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Article in English | WPRIM | ID: wpr-887724


Objective@#Although benzene is a confirmed environmental carcinogen, the mechanism of its carcinogenicity remains largely unclear. The suggested oncogene, miR-221, is elevated and plays important roles in various tumors, but its role in benzene-induced carcinogenesis remains unknown.@*Methods@#In the present study, we constructed hydroquinone (HQ, a representative metabolite of benzene with biological activity)-transformed malignant cell line (16HBE-t) and analyzed the level of miR-221 in it with qRT-PCR. Exosomes from 16HBE-t cells incubated with or without an miR-221 inhibitor were isolated by ultracentrifugation, characterized by transmission electron microscopy and laser scanning confocal microscope, and then transfected into 16HBE cells. The effects of exosomal miR-221 on apoptosis induced by HQ in recipient cells were determined using flow cytometry.@*Results@#The amount of miR-221 in 16HBE-t was significantly increased compared with controls. When recipient cells ingested exosomes derived from 16HBE-t, miR-221 was increased, and apoptosis induced by HQ was inhibited. Blocking miR-221 in 16HBE-t using an inhibitor did not significantly alter miR-221 or apoptosis in recipient cells.@*Conclusion@#Exosomal miR-221 secreted by 16HBE-t inhibits apoptosis induced by HQ in normal recipient cells.

Apoptosis , Bronchi/cytology , Cell Line, Transformed , Epithelial Cells , Exosomes , Humans , Hydroquinones , MicroRNAs
Braz. dent. j ; 29(4): 359-367, July-Aug. 2018. tab, graf
Article in English | LILACS | ID: biblio-974167


Abstract The aim of this study was to evaluate the antifungal activity of Terpinen-4-ol associated with nystatin, on single and mixed species biofilms formed by Candida albicans and Candida tropicalis, as well as the effect of terpinen-4-ol on adhesion in oral cells and the enzymatic activity. The minimum inhibitory concentrations and minimum fungicide concentrations of terpinen-4-ol and nystatin on Candida albicans and Candida tropicalis were determined using the microdilution broth method, along with their synergistic activity ("checkerboard" method). Single and mixed species biofilms were prepared using the static microtiter plate model and quantified by colony forming units (CFU/mL). The effect of Terpinen-4-ol in adhesion of Candida albicans and Candida tropicalis in coculture with oral keratinocytes (NOK Si) was evaluated, as well as the enzymatic activity by measuring the size of the precipitation zone, after the growth agar to phospholipase, protease and hemolysin. Terpinen-4-ol (4.53 mg mL-1) and nystatin (0.008 mg mL-1) were able to inhibit biofilms growth, and a synergistic antifungal effect was showed with the drug association, reducing the inhibitory concentration of nystatin up to 8 times in single biofilm of Candida albicans, and 2 times in mixed species biofilm. A small decrease in the adhesion of Candida tropicalis in NOK Si cells was showed after treatment with terpinen-4-ol, and nystatin had a greater effect for both species. For enzymatic activity, the drugs showed no action. The effect potentiated by the combination of terpinen-4-ol and nystatin and the reduction of adhesion provide evidence of its potential as an anti-fungal agent.

Resumo O objetivo desse estudo foi avaliar a atividade antifúngica do Terpinen4-ol associado à nistatina em biofilmes simples e misto, formados por Candida albicans e Candida tropicalis, bem como o efeito do terpinen-4-ol na adesão em células orais e atividade enzimática. As concentrações inibitórias mínimas e as concentrações fungicidas mínimas do terpinen-4-ol e da nistatina em Candida albicans e Candida tropicalis foram determinadas pelo método de microdiluição em caldo, juntamente com a atividade sinérgica (método do tabuleiro de "xadrez"). Biofilmes simples e misto foram preparados usando o modelo de placa de microtitulação estática e quantificados por unidades formadoras de colônias (CFU/mL). O efeito do Terpinen-4-ol na adesão de Candida albicans e Candida tropicalis em co-cultura com queratinócitos orais (NOK Si) foi avaliado, bem como a atividade enzimática, medindo o tamanho da zona de precipitação, após o crescimento em ágar fosfolipase, protease e hemolisina. O terpinen-4-ol (4.53 mg mL-1) e a nistatina (0,008 mg mL-1) conseguiram inibir o crescimento de biofilmes e um efeito antifúngico sinérgico foi demonstrado com a associação de fármaco, reduzindo a concentração inibidora de nistatina até 8 vezes em biofilme simpes de Candida albicans e 2 vezes em biofilme misto. Uma pequena diminuição na adesão de Candida tropicalis em células NOK Si foi mostrada após o tratamento com terpinen-4-ol e a nistatina teve um efeito maior para ambas as espécies. Para a atividade enzimática, as drogas não apresentaram ação. O efeito potencializado pela combinação de terpinen-4-ol e nistatina e a redução de adesão evidenciam seu potencial como agente anti-fúngico.

Terpenes/pharmacology , Candida albicans/drug effects , Nystatin/pharmacology , Biofilms/drug effects , Candida tropicalis/drug effects , Antifungal Agents/pharmacology , Cell Line, Transformed , Microbial Sensitivity Tests , Drug Synergism
Braz. j. med. biol. res ; 51(12): e7862, 2018. graf
Article in English | LILACS | ID: biblio-974259


