RESUMEN
Background: Prodigiosin has been demonstrated to be an important candidate in investigating anticancer drugs and in many other applications in recent years. However, industrial production of prodigiosin has not been achieved. In this study, we found a prodigiosin-producing strain, Serratia marcescens FZSF02, and its fermentation strategies were studied to achieve the maximum yield of prodigiosin. Results: When the culture medium consisted of 16.97 g/L of peanut powder, 16.02 g/L of beef extract, and 11.29 mL/L of olive oil, prodigiosin reached a yield of 13.622 ± 236 mg/L after culturing at 26 °C for 72 h. Furthermore, when 10 mL/L olive oil was added to the fermentation broth at the 24th hour of fermentation, the maximum prodigiosin production of 15,420.9 mg/L was obtained, which was 9.3-fold higher than the initial level before medium optimization. More than 60% of the prodigiosin produced with this optimized fermentation strategy was in the form of pigment pellets. To the best of our knowledge, this is the first report on this phenomenon of pigment pellet formation, which made it much easier to extract prodigiosin at low cost. Prodigiosin was then purified and identified by absorption spectroscopy, HPLC, and LCMS. Purified prodigiosin obtained in this study showed anticancer activity in separate experiments on several human cell cultures: A549, K562, HL60, HepG2, and HCT116. Conclusions: This is a promising strain for producing prodigiosin. The prodigiosin has potential in anticancer medicine studies.
Asunto(s)
Prodigiosina/biosíntesis , Prodigiosina/farmacología , Serratia marcescens/metabolismo , Antineoplásicos/farmacología , Arachis/química , Polvos , Prodigiosina/aislamiento & purificación , Espectrometría de Masas , Células Tumorales Cultivadas/efectos de los fármacos , Cromatografía Líquida de Alta Presión , Cromatografía Liquida , Técnicas de Cultivo de Célula , Fermentación , Aceite de Oliva/química , Acetatos , NitrógenoRESUMEN
BACKGROUND: Ovarian cancer is the most lethal of all gynecologic malignancies. The relationship between sexual steroids receptors and ovarian cancer progression has been largely evaluated. The presence of progesterone receptors has been associated with an increase of a disease-free period and overall survival in patients with ovarian carcinoma. In the present study, primary cultures of ovarian carcinoma obtained from 35 patients diagnosed with epithelial ovarian cancer were evaluated for cell survival after treatment with 10- 8 M of 17ß-estradiol, progesterone, testosterone and dihydrotestosterone. RESULTS: The results were analyzed considering histological subtypes: low grade serous, high grade serous, endometrioid and mucinous carcinoma; clear cell carcinoma was not included due to failure in obtaining successful cultures of this subtype. A significant reduction of cell survival was observed after progesterone treatment in endometrioid ovarian carcinoma. Changes were not observed in low grade serous, high grade serous and mucinous carcinoma. The effect of progesterone was related to the presence of progesterone receptor (PR), a 43% reduction in the cell number was observed in PR (+) endometrioid ovarian carcinoma. CONCLUSIONS: This study supports the importance of progesterone and the presence of progesterone receptor in the reduction of ovarian cancer progression in the endometrioid ovarian carcinoma.
Asunto(s)
Antineoplásicos Hormonales/farmacología , Carcinoma Endometrioide/patología , Supervivencia Celular/efectos de los fármacos , Neoplasias Ováricas/patología , Progesterona/farmacología , Adulto , Carcinoma Endometrioide/metabolismo , Carcinoma Epitelial de Ovario/metabolismo , Carcinoma Epitelial de Ovario/patología , Proliferación Celular/efectos de los fármacos , Femenino , Humanos , Persona de Mediana Edad , Neoplasias Ováricas/metabolismo , Estudios Prospectivos , Receptores de Progesterona/metabolismo , Receptores de Esteroides/metabolismo , Células Tumorales Cultivadas/efectos de los fármacosRESUMEN
The Bacille Calmette-Guérin (BCG) vaccine comprises a family of genetically different strains derived by the loss of genomic regions (RDs) and other mutations. In BCG Moreau, loss of RD16 inactivates rv3405c * , encoding a transcriptional repressor that negatively regulates the expression of Rv3406, an alkyl sulfatase. To evaluate the impact of this loss on the BCG and host cell viability and the cytokine profile, THP-1 cells were infected with BCG Moreau (harbouring the empty vector) and a complemented strain carrying a functional copy of rv3405c. Viability of the host cells and bacteria as well as the pattern of cytokine secretion were evaluated. Our results show that the viability of BCG Moreau is higher than that of the complemented strain in an axenic medium, suggesting a possible functional gain associated with the constitutive expression of Rv3406. Viability of the host cells did not vary significantly between recombinant strains, but differences in the profiles of the cytokine secretion (IL-1ß and IL-6) were observed. Our results suggest an example of a functional gain due to gene loss contributing to the elucidation of the impact of RD16 on the physiology of BCG Moreau.
Asunto(s)
Vacuna BCG/farmacología , Supervivencia Celular/genética , Citocinas/efectos de los fármacos , Mutación con Ganancia de Función/genética , Macrófagos/microbiología , Mycobacterium bovis/genética , Transcripción Genética/genética , Vacuna BCG/genética , Supervivencia Celular/efectos de los fármacos , Citocinas/genética , Mutación con Ganancia de Función/efectos de los fármacos , Humanos , Mycobacterium bovis/fisiología , Factores de Tiempo , Transcripción Genética/efectos de los fármacos , Células Tumorales Cultivadas/efectos de los fármacos , Células Tumorales Cultivadas/microbiologíaRESUMEN
Introdução: Bisfosfonatos são fármacos utilizados para o tratamento de enfermidades ósseas, como a osteoporose e metástases ósseas, em razão do seu mecanismo de ação, que consiste na diminuição do processo de reabsorção do osso. Outros estudos verificaram que bisfosfonatos de alta potência, como o zoledronato, poderiam auxiliar no tratamento de outras enfermidades malignas por causa da promoção de um efeito antiproliferativo. Objetivo: Este estudo in vitro objetivou avaliar a atividade antiproliferativa de zoledronato em diferentes linhagens de células tumorais. Método: Nove linhagens humanas (U251; MCF7; NCI/ADR-RES; 786-0; NCI-H460; PC-3; OVCAR-3; HT29; K-562 e HaCaT) foram submetidas ao tratamento com as concentrações de 0,12; 1,2; 12 e 120 µM de zoledronato e tiveram sua atividade proliferativa avaliada após 48 horas, utilizando-se o corante sulforrodamina B. Resultados: Verificou-se que as concentrações de 12 µM e 120 µM de zoledronato foram eficazes para a redução em 50% e 100%, respectivamente, da proliferação das células 786-0 (carcinoma renal). A maior concentração de zoledronato (120 µM) promoveu um efeito citostático (redução da proliferação celular em 50%) para as células HaCaT (queratinócito humano não tumoral), HT-29 (carcinoma de cólon), NCI-ADR/ RES (adenocarcinoma de ovário com fenótipo de multirresistência) e NCI-H460 (carcinoma pulmonar). Conclusão: Esses resultados sugerem um promissor efeito auxiliar do zoledronato para o tratamento de alguns tipos de tumores; estudos complementares in vitro e in vivo são necessários para a validação dessa hipótese.
