic genes. Regarding its antioxidant potential, its mechanism is related to the ability to in vitro scavenging reactive substances. Although there are several studies demonstrating the benefits of using NDGA, there are also reports of its toxicity, mainly of liver damage and nephrotoxicity


Subject(s)
Masoprocol/analysis , Chemical Phenomena , Antiviral Agents/pharmacology , Plants/classification , Biological Products/analysis , In Vitro Techniques/methods , Models, Animal , Toxicity , Hypoglycemic Agents/pharmacology , Neoplasms , Antioxidants/pharmacology
17.
Pesqui. bras. odontopediatria clín. integr ; 22: e210112, 2022. tab, graf
Article in English | LILACS, BBO | ID: biblio-1386810

ABSTRACT

Abstract Objective: To evaluate the efficacy of silver diamine fluoride (SDF) in arresting dentin caries lesions when applied under different concentrations and times. Material and Methods: Forty-two bovine blocks were selected and fixed in 24-well plates. Each well received a mixed bacterial inoculum added to the culture medium with 5% sucrose. The plates were incubated in microaerophilia (7 days) for caries formation, confirmed by micro-CT (M1). SDF was applied over the carious lesions for different times and concentrations (n=6): SDF 30% - immediate removal, 1 minute and 3 minutes; SDF 38%, - immediate removal, 1 minute and 3 minutes. The group without treatment was the control. Then, the samples were again scanned by micro-CT (M2) and submitted to a second cariogenic challenge for 21 days. Then, a final scan was performed (M3). Results: Mean pH at the culture medium and lesion depth were compared using Kruskal-Wallis and Wilcoxon tests. 38% SDF showed the lowest metabolic activity of the biofilm. All 38% groups and 30% 1 and 3 minutes did not show an increase in mean lesion depth comparing M3 with M1. However, only 30% 3 minutes and 38% 1 and 3 minutes showed a significant reduction of lesion depth. Conclusion: The minimum application time of 30% SDF to arrest dentin caries lesion was 1 minute, while 38% SDF arrested with application and immediate removal.


Subject(s)
Animals , Cattle , Tooth Remineralization , Cariostatic Agents/therapeutic use , Dental Caries/epidemiology , Dentin , Diamines/chemistry , Fluorides/chemistry , Silver/therapeutic use , In Vitro Techniques/methods , Longitudinal Studies , Statistics, Nonparametric , Biofilms , X-Ray Microtomography/instrumentation
18.
Braz. J. Pharm. Sci. (Online) ; 58: e19898, 2022. tab, graf
Article in English | LILACS | ID: biblio-1394061

ABSTRACT

Abstract A study was carried out to synthesize silver nanoparticles (Ag-NPs) using plant extracts and to explore their pharmaceutical application as antibacterial agents. Dried leaves of Mentha arvensis Linn. were powdered and extracts were prepared using three different organic solvents. Preliminary screening on antibacterial activity by agar well diffusion method indicated that the methanolic extract possessed higher potential than the other two. Phytochemical analysis of the leaf extract revealed that it possesses tannins, steroids, terpenoids and flavonoids. Ag-NPs prepared using this extract were of spherical to cuboidal shape ranging in size from 40 to 70 nm. It retained essential chemical groups and had the required surface plasmon resonance. Further to the antimicrobial assay, the Ag-NPs, in contrast to the whole methanolic leaf extract, showed better (100% vs. 80%) bactericidal activity against the indicator organisms. The zones of growth inhibition for gram positive bacteria ranged between 14±0.6 and 23±0.3, while it was between 12±0.6 and 22±0.2 for gram negative bacteria. The Ag-NPs presented the minimum inhibitory concentration values of 250 µg/mL for gram positive bacteria and 250 - 500 µg/mL for gram negative bacteria. These findings suggest that the natural compounds present in M. arvensis L. possess the potency to facilitate the synthesis and antibacterial action of Ag-NPs.


