9 resultados
Resultados de Busca
Agora exibindo 1 - 9 de 9
Item 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 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 Estimation of the DNA damage in Girardia tigrina (Girard, 1850) after exposure to radiation under extreme conditions(Brazilian Journals Publicações de Periódicos) Benitz, Tabatha; Oliveira, Matheus Salgado de; Lopes, Karla Andressa Ruiz; Soares, Cristina Pacheco; Morais, Flavia Villaça; Velho, Nádia Maria Rodrigues de CamposThe use of planarians as experimental models has been increased worldwide especially due to their high capacity of tissue regeneration and ease of laboratory maintenance. The current study aims to assess the genetic damage of Girardia tigrina after exposure of stressors in conditions such as: critical temperature, critical pH, hypergravity (4.4G) and low power laser (685nm). 18 specimens of G. tigrina were used, the extreme conditions mentioned were experimental group with 3 specimens each, and 3 for the control group, all specimens were submitted to the Comet Assay after exposure of stressors, and control group with no extreme condition exposure. The extreme stimuli of critical temperature and pH, hypergravity and low power laser irradiation, caused genetic damages to the G. tigrina, for hypergravity and low-power laser stressors the damage was higher in the pharyngeal and post-pharyngeal regions of the animal with a greatest area of comet tail dragging for the post-pharyngeal region. The extreme stimuli of critical temperature and pH, hypergravity and low power laser irradiation caused genetic damages in planarians Brazilian Journal of Development ISSN: 2525-8761 12160 Brazilian Journal of Development, Curitiba, v.8, n.2, p.12159-12177 feb. 2022. G. tigrina, especially in samples from the pharyngeal and post-pharyngeal regions of the flatworms body assessed by the Comet Assay.Item Biomodulatory effect of low intensity laser (830 nm.) in neural model 9L/lacZ(CDRR Editor) Zabeu, Antonieta Marques Caldeira; Carvalho, Isabel Chaves Silva; Soares, Cristina Pacheco; Silva, Newton Soares daCurrently, research is advancing with low-intensity laser (LIL) in cells of the central nervous system, with the aim of evaluating the benefits of this therapy in neurological disorders such as Alzheimer's, stroke, ischemia, epilepsy, among others. The aim of this study was to verify the biomodulatory and biostimulatory effects of LIL in neural cell culture. Diode laser at wavelength λ = 830 nm, power 40 mW, in continuous mode, was applied on the 9L/lacZ cell line with energy densities of 0.5 to 3 J/cm . The analysis was performed 24 hours after irradiation, the results of cell viability showed a difference between the control and irradiated groups. As for the occurrence of apoptosis, no significant manifestation was observed between the control group compared to the irradiated group (P = 0.9956); there was a significant difference between apoptosis and death by necrosis between the control and treated groups (P<0.001). In the comet assay no statistically significant differences were observed. Regarding the objective of evaluating whether LIL promotes early activation of apoptosis or proliferation of 9L/lacZ cells at different energy densities of the infrared diode laser, we observed an increase in the number of neural cells, highlighting the action of biomodulation. Furthermore, LIL did not promote the activation of programmed cell death - apoptosis and did not show any indication of DNA damage by the comet assay. The results of this study are indicative that the laser in the near infrared has a positive interaction with neuronal cells.Item Neuroprotective effects of taurine on SH-SY5Y cells under hydrocortisone induced stress(CDRR Editors) Rossato, Rafaella Carvalho; Granato, Alessandro Eustaquio Campos; Moraes, Carlos Dailton Guedes de Oliveira; Salles, Geisa Nogueira; Soares, Cristina PachecoAlzheimer's disease (AD) is the most common, progressive and irreversible neurodegenerative disorder, characterized by memory loss, cognitive impairment and behavioral abnormalities. Although there is no cure, several study strategies seek to elucidate mechanisms of the disease. Recent studies address the benefits of taurine. Thus, the present study aims to analyze neuroprotective effects of taurine in human neuroblastoma (SH-SY5Y), using an in vitro experimental model of oxidative stress