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Item Modulation of heat shock protein expression and cytokine levels in MCF‐7 cells through photodynamic therapy(Springer-Verlag London Ltd.) Santos, Mariela Inês Batista dos; Godoi, Bruno Henrique; Silva, Newton Soares da; Oliveira, Luciane Dias de; Ramos, Lucas de Paula; Cintra, Ricardo Cesar; Pacheco‐Soares, CristinaIn this study, we assess the impact of photodynamic therapy (PDT) using aluminum phthalocyanine tetrasulfonate (AlPcS4) on the viability and cellular stress responses of MCF-7 breast cancer cells. Specifically, we investigate changes in cell viability, cytokine production, and the expression of stress-related genes. Experimental groups included control cells, those treated with AlPcS4 only, light-emitting diode (LED) only, and combined PDT. To evaluate these effects on cell viability, cytokine production, and the expression of stress-related genes, techniques such as 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) assay, enzyme-linked immunosorbent assays (ELISA), and real-time quantitative PCR (RT‒qPCR) were employed. Our findings reveal how PDT with AlPcS4 modulates mitochondrial activity and cytokine responses, shedding light on the cellular pathways essential for cell survival and stress adaptation. This work enhances our understanding of PDT's therapeutic potential and mechanisms in treating breast cancer.Item Estresse oxidativo em células SH-SY5Y diante da exposição ao peróxido de hidrogênio(CDRR Editors) Zabeu, Antonieta Marques Caldeira; Rossato, Rafaella Carvalho; Pacheco-Soares, Cristina; Silva, Newton Soares daO estresse oxidativo é consequência da formação excessiva de espécies reativas de oxigênio geradas normalmente pelo metabolismo celular. Essa produção excessiva ocasiona a perda de capacidade de defesa e de reparo, levando a danos nas biomoléculas (DNA, lipídios, proteínas) celulares. Quando estes danos não são reparados, acabam comprometendo o funcionamento da célula, levando-a a morte por apoptose ou necrose. O estresse oxidativo está associado à morte de células neurais como sendo uma das principais causas de doenças neurodegenerativas, incluindo doença de Alzheimer e doença de Parkinson. O objetivo deste estudo é verificar a ação do peróxido de hidrogênio sendo um modelo de estresse oxidativo em células de neuroblastoma humano. Busca-se analisar a partir de qual concentração ocorre um estresse significante nas células e, em conjunto, verificar esse limiar de concentração em diferentes tempos de ação nas células. Os experimentos realizados compreenderam as análises de atividade mitocondrial obtido por meio do teste de corante tetrazólio (MTT), teste de viabilidade celular utilizando o ensaio de azul de tripano, obtendo-se assim dados quantitativos e qualitativos. Foi possível observar que, a partir da concentração de 0,2 mM durante um intervalo de tempo de 2 horas houve uma diminuição significativa no número de células viáveis em relação ao grupo controle. E, acima dessa concentração, em relação ao tempo, ocorreram mais danos celulares.Item Photobiomodulation assay of muscle cells C2C12 after irradiation with LED device(CDRR Editors) Lima, Elessandro Váguino de; Pacheco-Soares, Cristina; Silva, Newton Soares daIntroduction:One of the ways that have been observed to reduce musculoskeletal fatigue is the use of protocols for the application of light sources (photobiomodulation) such as low-intensity laser and LED (Light Emitting Diode). Work involving photobiomodulation has shown promising results in strength performance or reduction of muscle fatigue. At the cellular level,photobiomodulation can modulate fibroblasts proliferation, the fixation and synthesis of collagen and procollagen, promote angiogenesis and improving energy metabolism in mitochondria. Compared with laser devices, LED has several advantages, such as beingsmaller, lighter, lower cost, and easier for operation. Objective:The present work objective is to verify if irradiation with LED device (650 nm and 860 nm) in muscle cells C2C12 modify the viability, morphology and cytoskeleton components. Methodology:C2C12 cells line (ATCC CRL -1772) were cultured in 25 cm2bottles at 37ºC under 5% CO2in DMEM. The cells wereirradiated with the light-emitting diodes (LED) device, Sportllux Ultra that consists of 84 LEDs, each individual LED has 8 mW of power, emitting in 660±20 nm (42 LEDs) and 850±20 nm (42 LEDs), and covering anarea (A) of 120 cm2. The power density of delivered light was 5,6 mW/cm2, and the exposure time was 10 minutes, totalizing the fluence of 3,4 J/cm2. Viability assay was performed where the cells were incubated with 100 μL of Crystal Violet (CV) solution and mitochondrial activity assay was evaluated by the colorimetric MTT assay. Nucleus (DAPI) and Cytoskeleton (Rhodamine Phalloidin) fluorescence assay was performed to study the cytoskeleton based on the change in the actin