Although the effects of low-intensity pulsed ultrasound (LIPUS) on diverse cell types have been fully studied, the functional role of LIPUS in keratinocytes remains poorly understood. This study aimed to investigate the effects of LIPUS on proliferation and migration of HaCaT cells as well as the regulatory mechanisms associated with signaling pathways. Human HaCaT cells were exposed or not to LIPUS, and cell proliferation and migration were measured by BrdU incorporation assay and Transwell assay, respectively. Expression of proteins associated with proliferation and migration was evaluated by western blot analysis. Expression of key kinases in the PI3K/AKT and JNK pathways was also evaluated by western blot analysis. Effects of LIPUS on the PI3K/AKT and JNK pathways, and whether LIPUS affected HaCaT cells via these two pathways were finally explored. When the parameter of LIPUS (number of cycles) was set at 300, cell viability was the highest after LIPUS stimulation. We then found that the percentage of BrdU positive cells was enhanced by LIPUS, along with up-regulation of cyclinD1, CDK6, CDK4, and VEGF. LIPUS promoted migration, as well as up-regulation of MMP-2 and MMP-9. Phosphorylation levels of key kinases in the PI3K/AKT and JNK pathways were increased by LIPUS. Inhibition of either PI3K/AKT pathway or JNK pathway attenuated effects of LIPUS on HaCaT cells, and co-inhibition of these two pathways showed augmented effects. LIPUS promoted proliferation and migration of HaCaT cells through activating the PI3K/AKT and JNK pathways.

Keratinocytes/radiation effects , Cell Movement/radiation effects , Phosphatidylinositol 3-Kinases/radiation effects , MAP Kinase Signaling System/radiation effects , Cell Proliferation/radiation effects , Ultrasonic Waves , Bromodeoxyuridine , Cell Line, Transformed , Signal Transduction/radiation effects , Keratinocytes/metabolism , Up-Regulation , Cell Survival/radiation effects , Blotting, Western , Reproducibility of Results , Analysis of Variance , Phosphatidylinositol 3-Kinases/metabolism , Matrix Metalloproteinase 2/metabolism , Matrix Metalloproteinase 9/metabolism
Clinics ; 73(supl.1): e539s, 2018. graf
Article in English | LILACS | ID: biblio-952838


Infection with high oncogenic risk human papillomavirus types is the etiological factor of cervical cancer and a major cause of other epithelial malignancies, including vulvar, vaginal, anal, penile and head and neck carcinomas. These agents affect epithelial homeostasis through the expression of specific proteins that deregulate important cellular signaling pathways to achieve efficient viral replication. Among the major targets of viral proteins are components of the DNA damage detection and repair machinery. The activation of many of these cellular factors is critical to process viral genome replication intermediates and, consequently, to sustain faithful viral progeny production. In addition to the important role of cellular DNA repair machinery in the infective human papillomavirus cycle, alterations in the expression and activity of many of its components are observed in human papillomavirus-related tumors. Several studies from different laboratories have reported the impact of the expression of human papillomavirus oncogenes, mainly E6 and E7, on proteins in almost all the main cellular DNA repair mechanisms. This has direct consequences on cellular transformation since it causes the accumulation of point mutations, insertions and deletions of short nucleotide stretches, as well as numerical and structural chromosomal alterations characteristic of tumor cells. On the other hand, it is clear that human papillomavirus-transformed cells depend on the preservation of a basal cellular DNA repair activity level to maintain tumor cell viability. In this review, we summarize the data concerning the effect of human papillomavirus infection on DNA repair mechanisms. In addition, we discuss the potential of exploiting human papillomavirus-transformed cell dependency on DNA repair pathways as effective antitumoral therapies.

Humans , Papillomaviridae/genetics , Papillomavirus Infections/virology , DNA Repair , Neoplasms/virology , Papillomaviridae/physiology , Virus Replication , Cell Line, Transformed/virology , Cell Survival/genetics , Genomic Instability/genetics , Neoplasms/therapy
Neuroscience Bulletin ; (6): 1037-1046, 2018.
Article in English | WPRIM | ID: wpr-775486


Autophagy is an evolutionarily-conserved self-degradative process that maintains cellular homeostasis by eliminating protein aggregates and damaged organelles. Recently, vesicle-associated membrane protein-associated protein B (VAPB), which is associated with the familial form of amyotrophic lateral sclerosis, has been shown to regulate autophagy. In the present study, we demonstrated that knockdown of VAPB induced the up-regulation of beclin 1 expression, which promoted LC3 (microtubule-associated protein light chain 3) conversion and the formation of LC3 puncta, whereas overexpression of VAPB inhibited these processes. The regulation of beclin 1 by VAPB was at the transcriptional level. Moreover, knockdown of VAPB increased autophagic flux, which promoted the degradation of the autophagy substrate p62 and neurodegenerative disease proteins. Our study provides evidence that the regulation of autophagy by VAPB is associated with the autophagy-initiating factor beclin 1.

Autophagy , Physiology , Beclin-1 , Genetics , Metabolism , Cell Line, Transformed , Gene Expression Regulation , Genetics , Green Fluorescent Proteins , Genetics , Metabolism , Humans , Microtubule-Associated Proteins , Genetics , Metabolism , R-SNARE Proteins , Genetics , Metabolism , RNA, Messenger , Metabolism , RNA, Small Interfering , Genetics , Metabolism , RNA-Binding Proteins , Genetics , Metabolism , Transfection
Braz. dent. j ; 28(1): 57-64, Jan.-Feb. 2017. tab, graf
Article in English | LILACS | ID: biblio-839113