Subject(s)
Silver , Plant Extracts/adverse effects , Mentha/adverse effects , Nanoparticles/analysis , Anti-Bacterial Agents/pharmacology , In Vitro Techniques/instrumentation , Pharmaceutical Preparations/analysis , Phytochemicals/administration & dosage , Gram-Negative Bacteria/classification , Gram-Positive Bacteria/classification
19.
Braz. J. Pharm. Sci. (Online) ; 58: e19801, 2022. tab, graf
Article in English | LILACS | ID: biblio-1394060

ABSTRACT

Abstract In the recent past, drug delivery through nanoparticles is considered an effective tool to treat various diseases. Biopolymeric nanoparticles such as protein based nanoparticles have vital role as drug carrier as it is non-antigenic, and easily biodegradable. Curcumin, plant polyphenolic anticancerous compound was loaded into the casein nanoparticles by coacervation method. Particle size and surface charge of spherical casein nanoparticles as observed to be 201.4 nm and -86.9 mV. The loading efficiency of curcumin loaded casein nanoparticles was found to 85.05 %. In vitro drug release was performed at different pH (7.4 and 3.0), and the cumulative release was observed to be 24.8 and 20.13% and at different temperatures (25°C and 37°C), the cumulative release was observed to be 24.8 and 28.60 % respectively in 48 h. Curcumin release from casein nanoparticles was shown to be in a steady, and prolonged rate. The nanoparticles were observed to have an effective antimocrobial activity than curcumin in free form. The drug loaded casein nanoparticles were found to be potent particles to protect cells from hydrogen peroxide and UV light damage. The cytotoxic activity of nanoparticles on MCF7 and A549 cells were assayed and was observed to have an IC50 value of 609 and 825.2µg/ml. Cell death was observed to be through apoptosis, accompanied by DNA fragmentation.


Subject(s)
In Vitro Techniques/methods , Caseins/agonists , Curcumin/classification , Nanoparticles/classification , Ultraviolet Rays/adverse effects , Apoptosis , Inhibitory Concentration 50 , Drug Liberation , A549 Cells
20.
Braz. J. Pharm. Sci. (Online) ; 58: e191009, 2022. tab, graf
Article in English | LILACS | ID: biblio-1394059

ABSTRACT

Nizatidine is an anti-secretogogue and a gastroprotective drug with a half-life of 1-2 h and is well absorbed in the stomach. This study aimed to optimize the process and develop floating microparticles of nizatidine that are based on low methoxyl pectin. Oil-in-oil dispersion method and Taguchi orthogonal array design were employed, and the prolonged residence time of the microparticles in the stomach was demonstrated. The constraints for independent variables, viz. A-polymer, B-internal solvent volume, C-surfactant, D-stirring rate and E-stirring time were set to generate the experimental runs. Particle size, percentage yield, micromeritic properties, entrapment efficiency, in vitro buoyancy and in vitro release were characterized. Surface morphology, zeta potential, in vitro release kinetics and in vivo floating performance of the optimized formulation was examined. The microparticles were free-flowing, irregular in shape and had a mean particle size distribution of 73-187 µ. Low methoxyl pectin played a predominant role in achieving buoyancy and optimum gastric retention for the modified release of the drug, suggesting Korsmeyer-Peppas model as the possible release mechanism. In vivo radiographic study in rabbits revealed that the drug was retained in the stomach for a period of 6 h. These results indicate that nizatidine floating microparticulate system provides modified drug release for the effective treatment of gastric ulcer


Subject(s)
Animals , Male , Female , Rabbits , Stomach/drug effects , Nizatidine/antagonists & inhibitors , Efficiency/classification , Solvents/adverse effects , Stomach Ulcer/pathology , In Vitro Techniques/instrumentation , Pharmaceutical Preparations/administration & dosage , Kinetics , Spectroscopy, Fourier Transform Infrared/methods , Drug Liberation
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