Introduction: Bisphosphonates are used in the treatment of bone diseases such as osteoporosis and bone metastases, because of their ability to inhibit bone resorption. There is evidence that high-potency bisphosphonates, such as zoledronate, are useful in the treatment of other malignancies because they have an antiproliferative effect. Objective:To evaluate the antiproliferative activity of zoledronate in different tumor cell lines. Method: This was an in vitro study in which nine human cell lines (U251, MCF7, NCI/ ADR-RES, 786-0, NCI-H460, PC-3, OVCAR-3, HT29, K-562, and HaCaT) were treated with of 0.12, 1.2, 12, and 120 µM of zoledronate, their proliferative activity being evaluated 48 h later with sulforhodamine B assay. Results: At the 12 µM and 120 µM doses, zoledronate effectively reduced the proliferation of 786-0 (renal carcinoma) cells by 50% and 100%, respectively. At the highest concentration (120 µM), zoledronate had a cytostatic effect (50% reduction in cell proliferation) on HaCaT (non-tumor human keratinocyte), HT-29 (colon carcinoma), NCI-ADR/ RES (multidrug-resistant ovarian adenocarcinoma), and NCI-H460 (lung carcinoma) cells. Conclusion: These results suggest a promising auxiliary effect of zoledronate for the treatment of some tumors. Further in vitro and in vivo studies are needed in order to test that hypothesis.
Introducción: Los bisfosfonatos son fármacos utilizados para el tratamiento de enfermedades óseas, como la osteoporosis y metástasis óseas debido a su mecanismo de acción, que consiste en la disminución del proceso de reabsorción del hueso. Otros estudios observaron que los bisfosfonatos de alta potencia, como el zoledronato, podrían ayudar en el tratamiento de otras enfermedades malignas debido a la promoción de un efecto antiproliferativo. Objetivo: Este estudio in vitro objetivó evaluar la actividad antiproliferativa de zoledronato en diferentes linajes de células tumorales. Método: Los nueve humano linajes (U251, MCF7, NCI / ADR-RES, 786-0, NCI-H460, PC-3, OVCAR-3, HT29, K-562 and HaCaT) se sometieron al tratamiento con las concentraciones de 0,12; 1,2; 12 y 120 µM de zoledronato y tuvieron su actividad proliferativa evaluada después de 48 horas utilizando el colorante sulforrodamina B. Resultados: Se comprobó que las concentraciones de 12 µM y 120 µM de zoledronato fueron efectivas para reducir en un 50% y un 100%, respectivamente, de la proliferación de las células 786-0 (carcinoma renal). La mayor concentración de zoledronato (120 µM) promovió un efecto citostático (reducción de la proliferación celular en un 50%) para las células HaCaT (queratinocito humano no tumoral), HT-29 (carcinoma de colon), NCI-ADR/RES (adenocarcinoma de ovário con fenótipo de multirresistencia) y NCI-H460 (carcinoma pulmonar). Conclusión: Estos resultados sugieren un prometedor efecto auxiliar del zoledronato para el tratamiento de algunos tumores; se requieren más estudios in vitro e in vivo para validar esta hipótesis
Asunto(s)
Humanos , Proliferación Celular/efectos de los fármacos , Difosfonatos , Técnicas In Vitro , Células Tumorales Cultivadas/efectos de los fármacos , Línea Celular Tumoral/efectos de los fármacosRESUMEN
The Bacille Calmette-Guérin (BCG) vaccine comprises a family of genetically different strains derived by the loss of genomic regions (RDs) and other mutations. In BCG Moreau, loss of RD16 inactivates rv3405c * , encoding a transcriptional repressor that negatively regulates the expression of Rv3406, an alkyl sulfatase. To evaluate the impact of this loss on the BCG and host cell viability and the cytokine profile, THP-1 cells were infected with BCG Moreau (harbouring the empty vector) and a complemented strain carrying a functional copy of rv3405c. Viability of the host cells and bacteria as well as the pattern of cytokine secretion were evaluated. Our results show that the viability of BCG Moreau is higher than that of the complemented strain in an axenic medium, suggesting a possible functional gain associated with the constitutive expression of Rv3406. Viability of the host cells did not vary significantly between recombinant strains, but differences in the profiles of the cytokine secretion (IL-1β and IL-6) were observed. Our results suggest an example of a functional gain due to gene loss contributing to the elucidation of the impact of RD16 on the physiology of BCG Moreau.