Abstract This study aimed to evaluate the cytotoxicity of a calcium aluminate cement (EndoBinder) containing different radiopacifiers, Bi2O3, ZnO or ZrO2, compared with Mineral Trioxide Aggregate (MTA). According to ISO 10993-12:2012 (E) recommendations, 0.2 g of each cement were applied in transwell inserts and placed in 24-well culture plates containing 1 mL of culture medium (DMEM). After 24 h of incubation, the extracts (DMEM containing components released from the cements) were applied to immortalized odontoblast-like MDPC-23 cells. Cell viability (MTT test), alkaline phosphatase activity (ALP), total protein production and cell morphology (Scanning Electron Microscopy - SEM) were evaluated. The volume of 50 µL of extract was used to determine the chemical elements released by the cements using Energy Dispersive Spectroscopy (EDS). The following groups were established (n=6): NC - negative control (without treatment); EB - EndoBinder without radiopacifier; EBBO - EndoBinder+Bi2O3; EBZnO - EndoBinder+ZnO; EBZrO - EndoBinder+ZrO2 and WMTA - White MTA. Data were subjected to statistical analysis (Kruskal-Wallis test, level of significance=5%). Cells exposed to the different versions of EndoBinder presented small reduction in viability, total protein production and ALP activity, with values similar to the NC and WMTA groups (p>0.05). Different elements (C, O, Na, Al, P, Si, Cl, Bi, K) released by the cements were detected in the extracts. However, the cells had no significant changes in their morphology. EndoBinder and MTA did not affect negatively the metabolism of the odontoblastic-like cells, showing it to be cytocompatible, irrespective of the used radiopacifier.

Resumo Este estudo avaliou a citotoxicidade de um cimento de aluminato de cálcio (EndoBinder) contendo diferentes radiopacificadores, Bi2O3, ZnO ou ZrO2, comparativamente ao trióxido mineral agregado (MTA). Seguindo a norma ISO 10993-12:2012 (E), 0,2 g de cada cimento foi aplicada em insertos transwell, que foram colocados em placas de cultura de 24 wells contendo 1 mL de meio de cultura (DMEM). Após 24 h de incubação, os extratos (DMEM contendo componentes liberados dos cimentos) foram aplicados sobre células pulpares imortalizadas MDPC-23. Viabilidade celular (teste de MTT), atividade da fosfatase alcalina (ALP), produção de proteína total e a morfologia das células (Microscópio Eletrônico de Varredura - MEV) foram avaliadas. Um volume de 50 µL do extrato foi utilizado para determinar, através de Espectroscopia de Energia Dispersiva (EDS), os elementos químicos liberados pelos cimentos. Os seguintes grupos foram estabelecidos (n=6): NC - controle negativo (sem tratamento); EB - EndoBinder sem radiopacificador; EBBO - EndoBinder+Bi2O3; EBZnO - EndoBinder+ZnO; EBZrO - EndoBinder+ZrO2 e WMTA - MTA branco. Os dados foram submetidos à análise estatística (teste de Kruskal-Wallis, nível de significância=5%). Células expostas às diferentes versões de EndoBinder apresentaram pequena redução na viabilidade, produção de proteína total e atividade da ALP, com valores semelhantes aos grupos NC e WMTA (p>0,05). Diversos elementos (C, O, Na, Al, P, Si, Cl, Bi, K) liberados pelos cimentos foram detectados nos extratos. Entretanto, as células não apresentaram alterações significativas em sua morfologia. EndoBinder e MTA, não afetaram negativamente o metabolismo das células odontoblastóides, mostrando-se citocompatíveis, independente do radiopacificador utilizado.

Animals , Calcium Compounds/toxicity , Aluminum Compounds/toxicity , Dental Cements/toxicity , Oxides/toxicity , Spectrometry, X-Ray Emission , Cell Line, Transformed , Microscopy, Electron, Scanning , Cell Survival/drug effects , Silicates/toxicity , Drug Combinations
Article in Chinese | WPRIM | ID: wpr-300822


To discover the nephroprotective substances from Huangkui capsule.The components of Huangkui capsule were isolated by preparative liquid chromatography, and the active components were screened by LC/MS and identified. The adriamycine-injured HK-2 cells were treated with various active components with different concentrations, and the malonaldehyde (MDA) content, adenosine triphosphate (ATP) level and mitochondrial oxygen consumption rate were measured to verify the protective activity of the compounds.Four active components in Huangkui capsule were identified to exert nephroprotective effects. Fifteen flavanoids from these four components were tentatively identified by LC/MS, and hyperin, myricetin, quercetin, rutin and isoquercetin were confirmed. Hyperin, myricetin quercetin and rutin showed dose-dependent protective effects on injured HK-2 cells. Espacially, hyperin significantly reduced MDA content, quercetin and rutin significantly increased ATP level, and myricetin significantly increased mitochondrial oxygen consumption rate.Hyperin, myricetin, querctein and rutin might be the potential nephroprotective compounds in Huangkui capsule, their effects may be related to the inhibition of lipid peroxidation and the alleviation of mitochondrial damage.

Abelmoschus , Chemistry , Adenosine Triphosphate , Metabolism , Cell Line, Transformed , Chromatography, Liquid , Doxorubicin , Drugs, Chinese Herbal , Epithelial Cells , Flavonoids , Pharmacology , Kidney Diseases , Drug Therapy , Kidney Tubules, Proximal , Lipid Peroxidation , Malondialdehyde , Metabolism , Mass Spectrometry , Mitochondria , Oxygen Consumption , Protective Agents , Chemistry , Pharmacology , Quercetin , Pharmacology , Rutin , Pharmacology
Braz. dent. j ; 27(2): 160-168, Mar.-Apr. 2016. tab, graf
Article in English | LILACS | ID: lil-778327


Abstract The aim of this study was to evaluated the root surfaces modifications resulted by application of different chemicals agents, and their influence on the fibrin network and fibroblasts attachment. From 96 anterior mandibular human extracted incisor teeth, 192 dentin blocks of buccal and lingual surface were obtained and randomly divided into 6 groups: Cont- control group, which received no treatment; Root surface scaling and root planing (Srp); Citric acid-Srp; EDTA-Srp; Tetracycline capsule-Srp; Tetracycline gel-Srp. After dentin treatments the specimens were analyzed as follows: 1) demineralization level and residues of the product by scanning electron microscopy (SEM); 2) adhesion of blood components after 20 min of surface treatment by SEM; 3) fibroblast attachment after 24 h by SEM; 4) cell metabolism after 24 h by MTT assay. Data were analyzed using Fisher Exact, One-way ANOVA test followed by Dunn's test, Tukey test and Dunnett test (α=0.05). Citric acid, EDTA and Tetracycline gel resulted in adequate demineralization with no completely smear layer and smear plug removal on root dentin surface. Tetracycline capsule produced great tetracycline residues with several demineralization areas. Tetracycline gel and EDTA groups presented more fibroblast fixation than other experimental groups. The highest mean blood clot adhesion score was observed in roots treated with tetracycline gel. EDTA and Tetracycline gel surface treatment removed the smear layer over dentin surface and promoted adhesion of fibrin network and fibroblast cells attachment.

