The role of extracellular vesicles (EVs) proteome in diffuse large B-cell lymphoma (DLBCL) pathology, subclassification, and patient screening is unexplored. We analyzed by state-of-the-art mass spectrometry the whole cell and secreted extracellular vesicles (EVs) proteomes of different molecular subtypes of DLBCL, germinal center B cell (GCB subtype), and activated B cell (ABC subtype). After quality control assessment, we compared whole-cell and secreted EVs proteomes of the two cell-of-origin (COO) categories, GCB and ABC subtypes, resulting in 288/1115 significantly differential expressed proteins from the whole-cell proteome and 228/608 proteins from EVs (adjust p-value < 0.05/p-value < 0.05). In our preclinical model system, we demonstrated that the EV proteome and the whole-cell proteome possess the capacity to separate cell lines into ABC and GCB subtypes. KEGG functional analysis and GO enrichment analysis for cellular component, molecular function, and biological process of differential expressed proteins (DEP) between ABC and GCB EVs showed a significant enrichment of pathways involved in immune response function. Other enriched functional categories for DEPs constitute cellular signaling and intracellular trafficking such as B-cell receptor (BCR), Fc_gamma R-mediated phagocytosis, ErbB signaling, and endocytosis. Our results suggest EVs can be explored as a tool for patient diagnosis, follow-up, and disease monitoring. Finally, this study proposes novel drug targets based on highly expressed proteins, for which antitumor drugs are available suggesting potential combinatorial therapies for aggressive forms of DLBCL. Data are available via ProteomeXchange with identifier PXD028267.
Extracellular Vesicles/metabolism , Lymphoma, Large B-Cell, Diffuse/pathology , Proteome/analysis , Proteomics/methods , B-Lymphocytes/metabolism , Cell Line, Tumor , Germinal Center/cytology , Germinal Center/metabolism , Humans , Lymphoma, Large B-Cell, Diffuse/metabolism , Mass Spectrometry
Depression and anxiety have been reported as the major neuropsychiatric consequences following stroke. Minocycline, a neuroprotective drug has minimized depressive symptoms in patients with major depressive disorders and anxiety-like symptoms. In addition, minocycline demonstrated efficacy and seemed a promising neuroprotective agent in acute stroke patients. The present studied evaluated the effects of minocycline treatment on the depression and anxiety-like behaviors, brain damage and expression of inflammatory and neuroprotective mediators after transient global cerebral ischemia in C57BL/6 mice. Brain ischemia was induced by bilateral occlusion of the common carotids (BCCAo) for 25 min and subsequent reperfusion. Sham and BCCAo animals received minocycline at a dose of 30 mg/kg by intraperitoneal injection during 14 days. The locomotor activity, depression and anxiety-like behaviors were assessed by open field, forced swim and elevated plus maze tests, respectively. Then, the brains were removed and processed to evaluate brain damage by histological and morphometric analysis, hippocampal neurodegeneration using Fluoro-Jade C histochemistry, microglial activity using iba-1 immunohistochemistry, brain levels of TNF, IFN-γ, IL-6, IL-10, IL-12p70 and CCL2 by CBA, CX3CL1 and BDNF by ELISA assays. The animals developed depression and anxiety-like behaviors post-stroke and minocycline treatment prevented those neurobehavioral changes. Moreover, minocycline-treated BCCAo animals showed less intense brain damage in the cerebral cortex, brainstem and cerebellum as well as significantly reduced hippocampal neurodegeneration. BCCAo groups exhibited up-regulation of some cytokines at day 14 after ischemia and brain levels of CX3CL1 and BDNF remained unaltered. Our data indicate that the depression and anxiety-like behavioral improvements promoted by minocycline treatment might be related to its neuroprotective effect after brain ischemia in mice.
Anxiety/prevention & control , Depression/prevention & control , Ischemic Stroke/prevention & control , Minocycline/administration & dosage , Neuroprotective Agents/administration & dosage , Animals , Brain/drug effects , Brain/pathology , Encephalitis/prevention & control , Hippocampus/drug effects , Hippocampus/pathology , Ischemic Stroke/pathology , Male , Mice, Inbred C57BL , Neurons/pathology
Through lateral transfer, extra-cellular vesicles (EVs) transport their DNA, miRNA, mRNA and proteins such as enzymes mediating drug resistance, transporters as well as growth factors to neighboring cells. By virtue of this horizontal transfer, EVs potentially regulate cell growth, migration, angiogenesis and metastasis and increase tissue permeability in cancer. Furthermore, EVs regulate immune factors and allow the tumor cells to evade immune recognition and cell death. To explore if the proteomes of exosomes support functional transfer of cancer hallmarks, in this meta-analysis, we compared EVs and whole cell proteomes from the NCI-60 human tumor cell line panel. We observed a subgroup of proteins in each cancer hallmark signature as highly abundant and consistently expressed in EVs from all cell lines. Among these were oncoproteins frequently targeted in cancer therapies whose presence on EVs could potentially render therapies less effective by serving as decoys.