induced by hydrocortisone. This work showed for the first time that taurine can promote neuroprotection in SH-SY5Y under oxidative stress caused by hydrocortisone. Cell viability was evaluated using crystal violet and the evaluation of cell morphology was performed by scanning electron microscopy (SEM). The viability of SH-SY5Y pre-treated with taurine and stressed with hydrocortisone was preserved, compared to the stressed only group, which was also morphologically observed. Therefore, taurine can represent a considerable therapeutic candidate in the prevention of neurodegenerative diseases, such as AD.Item Juglans regia and Pfaffia paniculata extracts: implications for periodontal disease treatment and correlation with Alzheimer's risk(Frontiers) Miranda, Diego Garcia; Carrouel, Florence; Attik, Nina; Araujo, Gabriela Ferraz; Lopes, Nicole Fernanda dos Santos; Marcucci, Maria Cristina; Rodrigues, Flavia Pires; Caires, Giovanna Arruda; Vigerelli, Hugo; Godoi, Bruno Henrique; Soares, Cristina Pacheco; Ramos, Lucas de PaulaPeriodontal disease (PD) is a significant global health concern, affecting approximately 19% of the world’s population. It is one of the most prevalent diseases today, causing substantial socio-economic impacts and diminished quality of life. Recent research has also revealed a potential link between PD and Alzheimer’s disease. This study investigated the antimicrobial effects of Juglans regia and Pfaffia paniculata extracts against P. endodontalis and P. gingivalis, bacteria that cause PD and are related to Alzheimer’s risk. The study also assessed the impact of these extracts on macrophage metabolic activity, pro- and anti-inflammatory cytokine expression, and genotoxicity. The phytochemical analysis of the extract was carried out first. Antimicrobial activity was performed using the M11-A7 protocol (CLSI) for planktonic cultures on monotypic biofilms matured for 168 hours in anaerobiosis. Cell viability analysis was carried out using MTT on mouse macrophages (RAW 264-7), as well as genotoxicity assessment using micronuclei. The anti-inflammatory activity was evaluated using ELISA method, checking the cytokines IL-6, IL-1B, TNF-alpha, IL-17 and IL-10. Phytochemical analysis revealed the presence of Miquelianin, Regiolone and Gallic Acid in J. regia extract. For the P. paniculata extract, we identified the glycosides Pfaffoside C, Pfaffoside A, 3-O-β-D-glycopyranosyl-oleanolic acid and Beta-ecdysone. Antimicrobial activity revealed a MBC of 1.73 for the extract of J. regia and 0.48 for P. paniculata against P. endodontalis and P. gingivalis. All biofilms were reduced by more than 89% after treatment with the extracts for 5 min. Cytotoxicity evaluations revealed that cell viability remained above 50% at concentrations up to 0.216 mg/ml for J. regia and 0.015 mg/ml for P. paniculata. Neither extract exhibited genotoxicity. Furthermore, both demonstrated anti-inflammatory activity by promoting the production of the cytokine IL-10. In conclusion, the antimicrobial and anti-inflammatory activities of J. regia and P. paniculata extracts suggest their potential as treatments for oral dysbiosis, which may contribute to a reduced risk of neurodegenerative diseases.Item Gymnema sylvestre as a Potential Anti-Inflammatory and Anti-Biofilm Agent Against Anaerobic Infections: An In Vitro Study(MDPI) Miranda, Diego Garcia; Tome, Fernanda Malagutti; Miguel, Manuela Maria Viana; Liberato, Sabrina Ferreira dos Santos; Marcucci, Maria Cristina; Vigerelli, Hugo; Rodrigues, Flavia Pires; Soares, Cristina Pacheco; Godoi, Bruno Henrique; Carrouel, Florence; Oliveira, Luciane Dias de; Ramos, Lucas de PaulaThis study evaluates the antimicrobial activity of the glycolic extract of G. sylvestre against anaerobic pathogens, along with its cytotoxicity, genotoxicity, anti-inflammatory activity, antioxidant effects, and phytochemical composition. Phytochemical analysis was conducted using high-performance liquid chromatography and liquid chromatography– mass spectrometry, while the antioxidant effect was assessed through a DPPH assay. An- timicrobial action was tested on planktonic cultures and biofilms of Porphyromonas gingivalis, Porphyromonas endodontalis, Parvimonas micra, and Fusobacterium nucleatum. Cytotoxicity was evaluated using mouse macrophages (RAW 264.7), rat fibroblasts (L929), and human keratinocytes (HaCaT). Anti-inflammatory effects were