Resumo O objetivo deste estudo foi avaliar modificações nas superfícies radiculares sofridas pela aplicação de diferentes agentes químicos, e sua influência sobre a rede de fibrina e adesão de fibroblastos. A partir de 96 incisivos inferiores humanos, 192 blocos de dentina das superfícies vestibular e lingual foram obtidos e divididos aleatoriamente em 6 grupos: Cont-grupo controle, não recebeu nenhum tratamento; Raspagem e alisamento radicular (RAR); Ácido cítrico-SRP; EDTA-SRP; Tetraciclina em cápsula-SRP; Tetraciclina gel-SRP. Após o tratamento da dentina as espécimes foram analisadas: 1, nível de desmineralização e resíduos do produto por microscopia eletrônica de varredura (MEV); 2, adesão dos componentes sanguineos após 20 min na tratamento de superfície por SEM; 3, adesão de fibroblastos após 24h por SEM; 4, o metabolismo celular após 24 h por ensaio MTT. Os dados foram analisados por Fisher Exact, teste one-way ANOVA, seguido pelo teste de Dunn, teste de Tukey e teste de Dunnett (α = 0,05). O ácido cítrico, EDTA e gel tetraciclina resultaram na adequada desmineralização sem remoção completa de camada de smear layer e smear plug sobre a superfície da dentina radicular. Cápsula de tetraciclina produziu grandes resíduos de tetraciclina com várias áreas de desmineralização. Os grupos Gel de tetraciclina e EDTA apresentaram maior adesão de fibroblastos do que os demais grupos experimentais. O maior score de adesão de coágulo sanguineo foi observado nas superfícies tratadas com gel de tetraciclina. EDTA e Gel de tetraciclina removeram a camada de smear layer sobre a superfície da dentina e promoveu adesão da rede de fibrina e de fibroblastos.

Humans , Periodontium , Tooth Root/drug effects , Cell Line, Transformed , In Vitro Techniques , Microscopy, Electron, Scanning
Article in Chinese | WPRIM | ID: wpr-239572


To observe the effect of uniform and shift rotation culture on the formation and activity of the alginate-chitosan (AC) microencapsulated HepLL immortalized human hepatocytes and HepG2 cells aggregates.AC microcapsulated HepG2 and HepLL cells were randomly divided into two groups. Each group was divided into 3 subgroups according to uniform and shift rotation culture.The size and number of aggregates were observed and measured under laser confocal microscopy and inverted microscope dynamically. The amount of albumin synthesis was detected by ELISA, the clearance of ammonia was detected by colorimetry, and diazepam conversion function was detected by high performance liquid chromatography (HPLC).On day 6, 8, 10, 12, 14 and 16, the number and size of the aggregates, albumin synthesis, diazepam clearance and ammonium clearance increased significantly in shift rotation culture group than in uniform group (all<0.01). The albumin synthesis, diazepam clearance, and ammonium clearance in the microencapsulated HepLL groups were significantly higher than those of HepG2 cells at any time (all<0.01).Shift rotation culture can significantly promote the formation and increase the activity of AC microencapsulated HepLL and HepG2 aggregates, and HepLL cells may be more suitable for bioartificial liver than HepG2.

Albumins , Metabolism , Alginates , Ammonia , Metabolism , Animals , Cell Aggregation , Physiology , Cell Culture Techniques , Methods , Cell Line, Transformed , Physiology , Chitosan , Diazepam , Metabolism , Glucuronic Acid , Hep G2 Cells , Cell Biology , Physiology , Hepatocytes , Cell Biology , Physiology , Hexuronic Acids , Humans , Liver, Artificial , Rotation
Arch. argent. pediatr ; 113(5): 411-418, oct. 2015. graf, tab
Article in Spanish | LILACS | ID: lil-757062


Introducción. Existen evidencias de la asociación de determinantes sociales con la salud infantil. Objetivo. Identificar características sociodemográficas asociadas a desigualdades en la salud infantil y evaluar el efecto acumulado sobre la salud de factores de riesgo basados en estas características. Población y métodos. Evaluamos niños de 4-13 años, de Bariloche, entre junio de 2008 y mayo de 2009. Características sociodemográficas consideradas: nivel socioeconómico, educación materna, embarazo adolescente, cobertura médica, inseguridad y hábitos familiares. Valoramos la percepción parental de la salud física y socioemocional, el estado nutricional y la salud bucal en relación con dichas características y con la acumulación de factores de riesgo. Utilizamos encuesta, antropometría y examen bucal. Resultados. Participaron 180 escolares. El nivel educativo materno se asoció con la salud física, socioemocional y bucal del niño. El porcentaje de niños con piezas faltantes o caries fue 77% entre aquellos cuyas madres, como máximo, habían completado el primario, comparado con 13% entre aquellos cuyas madres habían completado estudios terciarios/universitarios. La posibilidad de percepción de salud física y socioemocional no óptima aumentó con cada factor de riesgo 1,8 y 1,4 veces, respectivamente, y la posibilidad de caries o piezas faltantes se duplicó con cada factor de riesgo adicional. El 27,3% de los escolares presentó sobrepeso y el 8,7%, obesidad, y no se encontró asociación con características sociodemográficas. Conclusiones. El bajo nivel socioeconómico familiar y educativo materno se asoció con una mayor prevalencia de resultados de salud desfavorables. Múltiples factores de riesgo tienen un efecto acumulado sobre la percepción parental de la salud física y socioemocional y la salud bucal.