filaments. Results:Our results demonstrate that the synergism of LED irradiation (660nm and 850nm) induced the proliferation of C2C12 cells. The light-emitting diode (LED) device, Sportllux Ultra has a significant effect on C2C12 cell culture. Mitochondrial activity and cell viability showed a significative increase in their activities after irradiation. The microscopy fluorescence observations showed an alignment of cytoskeletal components of C2C12 cells after irradiation.Conclusion:The application of irradiation with the Sportlux Ultra LED device stimulated an increase of energy by mitochondrial activity assay, number of cells by cell viability assay and alignment of cytoskeleton components by fluorescence assay in C2C12line cells. Our results suggest that organizated cytoskeletal actin filaments normally contribute to cell survival and that induced major cell changes in the cytoskeleton that result in cell shape change. These results suggest that the Sport Lux Ultra LEDdevice can help in the repair of tissue injuries and to collaborate to increase of performance in athletes in a faster wayItem Performance of Multiplex Detection Method of IgM Class Antibodies against Toxoplasma gondii, Rubella and Human Cytomegalovirus(JSciMed Central) Almeida, Rainara Moreno Sanches de; Ogawa, Guilherme Maerschner; Custódio, Paulo Rogério Siqueira; Sampaio, Maria Alcantara Socorro; Silva, Newton Soares da; Soares, Cristina PachecoSerological diagnosis during neonatal screening is crucial in disease prevention. Among the infectious diseases, the most common are toxoplasmosis, rubella, and cytomegalovirus. Traditional diagnostic methods are used to detect a single infectious agent per test. The use of multiplex detection methods increases productivity and reduces the amount of material used, resulting in a more efficient test from a technical, environmental, and economic point of view. The study’s objective was to evaluate the performance of a new diagnostic method aimed at neonatal screening using the multiplex platform of magnetic microspheres from the company Luminex Corporation. For this, tests were carried out for analytical validation of the diagnostic product developed following the rules of the National Health Surveillance Agency (ANVISA) of Brazil. The parameters evaluated were repeatability, reproducibility, linearity, robustness, high dose, minimum detection limit, and analytical specificity. All data obtained met the acceptance criteria of RDC 166/17 of 2017 for the use of the diagnostic product in the national territory. Repeatability and reproducibility tests showed a CV of less than 15% between replicates of the same operator and different operators. The kit showed linearity throughout the operating range with R2 above 0.990, and no effect of high-efficiency dose was observed in the chosen working dilution. In addition, the kit did not show interference from the matrix with the results, and it was observed that small and deliberate changes in the incubation time of each reagent did not have a significant effect on the data obtained.Item Evaluation of the effect of hydrocortisone in 2D and 3D HEp-2 cell culture(CDRR Editors) Fonseca, Marcelo de Oliveira; Godoi, Bruno Henrique; Silva, Newton Soares da; Pacheco-Soares, CristinaIntroduction: Cancer is one of the diseases with the highest incidence globally and that associated with the patient'semotional state, can act positively or negatively in the treatment. Cortisol is a principal primary stress hormone in the human body. The corticoids can increase cell proliferation and reactive oxygen species that contribute to DNA damage. Prolonged exposure to stress can contribute to tissues becoming insensitive to cortisol, the primary human stress hormone. Objective: This study explores cortisol's influence on tumor cell development, particularly in human cells of carcinoma of the human laryngeal (HEp-2). Methodology: HEp-2 cells were exposed toincreasing cortisol (hydrocortisone) concentrations for 24 or 48 hours, and cytotoxicity (MTT assay) proliferation assay (crystal violet assay), and immunolabeled 3D culture for fibronectin and FAK were analyzed. Results: The group treated with hydrocortisone showed a significant increase in mitochondrial activity, as for the evaluation by the violet crystal, the treated group showed similar behavior to the control. The 3D culture showed dispersed cells within 24hours with reduced FAK labeling; however, no changes were observed within 48 hours. Conclusion: Although some cases favored corticosteroid use in cancer patients, a more detailed analysisis necessary before prescribing them.Item Influence of Hydrocortisone in Chemotherapy and Photodynamic Therapy in HEp-2 Cells(Clinics in Oncology) Moraes, Carlos Dailton Guedes de Oliveira; Godoi, Bruno Henrique; Silva, Newton Soares da; Soares, Cristina PachecoAim: Cancer cells exhibit resistance to the immune response by regulating and