measured by an immunoenzymatic assay (ELISA) on RAW 264.7 cells. Statistical analysis was performed using a one-way ANOVA and Tukey’s test. Phytochemical analysis revealed the presence of phenolic com- pounds and flavonoids. The extract demonstrated a reduction of over 95% in biofilms of P. gingivalis, P. micra, and F. nucleatum within 5 min of treatment. Cell viability (HaCaT) remained above 80%. Antioxidant activity showed an EC50 of 353.43 μg/mL, achieving a 50% reduction in free radicals. A significant decrease in TNF-α (a pro-inflammatory cytokine) and an increase in IL-10 (an anti-inflammatory cytokine) were observed. In conclusion, the extract of G. sylvestre exhibits promising potential as a therapeutic agent for treating anaerobic infections, inflammation, and oxidative stress.Item Photobiomodulation by LED 660 nm and Taurine against H2O2 oxidative stress in SH-SY5Y cells(Springer Nature Link) Rossato, Rafaella Carvalho; Salles, Geisa Rodrigues; Albuquerque, Amanda Lira; Porcionatto, Marimélia Aparecida; Granato, Alessandro Eustáquio Campos; Ulrich, Henning; Santos, Mariela Inês Batista dos; Soares, Cristina PachecoAlzheimer's Disease (AD) is a progressive uncurable neurodegenerative pathology affecting millions worldwide. Photo- biomodulation and Taurine are promising alternatives for preventing and reducing the rapid progression of neurodegenera- tion, stimulating the reconstructing of neural tissue structures, especially improving mitochondrial activity, which is highly impaired in AD. In this study, the mitochondrial effects of Taurine combined with light emitting diode (LED) irradiation were evaluated on human neuroblastoma cells (SH-SY5Y), under oxidative stress condition by hydrogen peroxide (H2O2) exposure, a considerable modulator in AD. We evaluated LED irradiation at the wavelength of 660 nm and Taurine under different concentrations before and together with exposing SH-SY5Y cells to different concentrations of H2O2, assessing mitochondrial activity by the MTT colorimetric test and labeling live cells mitochondria by the fluorescent probe MitoTracker. Cell viability was also evaluated by the trypan blue exclusion assay, and cellular morphological structures were imaged by scanning electron microscopy (SEM). Neuroprotective effects were achieved by both LED irradiation and LED irradia- tion + Taurine when cells were exposed to them before H2O2-induced stress. Comparing both agents, LED irradiation at 660 nm is sufficient to improve mitochondrial activity, however, healthy mitochondrial morphology was only observed when cells were treated with Taurine together with LED irradiation, representing affordable candidates that act in synergy against oxidative stress, one of the main contributors to neurodegeneration.Item Self-assembly and 3D Bioprinting of Neurospheres and Evaluation of Caffeine and Photobiomodulation Effects in an Alzheimer's Disease In Vitro Model(Springer Nature Link) Salles, Geisa Rodrigues; Granato, Alessandro Eustáquio Campos; Viero, Fernanda Tibolla; Soares, Cristina Pacheco; Ferreira, Sergio Teixeira; Porcionatto, Marimélia Aparecida; Ulrich, HenningSeveral in vitro models of Alzheimer’s disease (AD) rely on 2D cell culture, and, more recently, 3D cultures represented by free-floating neurospheres have been used as models for the disease. The advantage of 3D over 2D cell culture is that cell-extracellular matrix and cell-cell interactions can be assessed, better representing the molecular and cellular hallmarks of the disease. In the current study, we developed two complementary 3D neurosphere models using SH-SY5Y human neuroblastoma cells to investigate AD pathology and evaluate potential therapies. First, self-assembled neurospheres were exposed to hydrogen peroxide (H2O2) and amyloid-beta oligomers (AβOs), inducing AD-like features such as increased production of reactive oxygen species (ROS), amyloid aggregation, and apoptosis. Treatment with caffeine or photo- biomodulation (PBM) using LED irradiation significantly reduced Aβ1−42 accumulation, ROS generation, and decreased apoptosis markers. Second, 3D bioprinting of SH-SY5Y cells resulted in neurospheres with enhanced cellular organization and differentiation. These findings emphasize the advantages of 3D models for studying neurodegeneration and evaluating therapeutic strategies, bridging the gap between traditional 2D cultures and complex in vitro systems.