SUELY KAZUE NAGAHASHI MARIE
Projetos de Pesquisa
Unidades Organizacionais
Departamento de Neurologia, Faculdade de Medicina - Docente
LIM/15 - Laboratório de Investigação em Neurologia, Hospital das Clínicas, Faculdade de Medicina - Líder
LIM/15 - Laboratório de Investigação em Neurologia, Hospital das Clínicas, Faculdade de Medicina - Líder
9 resultados
Resultados de Busca
Agora exibindo 1 - 9 de 9
bookPart Sequenciamento em larga escala: impactos em oncologia(2017) OBA-SHINJO, Sueli Mieko; MARIE, Suely Kazue NagahashibookPart Biologia molecular das neoplasias do sistema nervoso central(2013) MARIE, Suely Kazue Nagahashi; SHINJO, Sueli Mieko ObabookPart Biologia Molecular em Neurologia Clínica(2015) MARIE, Suely Kazue Nagahashi; SHINJO, Sueli Mieko ObabookPart Dor Crônica(2015) GARCIA, Juliana Bueno; MARIE, Suely Kazue NagahashibookPart Tontura, Vertigem e Convulsão(2015) MARIE, Suely Kazue NagahashibookPart Cefaleias(2015) MARIE, Suely Kazue Nagahashi- Mass Spectrometry-Based Characterization of Protein Aggregates in Tissues and Biofluids(2024) MACEDO-DA-SILVA, J.; ROSA-FERNANDES, L.; SANTIAGO, V. F.; BLANES, C. A.; MARIE, S. K. N.; PALMISANO, G.Protein aggregation is a common mechanism in multiple neurodegenerative and heart diseases and the accumulation of proteins in aggregates is toxic to cells, causing injury and death. The degree of protein aggregation directly correlates with the severity of the disease. Misfolded proteins present thermodynamic barriers that culminate in the loss of structure and function and the exposure of hydrophobic residues. The exposure of hydrophobic residues is the driving force behind protein aggregation, as it reduces surface free energy and increases the propensity for the formation of large insoluble aggregates. Exploring the protein content of aggregates is fundamental to understanding their formation mechanism and pathophysiological effects. We demonstrate here a method for isolating aggregated protein content in human plasma and mouse brain samples. The samples were characterized by mass spectrometry analysis, transmission electron microscopy, and western blotting. We report the identification of proteins associated with neurodegenerative diseases in the isolated pellets. The western blotting analyses of the isolated pellet showed the positivity for CD89 and CD63, consolidated markers of exosomes, confirming the presence of exosomes within the pellet but not in the supernatant in human plasma. Notably, the concomitant isolation of exosomes together with the protein aggregates was feasible starting from 200 μL of human plasma. Moreover, the presented methodology separated albumin from the aggregated pellet, allowing identification of larger diversity of proteins through mass spectrometry analysis. © The Author(s), under exclusive license to Springer Nature Switzerland AG 2024.
- Isolation of Extracellular Vesicles Using Titanium Dioxide Microspheres(2024) SANTIAGO, V. F.; ROSA-FERNANDES, L.; MACEDO-DA-SILVA, J.; ANGELI, C. B.; MULE, S. N.; MARINHO, C. R. F.; TORRECILHAS, A. C.; MARIE, S. N. K.; PALMISANO, G.Extracellular vesicles (EVs) are bilayer membrane particles released from several cell types to the extracellular environment. EVs have a crucial role in cell-cell communication, involving different biological processes in health and diseases. Due to the potential of biomarkers for several diseases as diagnostic and therapeutic tools, it is relevant to understand the biology of the EVs and their content. One of the current challenges involving EVs is regarding the purification method, which is a critical step for EV’s functional and characterization studies. Ultracentrifugation is the most used method for EV isolation, where the nanoparticles are separated in sequential centrifugation to isolate the EVs based on their size. However, for viscous biofluids such as plasma, there is a co-isolation of the most abundant proteins, which can impair the EV’s protein identification due to the low abundance of these proteins and signal suppression by the most abundant plasma proteins. Emerging techniques have gained attention in recent years. Titanium dioxide (TiO2) is one of the most promising techniques due to its property for selective isolation based on the interaction with phospholipids in the EV membrane. Using a small amount of TiO2 beads and a low volume of plasma, it is possible to isolate EVs with reduced plasma protein co-isolation. This study describes a comprehensive workflow for the isolation and characterization of plasma extracellular vesicles (EVs) using mass spectrometry-based proteomics techniques. The aim of this chapter is describe the EV isolation using TiO2 beads enrichment and high-throughput mass spectrometry techniques to efficiently identify the protein composition of EVs in a fast and straightforward manner. © The Author(s), under exclusive license to Springer Nature Switzerland AG 2024.
- Exploring COVID-19 pathogenesis on command-line: A bioinformatics pipeline for handling and integrating omics data(2022) MACEDO-DA-SILVA, J.; COUTINHO, J. V. P.; ROSA-FERNANDES, L.; MARIE, S. K. N.; PALMISANO, G.Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was first identified in late 2019 in Wuhan, China, and has proven to be highly pathogenic, making it a global public health threat. The immediate need to understand the mechanisms and impact of the virus made omics techniques stand out, as they can offer a holistic and comprehensive view of thousands of molecules in a single experiment. Mastering bioinformatics tools to process, analyze, integrate, and interpret omics data is a powerful knowledge to enrich results. We present a robust and open access computational pipeline for extracting information from quantitative proteomics and transcriptomics public data. We present the entire pipeline from raw data to differentially expressed genes. We explore processes and pathways related to mapped transcripts and proteins. A pipeline is presented to integrate and compare proteomics and transcriptomics data using also packages available in the Bioconductor and providing the codes used. Cholesterol metabolism, immune system activity, ECM, and proteasomal degradation pathways increased in infected patients. Leukocyte activation profile was overrepresented in both proteomics and transcriptomics data. Finally, we found a panel of proteins and transcripts regulated in the same direction in the lung transcriptome and plasma proteome that distinguish healthy and infected individuals. This panel of markers was confirmed in another cohort of patients, thus validating the robustness and functionality of the tools presented. © 2022 Elsevier Inc.