Asunto(s)
Humanos , Transcripción Genética/genética , Vacuna BCG/farmacología , Supervivencia Celular/genética , Citocinas/efectos de los fármacos , Mutación con Ganancia de Función/genética , Macrófagos/microbiología , Mycobacterium bovis/genética , Factores de Tiempo , Transcripción Genética/efectos de los fármacos , Células Tumorales Cultivadas/efectos de los fármacos , Células Tumorales Cultivadas/microbiología , Vacuna BCG/genética , Supervivencia Celular/efectos de los fármacos , Citocinas/genética , Mutación con Ganancia de Función/efectos de los fármacos , Mycobacterium bovis/fisiologíaRESUMEN
Photodynamic therapy (PDT), a promising treatment option for cancer, involves the activation of a photosensitizer (PS) by local irradiation with visible light. Excitation of the PS leads to a series of photochemical reactions and consequently the local generation of harmful reactive oxygen species (ROS) causing limited or none systemic defects. However, the development of resistance to this promising therapy has slowed down its translation into the clinical practice. Thus, there is an increase need in understanding of the molecular mechanism underlying resistance to PDT. Here, we aimed to examine whether a relationship exists between PDT outcome and ROS-involvement in the resistance mechanism in photosensitized cancer cells. In order to recapitulate tumor architecture of the respective original tumor, we developed a multicellular three-dimensional spheroid system comprising a normoxic periphery, surrounding a hypoxic core. Using Me-ALA, a prodrug of the PS PpIX, in human colorectal spheroids we demonstrate that HIF-1 transcriptional activity was strongly up-regulated and mediates PDT resistant phenotype. RNAi knockdown of HIF-1 impairs resistance to PDT. Oxidative stress-mediated activation of ERK1/2 followed PDT was involved on positive modulation of HIF-1 transcriptional activity after photodynamic treatment. ROS scavenging and MEK/ERK pathway inhibition abrogated the PDT-mediated HIF-1 upregulation. Together our data demonstrate that resistance to PDT is in part mediated by the activation of a ROS-ERK1/2-HIF-1 axis, thus, identifying novel therapeutic targets that could be used in combination with PDT.
Asunto(s)
Neoplasias Colorrectales/genética , Resistencia a Antineoplásicos , Factor 1 Inducible por Hipoxia/genética , Fotoquimioterapia/métodos , Especies Reactivas de Oxígeno/metabolismo , Células Tumorales Cultivadas/citología , Técnicas de Cultivo de Célula , Línea Celular Tumoral , Proliferación Celular , Supervivencia Celular , Neoplasias Colorrectales/tratamiento farmacológico , Neoplasias Colorrectales/metabolismo , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Humanos , Factor 1 Inducible por Hipoxia/metabolismo , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , Modelos Biológicos , Esferoides Celulares , Células Tumorales Cultivadas/efectos de los fármacos , Regulación hacia ArribaRESUMEN
Currently, mammary neoplasms in female canines are a serious problem in veterinary clinics. In addition, the canine species is an excellent disease model for human oncology because of the biological and genetic similarities between the species. Cytogenetics has allowed further study of the characterization of neoplasms in canines. We hypothesized that the use of a direct preparation protocol for mitotic chromosome analysis would provide a simple and low cost protocol for use in all laboratories. The objective of this method is to display in a few hours of dividing cells just like the time of collection since cell division in tissue can be obtained. Ten female canines with the spontaneous occurrence of mammary neoplasia were used to test a pioneering direct preparation protocol to obtain mitotic chromosomes. The excised breast tumor tissue fragments were subjected to the protocol consisting of treatment with colchicine, treatment with hypotonic solution, and fixation. Mitotic chromosomes were absent in cell suspensions of only two samples among the 10 materials analyzed, based on the analysis of five blades for each preparation obtained. So, the cell suspension obtained allowed for the observation of eight tissue samples viable for cytogenetic analysis, five of which had excellent numbers of mitotic chromosomes. However, the technique was unsuccessful in producing high-quality cell suspensions because of inadequate condensation and scattering of chromosomes. While adjustments to methodological procedures are needed, this protocol represents a low cost and simplified method to study the cytogenetics of canine tumors.
Asunto(s)
Carcinoma Papilar/ultraestructura , Carcinosarcoma/ultraestructura , Cromosomas de los Mamíferos/ultraestructura , Análisis Citogenético/métodos , Neoplasias Mamarias Animales/ultraestructura , Metafase/efectos de los fármacos , Animales , Carcinoma Papilar/genética , Carcinoma Papilar/patología , Carcinosarcoma/genética , Carcinosarcoma/patología , Colchicina/farmacología , Perros , Femenino , Humanos , Soluciones Hipotónicas/farmacología , Glándulas Mamarias Animales/patología , Glándulas Mamarias Animales/ultraestructura , Neoplasias Mamarias Animales/genética , Neoplasias Mamarias Animales/patología , Células Tumorales Cultivadas/efectos de los fármacos , Células Tumorales Cultivadas/ultraestructuraRESUMEN
O artigo analisa o livro Boys in white: student culture in medical school, de Howard S. Becker, Blanche Geer, Everett C. Hughes e Anselm Strauss, considerado um dos modelos de pesquisa qualitativa em sociologia. A análise aborda as trajetórias dos autores, do livro, da pesquisa qualitativa e dos estudantes de medicina, enfatizando sua importância nas origens da sociologia médica e da sociologia da educação médica. Na trajetória dos autores são apresentados aspectos biobibliográficos; na da pesquisa qualitativa, o modo como essa metodologia de investigação atravessa a construção do trabalho de campo; e na dos estudantes, sua forma de atravessar os primeiros anos da escola médica e construir sua própria “cultura do estudante”.
This article analyzes Boys in white: student culture in medical schoolby Howard S. Becker, Blanche Geer, Everett C. Hughes and Anselm Strauss, considered a model of qualitative research in sociology. The analysis investigates the trajectories of the authors, the book, qualitative analysis, and the medical students, emphasizing their importance in the origins of medical sociology and the sociology of medical education. In the trajectory of the authors, bibliographical information is given. The trajectory of qualitative research focuses on how this methodology influences the construction of the field. The investigation of the students’ trajectory shows how they progress through their first years at medical school to build their own student culture.