Introduction. There is evidence of an association between social determinants and child health. Objective. To identify sociodemographic characteristics related to child health inequalities and to analize the cumulative effect on health of risk factors based on these characteristics. Population and Methods. We evaluated 4-13 year-old children in Bariloche between June 2008 and May 2009. The following sociodemographic characteristics were taken into account: socioeconomic level, maternal education, adolescent pregnancy, medical coverage, unsafeness, and family habits. We assessed parental perception of physical, and social and emotional health, nutritional status and oral health in relation to these characteristics and the accumulation of risk factors. We used survey, anthropometry and oral examination. Results. One hundred and eighty students participated. The level of maternal education was associated with the child's physical, social and emotional, and oral health. The percentage of children with missing teeth or cavities reached 77% among those whose mothers had, at most, completed primary school, compared to 13% among those whose mothers had completed tertiary school or university. The possibility of perceiving a non-optimal physical, and social and emotional health increased 1.8 and 1.4 times with each risk factor, respectively, and the possibility of having missing teeth or cavities was twice as much with each additional risk factor. Overweight and obesity was observed in 27.3% and 8.7% of students, respectively, and no relationship was found with sociodemographic characteristics. Conclusions. A low family socioeconomic level and a low maternal education level were associated with a higher prevalence of unfavorable health outcomes. Multiple risk factors have an cumulative effect on parental perception of physical, social and emotional, and oral health.

Humans , Cell Transformation, Viral/genetics , Gene Expression Profiling , Transcriptome , Antigens, Surface/genetics , Antigens, Surface/metabolism , Cell Line, Transformed , Cell Proliferation , Cells, Cultured , Flow Cytometry , Gene Expression , Genes, Viral , Genotype , /genetics , Leukocytes, Mononuclear/metabolism , Leukocytes, Mononuclear/virology , Transcription, Genetic , Virus Latency
Ciênc. saúde coletiva ; 20(4): 1099-1107, 04/2015. graf
Article in Portuguese | LILACS | ID: lil-744885


Trata-se de um estudo sobre o uso do ensino a distância (EaD) como uma estratégia de ensino na educação permanente em saúde (EPS), que teve como objetivo identificar e analisar os limites e possibilidades do uso da EaD na EPS. Estudo de revisão integrativa. O resultado aponta que a EaD é uma estratégia inovadora possível e potencial para a EPS, facilitando o desenvolvimento da aprendizagem dentro ou fora da instituição de saúde, porém é evidente a escassez de pesquisas na área. As limitações para a realização dos programas estão relacionadas à variável tempo, preparação para lidar com as tecnologias e importância do tutor como facilitador da aprendizagem. Conclui-se que o uso da EaD tem tido uma importante contribuição para o desenvolvimento dos recursos humanos em saúde, seja no processo de formação e/ou no processo contínuo de conhecimento.

This is a study on the use of distance learning (EaD, in Portuguese) as a teaching strategy in continuing health education (EPS, in Portuguese), which aimed to identify and analyze the limits and posibilities of using EaD in the EPS. Integrative Review Study. The result shows that EaD is an innovative, possible and potential strategy for EPS, facilitating the development of learning within or outside the health institution, although is evident the lack of research in the area. The limitations for the implementation of the programs are related to the time variable, preparation for dealing with the technologies and the importance of the tutor as a facilitator of learning. It concludes that the use of EaD has an important contribution to the development of human resources in health, is in the process of training and/or in the continuous knowledge process.

Female , Humans , Cell Proliferation , Gene Expression Regulation, Neoplastic , MicroRNAs/genetics , African Continental Ancestry Group/genetics , Basic Helix-Loop-Helix Transcription Factors/genetics , Cell Line, Transformed , Cell Line, Tumor , Cation Transport Proteins/genetics , Ethnic Groups , Europe , European Continental Ancestry Group/genetics , Genome-Wide Association Study , HapMap Project , Mitochondrial Proteins/genetics , Nigeria , Ovarian Neoplasms/genetics , Phenotype , Regression Analysis , Tumor Suppressor Proteins/genetics
Rev. méd. Chile ; 143(2): 237-243, feb. 2015. ilus
Article in Spanish | LILACS | ID: lil-742575


Currently, there is no discussion on the need to improve and strengthen the institutional health care modality of FONASA (MAI), the health care system used by the public services net and by most of the population, despite the widely known and long lasting problems such as waiting lists, hospital debt with suppliers, lack of specialists and increasing services purchase transference to the private sector, etc. In a dichotomous sectorial context, such as the one of health’s social security in Chile (the state on one side and the market on the other), points of view are polarized and stances tend to seek refuge within themselves. As a consequence, to protect the public solution is commonly associated with protecting the “status quo”, creating an environment that is reluctant to change. The author proposes a solution based on three basic core ideas, which, if proven effective, can strengthen each other if combined properly. These are: network financing management, governance of health care services in MAI and investments and human resources in networked self-managed institutions. The proposal of these core ideas was done introducing a reality testing that minimizes the politic complexity of their implementation.