altering the expression of mediators responsible for immune cell recruitment and disease progression. Cortisol is a natural hormone that may be associated with diseases such as cancer by stimulating stress and altering the cellular environment, favoring uncontrolled division and contributing to the inhibition of the immune response. In contrast, current therapeutic strategies do not present significant concerns about stress as a variable in cancer diagnosis and prognosis. The response of HEp-2 cells to stress induced by hydrocortisone and to treatment with Cyclophosphamide (CP) and Photodynamic Therapy (PDT) was analyzed. Methods: One mM of hydrocortisone induced stress in the cells. Cells were treated with 200 μg/ mL of cyclophosphamide or Aluminum Phthalocyanine Tetrasulfonate (AlPcS4) photosensitizer, LED irradiation (660 nm wavelength), intensity of 25 mW/cm2, power of 70 mW, fluence of 5 J/ cm2, characterizing the PDT. All groups were evaluated after 24 h and 48 h. Results: Assessment of stress-inducing mitochondrial activity and cell viability were performed, and the results demonstrated that hydrocortisone significantly altered the rate of cell death, compromising the effects of CP. Conclusion: However, hydrocortisone did not change the cell death rates caused by PDT, indicating the possibility of this hormone as an alternative therapy.Item Anatase film on orotracheal tubes to mitigate Staphylococcus aureus(American Scientific Publishers) Manfroi, Lucas Augusto; Silva, Michely Glenda Pereira da; Vieira, Angela Aparecida; Macário, Paulo Fabrício; Silva, Newton Soares da; Marques, Francisco Chagas; Vieira, LuciaBacterial contamination in hospital environments is a significant concern for patient admissions. Aiming to reduce contamination, titanium dioxide film (TiO2) in the anatase phase has been prepared on the surface of polyvinyl chloride (PVC) tubes. The PVC tube material was used to study the film’s effectiveness in inhibit- ing bacterial growth and cell viability. The morphology and composition of deposited films were investigated using a Scanning Electron Microscope (SEM) and Energy Dispersive Spectroscopy (EDS) map. In addition, Fourier-Transform Infrared Spectroscopy (FTIR) and XRD diffractogram were used to analyze film composition and phase, respectively. The adhesion of TiO2 film on PVC substrate was determined using ScotchTM tape-test according to ASTM: D3359-09, 2010, and the film surface morphology was analyzed by the MEV-FEG tech- nique and EDS map. The bacterial viability was performed with Staphylococcus aureus, and cell viability was performed using L929 strain mouse fibroblasts. The results of TiO2 in the anatase phase deposited by ALD on the PVC surface demonstrate good adherence and the film’s effectiveness in inhibiting bacterial growth and cell viability.Item Ação de extrato de folhas de Acmella oleracea (L.) R. K. Jansen em co-cultivo de Staphylococcus aureus e L929 (fibroblastos) simulando processo de celulite infecciosa(CDRR Editors) Silva, Laís Eduarda; Silva, Carlos Augusto Priante da; Pacheco-Soares, Cristina; Silva, Newton Soares daIntrodução: A bactéria Staphylococcus aureus é o agente etiológico de maior incidência nas infecções de pele, dentre elas a celulite infecciosa. Seu tratamento é contido por antibióticos, porém o uso de plantas medicinais vem sendo utilizadas para tratamento. Acmella oleracea é uma planta da família Asteraceae conhecida como jambu. Objetivos: Avaliar a ação do extrato das folhas de Acmella oleracea em co-cultivo da linhagem celular L929 e bactérias Staphylococcus aureus simulando o processo de celulite infecciosa. Metodologia: Foi realizado primeiramente o teste MTT e Cristal Violeta na linhagem celular L929 incubadas com o extrato do jambu, nas concentrações 500 µg/mL, 1000 µg/mL e 2000 µg/mL. Foi realizado ensaios com Staphylococcus aureus incubado no período de 24 horas com o extrato do jambu nas mesmas concentrações. Resultados: Na avaliação das células L929 no teste MTT houve uma significância na concentração de 1000 µg/mL apresentando uma baixa atividade metabólica em relação as outras concentrações. No teste cristal violeta ocorreu um maior estímulo nas demais concentrações quando comparado ao controle. Na viabilidade da bactéria observou-se uma queda significativa da viabilidade quando comparado ao controle. A concentração de 500 ug/mL foi que apresentou um menor índice de UFC. Ao realizar o teste do co-cultivo L929 - S. aureus após a incubação de 24 horas com diferentes concentrações do extrato, foi possível avaliar que a ação do extrato apresentou uma redução significativa na viabilidade bacteriana e não interferência à viabilidade das células L929. Conclusão: Com estes resultados demonstramos que o extrato de Acmella oleracea apresentou uma atividade antimicrobiana sem interferir na linhagem de fibroblastos L929, sendo assim podendo ser utilizada contra a celulite infecciosa.