Asunto(s)
Animales , Femenino , Ratones , Adenocarcinoma/tratamiento farmacológico , Antimetabolitos Antineoplásicos/farmacología , Antineoplásicos Hormonales/farmacología , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Apoptosis/efectos de los fármacos , Neoplasias de la Mama/tratamiento farmacológico , Estrógenos , Antagonistas de Estrógenos/farmacología , Inhibidores de Crecimiento/farmacología , Neoplasias Hormono-Dependientes/tratamiento farmacológico , Fenilacetatos/farmacología , /biosíntesis , Tamoxifeno/farmacología , Adenocarcinoma/patología , Antimetabolitos Antineoplásicos/administración & dosificación , Antineoplásicos Hormonales/administración & dosificación , Protocolos de Quimioterapia Combinada Antineoplásica/farmacología , Neoplasias de la Mama/patología , Diferenciación Celular/efectos de los fármacos , División Celular/efectos de los fármacos , Sinergismo Farmacológico , Genes ras , Ratones Desnudos , Trasplante de Neoplasias , Neoplasias Hormono-Dependientes/patología , /fisiología , Fenilacetatos/administración & dosificación , /genética , Transfección , Tamoxifeno/administración & dosificación , Células Tumorales Cultivadas/efectos de los fármacosRESUMEN
Se analiza el significado del concepto de “obsesión” en el alienismo del siglo XIX. Desde el punto de vista clínico, la descripción de Esquirol fue completada por otros autores (Jules Falret, Legrand du Saulle). En el ámbito de la reflexión psicopatológica, el alienismo francés, con el delirio emotivo de Morel o la psicastenia de Janet, defendió la teoría emocional, frente al trastorno intelectual propuesto por los médicos alemanes. Finalmente, se insiste en la importancia del marco cultural en la aparición de los síntomas obsesivos y de su interpretación. En este sentido, se estudian las relaciones de los escrúpulos religiosos con la melancolía o la aparición de categorías diagnósticas sometidas a los códigos y mentalidades fin-de-siècle.
The article analyses the significance of the concept of “obsession” in nineteenth-century alienism. From a clinical point of view, Esquirol’s description was completed by other authors (Jules Falret, Legrand du Saulle). In the area of psychopathological studies, French alienism, with Morel’s emotional delirium or Janet’s psychasthenia, defended the emotional theory, as opposed to the intellectual disorder proposed by German doctors. Lastly, the importance of the cultural framework is stressed in the appearance of obsessive symptoms and their interpretation. Along these lines, the article discusses the relationship of religious scruples to melancholy or the appearance of diagnostic categories subject to fin de siècle codes and mentalities.
Asunto(s)
Humanos , Antineoplásicos/farmacología , ADN Complementario/genética , Floxuridina/farmacología , Factor de Crecimiento Derivado de Plaquetas/genética , Timidina Fosforilasa/genética , Comunicación Celular , Ensayos de Selección de Medicamentos Antitumorales , Transfección , Células Tumorales Cultivadas/efectos de los fármacos , Células Tumorales Cultivadas/enzimología , Células Tumorales Cultivadas/patologíaRESUMEN
Objetivo. Analizar la percepción que el prestador de servicios de salud y el adulto mayor (AM) tienen sobre el maltrato al AM en los servicios públicos de salud, en ciudades seleccionadas de México. Material y métodos. De 2009 a 2012 se realizó un estudio con diseño cualitativo y estrategia de triangulación de fuentes de datos; se efectuaron entrevistas semiestructuradas a 13 prestadores y a 12 ancianos para recuperar su experiencia en el tema. El análisis utilizó procedimientos de la Teoría Fundamentada. Resultados. El maltrato contra el AM es una práctica naturalizada por el personal y por el anciano, la cual se manifiesta de formas diversas. Conclusiones. La institucionalización, profesionalización histórica y falta de conciencia sobre las necesidades de los AM demandan cambios de planeación, organización y supervisión del Sistema de Salud. El personal requiere intervenciones de formación, capacitación y cambio de actitudes/comportamiento, para otorgar atención integral, digna, humana y de respeto a los Derechos Humanos de los AM.
Objective. To analyze the health care providers (HCP) and elderly patients' perceptions about abuse of the elderly by health personnel of public health services, in selected cities in Mexico. Materials and methods. A qualitative study and a strategy of data triangulation were performed during 2009 and 2012; 13 HCPs and 12 elders were interviewed, in order to obtain their experience regarding elder abuse. Grounded Theory proceedings were used for the analysis. Results. Elder abuse is a naturalized practice, from HCP and elderly people's point of view; these perceptions are showed in different ways. Conclusion. Institutionalization, historical professionalization and lack of consciousness about needs of the elderly (sociocultural and economic), require changes in planning, organization and monitoring process in the Health System; training and educational interventions on staff and exchange attitudes and behavior are necessary in order to offer a health care that is comprehensive, decent, human and with respect for the human rights.
Asunto(s)
Animales , Femenino , Humanos , Ratones , Antimetabolitos Antineoplásicos/farmacología , Ciclinas/metabolismo , Inhibidores Enzimáticos/metabolismo , Fenilacetatos/farmacología , Elementos sin Sentido (Genética) , Neoplasias de la Mama , Proteínas Quinasas Dependientes de Calcio-Calmodulina/metabolismo , División Celular/efectos de los fármacos , Quinasas Ciclina-Dependientes/genética , Quinasas Ciclina-Dependientes/metabolismo , Ciclinas/genética , Fibroblastos/citología , Fibroblastos/efectos de los fármacos , Fibroblastos/enzimología , Regulación Neoplásica de la Expresión Génica/fisiología , Ratones Noqueados , Fosforilación , Proteínas Serina-Treonina Quinasas/genética , Proteínas Serina-Treonina Quinasas/metabolismo , ARN Mensajero/metabolismo , Proteína de Retinoblastoma/metabolismo , Transducción de Señal/fisiología , Células Tumorales Cultivadas/citología , Células Tumorales Cultivadas/efectos de los fármacos , Células Tumorales Cultivadas/enzimología , Regulación hacia Arriba/efectos de los fármacosRESUMEN
The present study aimed to evaluate the influence of the following irrigating solutions on the microhardness of root canal dentin: 2% sodium hypochlorite (2NaOCl), 5% sodium hypochlorite (5NaOCl), super-oxidized water (400 ppm Sterilox - Sx) and 17% EDTA (E). Eighty roots from bovine incisors were randomly divided into 8 groups (n=10): 2NaOCl, 5NaOCl, Sx, and 2NaOCl + E, 5NaOCl + E, Sx + E (associated with E as final irrigant for 5 min), E solely and distilled water (dH2O) as the negative control. Root canal preparation was performed by hand instruments, using one of the irrigation protocols for 30 min. Then, 5 mm of the cervical root third were cut out from each sample and subjected to the Vickers microhardness test, at two points, one at approximately 500-1000 µm from the root canal lumen (distance 1), and the other at approximately 500-1000 µm from the external root surface (distance 2). Data were analyzed by Wilcoxon and Kruskal-Wallis tests at 5% significance level. Microhardness values at distance 1 were significantly lower than those at distance 2 for all groups, except 5NaOCl and 5NaOCl + E groups (p>0.05). EDTA showed the lowest microhardness values. However, no statistically significant difference was detected among groups at distance 1 and EDTA was significantly different only from Sx at distance 2. In conclusion, all tested solutions showed lower microhardness at the most superficial root canal dentin layer compared to the one found near the external root surface, except 5NaOCl and 5NaOCl + E; EDTA promoted lower microhardness values in comparison to Sterilox at this site.