Animals , Humans , Rats , AMP-Activated Protein Kinases/metabolism , Antioxidants/therapeutic use , Autophagy/drug effects , Signal Transduction/drug effects , Sirtuin 1/metabolism , Stilbenes/therapeutic use , Cell Line, Transformed , Dose-Response Relationship, Drug , Doxycycline/pharmacology , Gene Expression Regulation/drug effects , Insecticides/toxicity , Microscopy, Immunoelectron/methods , Microtubule-Associated Proteins/genetics , Microtubule-Associated Proteins/metabolism , Mutation/genetics , Poly(ADP-ribose) Polymerases/metabolism , RNA, Small Interfering/pharmacology , Rotenone/toxicity , Time Factors , alpha-Synuclein/genetics , alpha-Synuclein/metabolism
Article in English | WPRIM | ID: wpr-290169


Cementum is critical for anchoring the insertion of periodontal ligament fibers to the tooth root. Several aspects of cementogenesis remain unclear, including differences between acellular cementum and cellular cementum, and between cementum and bone. Biomineralization is regulated by the ratio of inorganic phosphate (Pi) to mineral inhibitor pyrophosphate (PPi), where local Pi and PPi concentrations are controlled by phosphatases including tissue-nonspecific alkaline phosphatase (TNAP) and ectonucleotide pyrophosphatase/phosphodiesterase 1 (NPP1). The focus of this study was to define the roles of these phosphatases in cementogenesis. TNAP was associated with earliest cementoblasts near forming acellular and cellular cementum. With loss of TNAP in the Alpl null mouse, acellular cementum was inhibited, while cellular cementum production increased, albeit as hypomineralized cementoid. In contrast, NPP1 was detected in cementoblasts after acellular cementum formation, and at low levels around cellular cementum. Loss of NPP1 in the Enpp1 null mouse increased acellular cementum, with little effect on cellular cementum. Developmental patterns were recapitulated in a mouse model for acellular cementum regeneration, with early TNAP expression and later NPP1 expression. In vitro, cementoblasts expressed Alpl gene/protein early, whereas Enpp1 gene/protein expression was significantly induced only under mineralization conditions. These patterns were confirmed in human teeth, including widespread TNAP, and NPP1 restricted to cementoblasts lining acellular cementum. These studies suggest that early TNAP expression creates a low PPi environment promoting acellular cementum initiation, while later NPP1 expression increases PPi, restricting acellular cementum apposition. Alterations in PPi have little effect on cellular cementum formation, though matrix mineralization is affected.

Alkaline Phosphatase , Metabolism , Animals , Cell Line, Transformed , Dental Cementum , Cell Biology , Metabolism , Physiology , Gene Expression Profiling , Humans , Mice , Models, Animal , Phosphoric Diester Hydrolases , Metabolism , Pyrophosphatases , Metabolism , Tooth Root , Metabolism , Physiology , X-Ray Microtomography
Article in English | WPRIM | ID: wpr-250360


Liver fibrosis is an important health problem that can further progress into cirrhosis or liver cancer, and result in significant morbidity and mortality. Inhibiting proliferation and inducing apoptosis of hepatic stellate cells (HSCs) may be the key point to reverse liver fibrosis. At present, anti-fibrosis drugs are rare. Kinetin is a type of plant-derived cytokinin which has been reported to control differentiation and induce apoptosis of human cells. In this study, the HSCs were incubated with different concentrations of kinetin. The proliferation of rat HSCs was measured by MTT assay, cell cycle and apoptosis were analyzed by flow cytometry, and the apoptosis was examined by TUNEL method. The expression of Bcl-2 and Bax proteins was detected by immunocytochemistry staining. It was found that kinetin could markedly inhibit proliferation of HSCs. In a concentration range of 2 to 8 μg/mL, the inhibitory effects of kinetin on proliferation of HSCs were increased with the increased concentration and the extension of time (P < 0.01). Flow cytometry indicated that kinetin could inhibit the DNA synthesis from G0/G1 to S phase in a dose-dependent manner (P < 0.01). The apoptosis rates of the HSCs treated with 8, 4 and 2 μg/mL kinetin (25.62% ± 2.21%, 15.31% ± 1.9% and 6.18% ± 1.23%, respectively) were increased significantly compared with the control group (3.81% ± 0.93%) (P < 0.01). All the DNA frequency histogram in kinetin-treated groups showed obvious hypodiploid peak (sub-G1 peak), and with the increase of kinetin concentrations, the apoptosis rate of HSCs also showed a trend of increase. It was also found that kinetin could down-regulate the expression of Bcl-2, and up-regulate the expression of Bax, leading to the decreased ratio of Bcl-2/Bax significantly. The kinetin-induced apoptosis of HSCs was positively correlated with the expression of Bax, and negatively with the expression of Bcl-2. It was concluded that kinetin can inhibit activation and proliferation of HSCs by interrupting the cell cycle at G1/S restriction point and inducing apoptosis of HSCs via reducing the ratio of Bcl-2/Bax.

Animals , Apoptosis , Cell Line, Transformed , Cell Proliferation , Dose-Response Relationship, Drug , G1 Phase Cell Cycle Checkpoints , Genetics , Gene Expression Regulation , Growth Inhibitors , Pharmacology , Hepatic Stellate Cells , Cell Biology , Metabolism , Kinetin , Pharmacology , Proto-Oncogene Proteins c-bcl-2 , Genetics , Metabolism , Rats , Signal Transduction , bcl-2-Associated X Protein , Genetics , Metabolism
Salud colect ; 10(3): 353-363, sep.-dic. 2014.
Article in Spanish | LILACS | ID: lil-733295


El objetivo de este artículo es analizar las relaciones entre profesionales de la salud y usuarios/as desde la perspectiva de género. A partir del referencial teórico de Pierre Bourdieu, se retoman y analizan críticamente datos de dos investigaciones realizadas en Brasil de la cuales participamos como autores: la primera, realizada en la ciudad de Río de Janeiro y, la segunda, un estudio multicéntrico y de naturaleza etnográfica desarrollado en ocho servicios distribuidos en cuatro estados brasileños, dos de la región Sudeste y dos de la Nordeste. Entre los principales resultados se destaca que las relaciones entre profesionales de la salud y usuarios/as, aunque estén marcadas por opiniones divergentes, están atravesadas por un habitus "generificado". Se concluye, entre otros aspectos, que la construcción de las diversas feminidades y masculinidades y la forma en que se da el ejercicio de estas en los contextos asistenciales de la salud son producto de un proceso que es, al mismo tiempo, socio-histórico y personal.