O presente estudo teve como objetivo avaliar a influência das seguintes soluções irrigadoras na microdureza da dentina do canal radicular: hipoclorito de sódio a 2% (NaOCl2), hipoclorito de sódio a 5% (NaOCl5), água superoxidada (Sterilox(r) 400 ppm - Sx) e EDTA a 17% (E). Oitenta raízes de incisivos bovinos foram divididas aleatoriamente em 8 grupos (n=10): NaOCl2, NaOCl5, Sx e NaOCl2 + E, NaOCl5 + E, Sx + E (associados ao E como irrigante final por 5 min), E isolado e água destilada (H2Od), como controle negativo. O preparo dos canais radiculares foi realizado com instrumentos manuais, usando um dos protocolos de irrigação por 30 min. A seguir, 5 mm do terço cervical de cada amostra foram cortados perpendicularmente e submetidos ao teste de microdureza de Vickers, em dois pontos, um aproximadamente 500-1000 µm da luz do canal radicular (distância 1), e o outro aproximadamente 500-1000 µm da superfície externa da raiz (distância 2). Os dados foram analisados pelos testes de Wilcoxon e Kruskal-Wallis com um nível de significância de 5%. Os valores de microdureza na distância 1 foram significativamente menores do que na distância 2 para todos os grupos, exceto NaOCl5 e NaOCl5 +E (p>0,05). O EDTA mostrou os menores valores de microdureza. No entanto, não foi detectada diferença estatisticamente significativa entre os grupos na distância 1 e o EDTA foi significativamente diferente apenas do Sx na distância 2. Pode-se concluir que todas as soluções testadas mostraram menor microdureza na camada de dentina mais superficial do canal radicular em comparação aos valores encontrados próximo à superfície radicular externa, exceto NaOCl5 e NaOCl5 + E; o EDTA promoveu menor microdureza em comparação ao Sterilox(r) neste ponto.
Asunto(s)
Humanos , Antiinflamatorios no Esteroideos/farmacología , Antineoplásicos/farmacología , Carcinoma de Células Escamosas/tratamiento farmacológico , Neoplasias de la Boca/tratamiento farmacológico , Receptores Citoplasmáticos y Nucleares/metabolismo , Sulindac/análogos & derivados , Sulindac/farmacología , Factores de Transcripción/metabolismo , Apoptosis/efectos de los fármacos , Carcinoma de Células Escamosas/genética , Carcinoma de Células Escamosas/metabolismo , Ciclo Celular/efectos de los fármacos , División Celular/efectos de los fármacos , Inhibidores de la Ciclooxigenasa/farmacología , Cartilla de ADN/química , Citometría de Flujo , Técnicas para Inmunoenzimas , Isoenzimas/metabolismo , Proteínas de la Membrana , Neoplasias de la Boca/genética , Neoplasias de la Boca/metabolismo , Oligonucleótidos Antisentido/farmacología , Prostaglandina-Endoperóxido Sintasas/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , ARN Mensajero/metabolismo , Células Tumorales Cultivadas/efectos de los fármacos , Regulación hacia ArribaRESUMEN
This study compared the physicochemical properties and interfacial adaptation to canal walls of Endo-CPM-Sealer, Sealapex and Activ GP with the well-established AH Plus sealer. The following analyses were performed: radiopacity, pH variation and solubility using samples of each material and scanning electron microscopy of root-filled bovine incisors to evaluate the interfacial adaptation. Data were analyzed by the parametric and no-parametric tests (α=0.05). All materials were in accordance with the ANSI/ADA requirements for radiopacity. Endo-CPM-Sealer presented the lowest radiopacity values and AH Plus was the most radiopaque sealer (p=0.0001). Except for ActiV GP, which was acidic, all other sealers had basic chemical nature and released hydroxyl ions. Regarding solubility, all materials met the ANSI/ADA recommendations, with no statistically significant difference between the sealers (p=0.0834). AH Plus presented the best adaptation to canal walls in the middle (p=0.0023) and apical (p=0.0012) thirds, while the sealers Activ GP and Endo-CPM-Sealer had poor adaptation to the canal walls. All sealers, except for ActiV GP, were alkaline and all of them fulfilled the ANSI/ADA requirements for radiopacity and solubility. Regarding the interfacial adaptation, AH Plus was superior to the others considering the adaptation to the bovine root canal walls.
Este estudo comparou as propriedades físico-químicas e a adaptação interfacial às paredes do canal dos cimentos Endo-CPM-Sealer, Sealapex e Activ GP com o bem estabelecido cimento AH Plus. As seguintes análises foram realizadas: radiopacidade, variação de pH e de solubilidade utilizando amostras de cada material, e microscopia eletrônica de varredura utilizando incisivos bovinos obturados para avaliar a adaptação interfacial. Os dados foram analisados utilizando testes paramétricos e não-paramétricos (α=0,05). Todos os materiais estavam de acordo com os requerimentos da ANSI/ADA para radiopacidade, sendo que o Endo-CPM-Sealer apresentou os menores valores de radiopacidade e o AH Plus foi o cimento mais radiopaco (p=0,0001). Exceto o Activ GP, que foi ácido, todos os outros cimentos apresentaram natureza química básica e liberaram íons hidroxila. Com relação à solubilidade, todos os materiais estavam de acordo com as recomendações da ANSI /ADA, sem diferença significante entre os cimentos (p=0,0834). O AH Plus apresentou a melhor adaptação às paredes do canal nos terços médio (p=0,0023) e apical (p=0,0012), enquanto que os cimentos Activ GP e Endo-CPM-Sealer apresentaram uma pobre adaptação às paredes do canal. Em conclusão, todos os cimentos, exceto o Activ GP, foram alcalinos e todos preencheram os requerimentos da ANSI/ADA para radiopacidade e solubilidade. Com relação à adaptação interfacial, o AH Plus foi superior aos demais para adaptação às paredes do canal radicular de incisivos bovinos.