The objective of this article is to analyze relationships between health professionals and users from a gender perspective. Using Pierre Bourdieu as a theoretical reference, we critically analyze data from two studies carried out in Brazil in which we took part as authors. The first of these studies was based in Rio de Janeiro and the second was a multicenter and ethnographical study carried out in eight health care facilities distributed throughout four Brazilian states, two in the Southeast region and two in the Northeast region. Among the principal results of the present study, we found that although the relationships between health professionals and users demonstrate varied opinions, all are marked by a gendered habitus. We conclude that, among other aspects, the construction of diverse femininities and masculinities and the way in which these are exercised in health care contexts are the product of process that is both socio-historical and personal.

Humans , Esophageal Neoplasms/chemistry , Neoplasm Proteins/analysis , Cell Line, Transformed , Electrophoresis, Gel, Two-Dimensional , Gene Expression Regulation, Neoplastic , Neoplasm Proteins/genetics , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
Journal of Experimental Hematology ; (6): 1597-1601, 2013.
Article in Chinese | WPRIM | ID: wpr-264969


This study was aimed to explore the method for induction and expansion of EB virus specific cytotoxic T lymphocytes (EBV-CTL) in vitro, and to detect their killing effect. Peripheral blood mononuclear cells (PBMNC) were collected from 6 EBV seropositive healthy donors, and EBV-transformed B lymphoblastoid cells (BLCL)were used as the antigen-presenting cells and antigen stimulant which was irradiated by 40 Gy (60)Co irradiator. The autologous PBMNC and irradiated BLCL were cultured to induce and expand the EBV-CTL, and the immunophenotype was identified by the flow cytometry. The killing effect of the EBV-CTL against the autologous BLCL (autoBLCL), the autologous PHA cultured B lymphoblastoid cells( PHA-BLCL), the allogeneic BLCL (alloBLCL) and the K562 cells were measured with LDH release assay under different effector-to-target ratio. The results showed that the 6 cell lines of EBV-CTL were induced and expanded from the EBV seropositive healthy donors, the overall increase in cell numbers varied from 18.6 to 55.0 times. After 10 stimulations, the specific killing efficiency of the EBV-CTL for the autoBLCL were 59.4%, 43.2% and 29.0% under the effector-to-target ratio of 20: 1, 10: 1 and 5: 1. The nonspecific killing efficiency for the PHA-blast, alloBLCL and K562 cells were 7.1%, 9.4% and 10.3% (P < 0.05) under the 20: 1 ratio; 6.6%, 8.3% and 8.1% (P < 0.05) under 10: 1; 5.4%, 7.3% and 6.3% (P < 0.05) under 5: 1, respectively. It is concluded that the EBV-CTL can be successfully induced and expanded ex vivo for specific killing of HLA matched BLCL and may become a potential treatment for EBV related post-transplant lymphoproliferative disorders.

B-Lymphocytes , Allergy and Immunology , Cell Line, Transformed , Herpesvirus 4, Human , Allergy and Immunology , Humans , K562 Cells , Leukocytes, Mononuclear , Allergy and Immunology , Virology , T-Lymphocytes, Cytotoxic , Cell Biology , Allergy and Immunology , Virology
Article in English | IMSEAR | ID: sea-140327


Background & objectives: A major drawback for genetic studies as well as long-term genotype-phenotype correlation studies in cancer is lack of representative human cell lines providing a continuous source of basic biomolecules and a system to carry out various experimental investigations. This can be overcome to some extent by establishing lymphoblastoid cell lines (LCLs) by infecting peripheral blood lymphocytes with Epstein Barr virus (EBV) which is known to immortalize human resting B cells in vitro giving rise to actively proliferating B-lymphoblastoid cell lines. The present study involves preparation and characterization of LCLs generated from patients with multiple primary neoplasms (MPN) of upper aero-digestive tract (UADT). Methods: Thirty seven LCLs were established from UADT MPN patients and healthy age, sex and habit matched controls using EBV crude stock. Characterization was done with respect to expression of CD-19 (Pan B-cell marker), CD3 (T cell specific marker), CD56 (NK-cell specific marker), cell morphology, ploidy analysis, genotype and gene expression comparison with the parent lymphocytes. Results: LCLs showed rosette morphology with doubling time of approximately 24 h. Ploidy analysis showed diploid DNA content which was maintained for at least 30 population doublings. When compared with parent lymphocytes there appeared no change at genetic and gene expression level. Interpretation & conclusions: Our results show that lymphoblastoid cell lines are a good surrogate of isolated lymphocytes bearing their close resemblance at genetic and phenotypic level to parent lymphocytes and are a valuable resource for understanding genotype-phenotype interactions.

Herpesvirus 4, Human/analysis , Herpesvirus 4, Human/isolation & purification , Humans , Cell Line, Transformed , Cell Line , Neoplasms, Multiple Primary , Patients , Ploidies
Braz. j. biol ; 71(2): 487-490, maio 2011. ilus, tab
Article in English | LILACS | ID: lil-592585


Treatment of human breast epithelial cells MCF-10F with 17-β-estradiol has been reported to result in E2-transformed cells which have given rise to highly invasive C5 cells that in turn generate tumors in SCID mice. From these tumors, various cell lines, among which C5-A6-T6 and C5-A8-T8, were obtained. Although different phases of the tumorigenesis process in this model have been studied in molecular biology and image analysis assays, no cytological data on apoptotic ratios and mitotic abnormalities have been established to accompany the various steps leading to 17-β-estradiol-treated MCF-10F cells to tumorigenesis. Here we detected that the apoptotic ratio decreases with the transformation and tumorigenesis progress, except for the tumor cell line C5-A8-T8, probably on account of its more intense proliferation rate and a more rapid culture medium consumption. Increased frequency of mitotic abnormalities contributed by triple- and tetrapolar metaphases, and by lagging chromosomes and chromosome bridges observed at the anaphase found by transformation and tumorigenesis progress. However, no difference was found under these terms when the C5-A6-T6 and C5-A8-T8 tumor cell lines were compared to each other. Present findings are in agreement with the nuclear instability and enrichment of dysregulated genes in the apoptotic process promoted by transformation and tumorigenesis in 17-β-estradiol-treated MCF-10F cells.

O tratamento das células epiteliais mamárias humanas MCF-10F com 17-β-estradiol tem sido relatado como resultando nas células transformadas E2, que deram origem às células C5, altamente invasivas, e que geraram tumores em camundongos SCID. A partir desses tumores foram originadas em cultura células tumorais, dentre as quais C5-A6-T6 e C5-A8-T8. Embora diversas fases do processo tumorigênico neste modelo tenham sido estudadas por ensaios de biologia molecular e análise de imagem, não foram ainda estimados dados citológicos referentes a índices apoptóticos e anomalias mitóticas que acompanhassem os vários passos que levam as células CF-10F tratadas com 17-β-estradiol à tumorigênese. Neste trabalho detectamos que o índice apoptótico decresce com a transformação e o avanço da tumorigênese, exceto na linhagem celular tumoral C5-A8-T8, provavelmente por causa de sua velocidade de proliferação mais intensa, que poderia levá-la a um consumo mais rápido do meio de cultura presente e à morte celular. Um aumento na frequência de anomalias mitóticas contribuídas por metáfases tripolares e tetrapolares e por pontes cromossômicas e cromossomos desgarrados, identificáveis na anáfase, foi observado com a transformação e o progresso da tumorigênese. Contudo não foram detectadas diferenças nesses parâmetros quando se compararam as linhagens tumorais C5-A6-T6 e C5-A8-T8 entre si. Os presentes achados estão de acordo com a instabilidade nuclear e o enriquecimento em desregulação de genes que atuam no processo apoptótico, promovidos pela transformação e tumorigênese nas células MCF-10F tratadas com 17-β-estradiol.

Animals , Female , Humans , Mice , Apoptosis/drug effects , Breast Neoplasms/pathology , Epithelial Cells/drug effects , Epithelial Cells/pathology , Estradiol/pharmacology , Mitosis/drug effects , Cell Line, Transformed , Cell Line, Tumor , Cell Transformation, Neoplastic
Chinese Journal of Virology ; (6): 515-520, 2011.
Article in Chinese | WPRIM | ID: wpr-354797


Severe fever with thrombocytopenia syndrome bunyavirus (SFTSV) is a novel phlebovirus, causing a life-threatening illness associated with the symptoms of severe fever and thrombocytopenia syndrome. The sequence and structure of the genome have already been illustrated in previous study. However, the characteristics and function of the structure and non-structure proteins is still unclear. In this study, we identified the density of the purified SFTSV virions as 1.135 g/mL in sucrose solution. Using RT-PCR method, we amplified the full coding sequence of RNA dependent RNA polymerase(RdRp), glycoprotein precursor (M), glycoprotein n (Gn), glycoprotein c (Gc), nuclear protein (NP) and non structural protein (NSs) of SFTSV (strain HB29). Respectively inserted the target genes into eukaryotic expression vector pcDNA5/FRT or VR1012, the target protein in 293T cell were successfully expressed. By analyzing the SFTSV virions in SDS-PAGE and using recombinant viral proteins with SFTS patients sera in Western blotting and Immunofluorescent assay, the molecule weight of structure and non-structure proteins of SFTSV were defined. The study provides the first step to understand the molecular characteristics of SFTSV.

Bunyaviridae Infections , Virology , Cell Line, Transformed , Fever , Virology , HEK293 Cells , Humans , Orthobunyavirus , Genetics , Metabolism , Thrombocytopenia , Virology , Viral Nonstructural Proteins , Genetics , Viral Structural Proteins , Genetics , Virion , Genetics , Metabolism
Chinese Journal of Virology ; (6): 587-593, 2011.
Article in Chinese | WPRIM | ID: wpr-354787


To develop a stable cell line that could express the RSV NS1, the full-length RSV NS1 gene was generated by RT-PCR amplification from respiratory syncytial virus. NS1 gene was ligated with pBABE-puro to construct the recombinant retroviral expression plasmid pBABE-NS1, which was cotransfected into 293FT packaging cells with PIK packaging plasmid by calcium phosphate co-precipitation. The supernatant of 293FT was collected to infect HEp-2 cells, the resulting cell clones stably expressing NS1 were screened by puromycin. Using QPCR, CPE staining method and indirect immunofluorescence assay, the expression of NS1 at both gene and protein levels was identified. The recombinant plasmid pBABE-NS1 was identified by EcoRI and BamHI endonuclease digestion and the sequence analysis. QPCR results showed that the NS1 gene amplification in HEp-2-NS1 cells was 8483 fold higher than that in HEp-2 cells. Although the exogenous interferon was added, all cells were destroyed after 48 hours post infection using CPE staining method, showing that HEp-2-NS1 cells remained sensitive to the VSV virus. The results of RT-PCR and indirect immunofluorescence assay showed that the NS1 gene in HEp-2 cells could not only transcribe mRNA, but also express NS1 protein steadily. We had successfully established HEp-2-NS1 cell lines with stable expression of respiratory syncytial virus non-structural protein NS1.

Cell Line, Transformed , HEK293 Cells , Humans , Recombinant Proteins , Genetics , Respiratory Syncytial Viruses , Genetics , Viral Nonstructural Proteins , Genetics