Asunto(s)
Animales , Femenino , Humanos , Ratones , Inhibidores de la Angiogénesis/farmacología , Antineoplásicos/farmacología , Neoplasias de la Mama/tratamiento farmacológico , Neoplasias de la Mama/patología , Dextranos/farmacología , Inhibidores de Crecimiento/farmacología , Células Tumorales Cultivadas/efectos de los fármacos , Células Tumorales Cultivadas/patología , Inhibidores de la Angiogénesis/uso terapéutico , Antineoplásicos/uso terapéutico , Medios de Cultivo Condicionados/farmacología , Relación Dosis-Respuesta a Droga , Dextranos/química , Dextranos/uso terapéutico , Endotelio Vascular/citología , Endotelio Vascular/efectos de los fármacos , Endotelio Vascular/fisiología , Inhibidores de Crecimiento/uso terapéutico , Ratones Endogámicos BALB C , Ratones Desnudos , Necrosis , Fenilacetatos/farmacología , Fenilacetatos/uso terapéutico , Ensayos Antitumor por Modelo de Xenoinjerto/estadística & datos numéricosRESUMEN
PURPOSE: Suppressor of cytokine signaling 7 (SOCS7) is a member of the SOCS family and is known to interact with phospholipase Cγ-1 (PLCγ-1), one of the insulin-like growth factor-I (IGF-I) receptor downstream molecules. In this study, we sought to observe the effect of knocking down SOCS7 gene on breast cancer cells in vitro growth and migration and to elucidate whether this involves IGF-I-PLCγ1 route using the PLCγ-1 blocker U73122. METHODS: Suitable breast cancer cells (MCF7 and MDA-MB-231) were transfected with anti-SOCS7 ribozymal transgene, to create sub-lines with SOCS7 knockdown verified by RT-PCR. The growth and migration of the cells were evaluated in the presence or absence of IGF-I and PLCγ-1 inhibitor using growth assay, scratch-wound and electrical cell impedance sensing (ECIS) migration assays. RESULTS: IGF-I treatment produced more pronounced influence on MCF7 growth and migration and on MDA-MB-231 migration when SOCS7 gene was knocked down in both lines (p < 0.05). The absence of IGF-I-induced growth response in MDA-MB-231 could be due to the intrinsic characteristics of these cells. PLCγ-1 pharmacological inhibition during their in vitro migration seemed to only occur when SOCS7 gene was knocked down. CONCLUSIONS: To the best of our knowledge, this is the first report of the SOCS7 regulatory role in IGF-I induced in vitro functions in ER-positive and ER-negative breast cancer cells. IGF-I treatment and SOCS7 loss have synergistically resulted in increased growth and migration of MCF7 and in increased migration of MDA-MB-231 cells. The migratory effects could be due to a precise anti-PLCγ-1 role.
Asunto(s)
Neoplasias de la Mama/genética , Técnicas de Silenciamiento del Gen , Factor I del Crecimiento Similar a la Insulina/farmacología , Proteínas Nucleares/genética , Proteínas Supresoras de la Señalización de Citocinas/genética , Humanos , Células Tumorales Cultivadas/efectos de los fármacosRESUMEN
OBJECTIVE: To determine whether curcumin reverses the multidrug resistance of human colon cancer cells in vitro and in vivo. METHODS: In a vincristine-resistant cell line of human colon cancer, the cell viability of curcumin-treated cells was determined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Rhodamine123 efflux was evaluated to detect P-glycoprotein transporter activity, and expression of the multidrug resistance protein 1 and survivin genes was analyzed by reverse transcription polymerase chain reaction and western blotting. In addition, xenograft mouse tumors were grown and treated with curcumin. The morphology of the xenografts was investigated by hematoxylin-eosin staining. The in vivo expression of the multidrug resistance gene and P-glycoprotein and survivin genes and proteins was observed using reverse transcription-polymerase chain reaction and western blotting, respectively. RESULTS: Curcumin was not obviously toxic to the vincristine-resistant human colon cancer cells at concentrations less than 25 µM, but the growth of cells was significantly inhibited. At concentrations greater than 25 µM, curcumin was toxic in a concentration-dependent manner. The sensitivity of cells to vincristine, cisplatin, fluorouracil, and hydroxycamptothecin was enhanced, intracellular Rhodamine123 accumulation was increased (p<0.05), and the expression of the multidrug resistance gene and P-glycoprotein were significantly suppressed (p<0.05). The combination of curcumin and vincristine significantly inhibited xenograft growth. The expression of the multidrug resistance protein 1 and survivin genes was significantly reduced in xenografts of curcumin-treated mice and mice treated with both curcumin and vincristine relative to control mice. CONCLUSION: Curcumin has strong reversal effects on the multidrug resistance of human colon carcinoma in vitro and in vivo.
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Antineoplásicos/farmacología , Neoplasias del Colon/tratamiento farmacológico , Curcumina/farmacología , Resistencia a Múltiples Medicamentos/efectos de los fármacos , Resistencia a Antineoplásicos/efectos de los fármacos , Animales , Supervivencia Celular/efectos de los fármacos , Neoplasias del Colon/patología , Femenino , Humanos , Ratones , Ratones Endogámicos BALB C , Células Tumorales Cultivadas/efectos de los fármacosRESUMEN
OBJECTIVE: To determine whether curcumin reverses the multidrug resistance of human colon cancer cells in vitro and in vivo. METHODS: In a vincristine-resistant cell line of human colon cancer, the cell viability of curcumin-treated cells was determined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Rhodamine123 efflux was evaluated to detect P-glycoprotein transporter activity, and expression of the multidrug resistance protein 1 and survivin genes was analyzed by reverse transcription polymerase chain reaction and western blotting. In addition, xenograft mouse tumors were grown and treated with curcumin. The morphology of the xenografts was investigated by hematoxylin-eosin staining. The in vivo expression of the multidrug resistance gene and P-glycoprotein and survivin genes and proteins was observed using reverse transcription-polymerase chain reaction and western blotting, respectively. RESULTS: Curcumin was not obviously toxic to the vincristine-resistant human colon cancer cells at concentrations less than 25 μM, but the growth of cells was significantly inhibited. At concentrations greater than 25 μM, curcumin was toxic in a concentration-dependent manner. The sensitivity of cells to vincristine, cisplatin, fluorouracil, and hydroxycamptothecin was enhanced, intracellular Rhodamine123 accumulation was increased (p<0.05), and the expression of the multidrug resistance gene and P-glycoprotein were significantly suppressed (p<0.05). The combination of curcumin and vincristine significantly inhibited xenograft growth. The expression of the multidrug resistance protein 1 and survivin genes was significantly reduced in xenografts of curcumin-treated mice and mice treated with both curcumin and vincristine relative to control mice. CONCLUSION: Curcumin has strong reversal effects on the multidrug resistance of human colon carcinoma in vitro and in vivo. .
Asunto(s)
Animales , Femenino , Humanos , Ratones , Antineoplásicos/farmacología , Neoplasias del Colon/tratamiento farmacológico , Curcumina/farmacología , Resistencia a Múltiples Medicamentos/efectos de los fármacos , Resistencia a Antineoplásicos/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Neoplasias del Colon/patología , Ratones Endogámicos BALB C , Células Tumorales Cultivadas/efectos de los fármacosRESUMEN
INTRODUCTION: The Egr-1 protein is a transcriptional factor responsive to early growth. Transcriptional regulation of the promoter has been described like responsive to physical stress, osmotic changes, and cellular growth marker. However, there is no report about the pharmacological effect on the transcriptional regulation in gliomas. Hereby we report the modulation of the Egr-1 promoter transcriptional activity induced by the Granulocytes Macrophages Colony Stimulating Factor (GM-CSF) and steroid drugs in human glioma cells (CH235-GM Grade II, U373-GM Grade III, D54-GM Grade IV) using a reporter system transduced by a recombinant adenoviral vector AdEgr-1/luc7. METHODS: Human glioma cells shows with different malignity grade (CH235-GM Grado II; U373-GM Grado III; D54-GM Grado IV) were transduced with no replicative adenoviral vector AdEgr-1/Luc7 and exposed to drugs as progesterone, ß-estradiol and betametasone, and GM-CSF. Transcriptional activity of the egr-1 promoter was quantified by Luciferase reporter gene, cloned downstream to the tata box. Luciferase activity was quantified from whole cell proteins using luminometry assays. RESULTS: U373-GM cell line with GM-CSF, shows an increment on transcriptional activity of Egr-1 promoter, also in endogen way. U373-GM showed a positive regulation of Egr-1, with steroid drugs on the times analyzed. Steroid drugs as progesterone, ß-estradiol and betametasone, shows a pleiotropic behavior on CH235-GM and D54-GM, glioma cell lines. CONCLUSIONS: Inhibition or activation response of Egr-1 promoter shows new framework to explore a mechanism of action of steroid drugs on genetic and epigenetic regulation on tumoral process.
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Betametasona/farmacología , Proteína 1 de la Respuesta de Crecimiento Precoz/farmacología , Glioma/patología , Glucocorticoides/farmacología , Factor Estimulante de Colonias de Granulocitos y Macrófagos/farmacología , Humanos , Células Tumorales Cultivadas/efectos de los fármacosRESUMEN
Two decades ago, it was hypothesized that antidepressants could alter the course of neoplastic diseases. However, contradictory findings indicated that antidepressants could either have carcinogenic properties or improve the disease outcome. Intriguingly, controversial results were reported on the action of antidepressant drugs on immune function. Further hypotheses proposed that antidepressants could indirectly affect the cancer prognosis through the modulation of antitumor activity. Here we review the literature in order to elucidate the influence of antidepressants on cancer and immunity.
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Antidepresivos/efectos adversos , Antidepresivos/inmunología , Antidepresivos/metabolismo , Inmunidad Celular/efectos de los fármacos , Metástasis de la Neoplasia , Neoplasias/inducido químicamente , Neoplasias/epidemiología , Transducción de Señal/efectos de los fármacos , Animales , Apoptosis/efectos de los fármacos , Ciclo Celular/efectos de los fármacos , Humanos , Modelos Biológicos , Nitrosación/efectos de los fármacos , Oxidación-Reducción/efectos de los fármacos , Factores de Riesgo , Transducción de Señal/inmunología , Células Tumorales Cultivadas/efectos de los fármacosRESUMEN
Apatone™, a combination of menadione (2-methyl-1,4-naphthoquinone, VK3) and ascorbic acid (vitamin C, VC) is a new strategy for cancer treatment. Part of its effect on tumor cells is related to the cellular pro-oxidative imbalance provoked by the generation of hydrogen peroxide (H2O2) through naphthoquinone redox cycling. In this study, we attempted to find new naphthoquinone derivatives that would increase the efficiency of H2O2 production, thereby potentially increasing its efficacy for cancer treatment. The presence of an electron-withdrawing group in the naphthoquinone moiety had a direct effect on the efficiency of H2O2 production. The compound 2-bromo-1,4-naphthoquinone (BrQ), in which the bromine atom substituted the methyl group in VK3, was approximately 10- and 19-fold more efficient than VK3 in terms of oxygen consumption and H2O2 production, respectively. The ratio [H2O2]produced / [naphthoquinone]consumed was 68 ± 11 and 5.8 ± 0.2 (µM/µM) for BrQ and VK3, respectively, indicating a higher efficacy of BrQ as a catalyst for the autoxidation of ascorbic acid. Both VK3 and BrQ reacted with glutathione (GSH), but BrQ was the more effective substrate. Part of GSH was incorporated into the naphthoquinone, producing a nucleophilic substitution product (Q-SG). The depletion of BrQ by GSH did not prevent its redox capacity since Q-SG was also able to catalyze the production of reactive oxygen species. VK3/VC has already been submitted to clinical trials for the treatment of prostate cancer and has demonstrated promising results. However, replacement of VK3 with BrQ will open new lines of investigation regarding this approach to cancer treatment.
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Humanos , Antineoplásicos/farmacología , Ácido Ascórbico/farmacología , Peróxido de Hidrógeno/metabolismo , Naftoquinonas/farmacología , Especies Reactivas de Oxígeno , Antineoplásicos/química , Ácido Ascórbico/química , Combinación de Medicamentos , Sustitución de Medicamentos , Naftoquinonas/química , Oxidación-Reducción/efectos de los fármacos , Consumo de Oxígeno/efectos de los fármacos , Relación Estructura-Actividad , Células Tumorales Cultivadas/efectos de los fármacos , /química , /farmacologíaRESUMEN
Apatone™, a combination of menadione (2-methyl-1,4-naphthoquinone, VK3) and ascorbic acid (vitamin C, VC) is a new strategy for cancer treatment. Part of its effect on tumor cells is related to the cellular pro-oxidative imbalance provoked by the generation of hydrogen peroxide (H2O2) through naphthoquinone redox cycling. In this study, we attempted to find new naphthoquinone derivatives that would increase the efficiency of H2O2 production, thereby potentially increasing its efficacy for cancer treatment. The presence of an electron-withdrawing group in the naphthoquinone moiety had a direct effect on the efficiency of H2O2 production. The compound 2-bromo-1,4-naphthoquinone (BrQ), in which the bromine atom substituted the methyl group in VK3, was approximately 10- and 19-fold more efficient than VK3 in terms of oxygen consumption and H2O2 production, respectively. The ratio [H2O2]produced / [naphthoquinone]consumed was 68 ± 11 and 5.8 ± 0.2 (µM/µM) for BrQ and VK3, respectively, indicating a higher efficacy of BrQ as a catalyst for the autoxidation of ascorbic acid. Both VK3 and BrQ reacted with glutathione (GSH), but BrQ was the more effective substrate. Part of GSH was incorporated into the naphthoquinone, producing a nucleophilic substitution product (Q-SG). The depletion of BrQ by GSH did not prevent its redox capacity since Q-SG was also able to catalyze the production of reactive oxygen species. VK3/VC has already been submitted to clinical trials for the treatment of prostate cancer and has demonstrated promising results. However, replacement of VK3 with BrQ will open new lines of investigation regarding this approach to cancer treatment.
Asunto(s)
Antineoplásicos/farmacología , Ácido Ascórbico/farmacología , Peróxido de Hidrógeno/metabolismo , Naftoquinonas/farmacología , Especies Reactivas de Oxígeno , Antineoplásicos/química , Ácido Ascórbico/química , Combinación de Medicamentos , Sustitución de Medicamentos , Humanos , Naftoquinonas/química , Oxidación-Reducción/efectos de los fármacos , Consumo de Oxígeno/efectos de los fármacos , Relación Estructura-Actividad , Células Tumorales Cultivadas/efectos de los fármacos , Vitamina K 3/química , Vitamina K 3/farmacologíaRESUMEN
OBJECTIVE: To evaluate the cytotoxic effect of a hexane extract of Cassia alata leaves in A549 lung cancer cells. METHOD: Parental A549 lung cancer cells were exposed to various concentrations (100"180 µg/ml) of Cassia alata leaf extract for 24 hours. Following treatment, the cells were evaluated using the 3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay to determine the cytotoxic effect of the extract. Caspase 8, 3 and 9 negative A549 cells were also prepared using lentiviral based shRNA knockdown of the caspase 8, 3 and 9 genes, respectively. The cytotoxic effect of Cassia alata leaf extract was then evaluated in these knockdown cells using the MTT assay. Chemical analysis was performed on the extract using high performance liquid chromatography (HPLC). RESULTS: Cassia alata extract was cytotoxic in parental and caspase-9 negative, but not caspase 3 and 8 negative A549 cells. The IC50 values were 143 µg/ml and 145 µg/ml in parental and caspase 9 negative A549 cells respectively. The flavanoid kaempferol was identified as a constituent of Cassia alata leaf extract. CONCLUSIONS: Cassia alata produces cytotoxicity in A549 cancer cells that is mediated by caspase 8 activation. This effect may be attributable to kaempferol.
OBJETIVO: Evaluar el efecto citotóxico de un extracto de hexano de hojas de Cassia alata en las células A549 del cáncer pulmonar. MÉTODO: Células A549 parentales del cáncer pulmonar fueron expuestas a varias concentraciones (100-180 µg/ml) de un extracto de la hoja de Cassia alatadurante 24 horas. Tras el tratamiento, las células fueron evaluadas usando el ensayo de bromuro de 3-(4,5-dimetiltiazol-2-il)-2,5-difeniltetrazolio (MTT) a fin de determinar el efecto citotóxico del extracto. También se prepararon células A549 negativas caspasa 8, 3 y 9 mediante silenciamiento génico vía ARN (shRNA knockdown) de los genes de las caspasas 8, 3 y 9 respectivamente, sobre la base de la inserción de vectores lentivirales. Entonces, usando un ensayo MTT se procedió a evaluar el efecto citotóxico del extracto de hojas de Cassia alataen éstas células genéticamente modificadas. Se realizó un análisis químico del extracto utilizando cromatografía líquida de alta eficacia. (HPLC). RESULTADOS: El extracto de Cassia alata resultó ser citotóxico en las células A549 negativas parentales y caspasa 9, pero no en las negativas caspasa 3 y 8. Los valores de IC50 fueron 143 µg/ml y 145 µg/ml en las células A549 negativas parentales y caspasa 9 respectivamente. El flavonol kaempferol fue identificado como un constituyente del extracto de las hojas de Cassia alata. CONCLUSIONES: La Cassia alata produce citotoxicidad en las células cancerosas A549, mediada por la activación de la caspasa 8. Este efecto puede ser atribuido al kaempferol.