Exosomes/metabolism , Extracellular Vesicles/metabolism , Neoplasms/metabolism , Oncogene Proteins/metabolism , Proteome/metabolism , Proteomics/methods , Cell Line, Tumor , Humans , Neoplasms/pathology , Signal Transduction , Tumor Suppressor Proteins/metabolism
Stroke is one of the most frequent causes of death and disability worldwide leading to a significant clinical and socioeconomic burden. Although different mechanisms are involved in the pathogenesis of stroke, inflammatory response occurs after ischemia and contributes to the expansion of brain injury. Platelet-activating factor receptor (PAF) plays crucial roles in both physiological and pathological conditions in the brain. PAF receptor (PAFR) may be expressed on cellular and nuclear membranes of various cell types, especially leukocytes, platelets, endothelial cells, neuronal cells and microglia. Herein, using mice lacking the PAFR receptor (PAFR(-/-)), we investigate a potential role for this receptor during experimental transient global cerebral ischemia and reperfusion (BCCAo). In PAFR deficiency, we observed a significant improvement in the neurological deficits, which were associated with a reduction of brain infarcted area as evaluated by triphenyltetrazolium chloride (TTC). Moreover, a decrease in the percentage of necrotic cavities areas and in the frequency of ischemic neurons was also found by employing histometric analysis. In addition, in PAFR(-/-) mice there was prevention of caspase-3 activation and decreased vascular permeability and brain edema. Decreased brain levels of the cytokines tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta (IL-1beta) and the chemokine (C-X-C motif) ligand 1 (CXCL1) by ELISA were also detected in PAFR(-/-) BCCAo animals. Taken together, our results suggest that PAFR activation might be crucial for the global brain ischemia and reperfusion injury.
Ischemic Attack, Transient/metabolism , Platelet Membrane Glycoproteins/metabolism , Receptors, G-Protein-Coupled/metabolism , Reperfusion Injury/metabolism , Animals , Blood-Brain Barrier/physiopathology , Brain Infarction/etiology , Caspase 3/metabolism , Cytokines/metabolism , Disease Models, Animal , Gene Expression Regulation/genetics , Ischemic Attack, Transient/complications , Ischemic Attack, Transient/pathology , Male , Mice , Mice, Inbred C57BL , Mice, Knockout , Nervous System Diseases/etiology , Platelet Membrane Glycoproteins/genetics , Receptors, G-Protein-Coupled/genetics , Reperfusion Injury/pathology , Statistics, Nonparametric
Stroke is one of the most frequent causes of death and disability worldwide causing a major clinical and socioeconomic impact. Although the pathophysiology of brain ischemia and reperfusion is complex, the inflammatory process plays an important role in pathogenesis, contributing to the expansion of brain injury. The 5-lipoxygenase (5-LOX) is a key enzyme in the biosynthesis of the leukotrienes and has been implicated and in the central nervous system (CNS) disorders such as Alzheimer's disease and acute ischemic stroke. Zileuton, a selective 5-LOX inhibitor, has antiinflammatory properties and exerts an inhibitory effect on inflammatory diseases. The objective of this study was to evaluate the effects of blocking 5-LOX activity in a murine model of transient and global brain ischemia. Zileuton improved neurological deficits and significantly decrease volume and density of lesion, compared to vehicle-ischemic animals measured by magnetic resonance imaging (MRI). In addition, the blockage of 5-LOX reduced infarct area and histopathological changes. Furthermore, by enzyme immunoassay (ELISA) increased brain levels of tumor necrosis factor-alpha (TNFalpha), interferon-gamma (IFN-gamma), interleukin-1beta (IL-1beta), interleukin-6 (IL-6), chemokine (C-X-C motif) ligand 1 (CXCL1), chemokine (C-C motif) ligand 3 (CCL3) and chemokine (C-C motif) ligand 5 (CCL5) were detected in the vehicle-ischemic group, whereas in Zileuton-ischemic group presented reduction of these mediators. The concentration of the antiinflammatory cytokine interleukin-10 (IL-10) was increased after 5-LOX inhibition. Our results suggest that Zileuton decreases brain damage and reduces inflammatory cytokines expression in the CNS which contributes, at least in part, to improve the neurological outcome of brain ischemia.
Anti-Inflammatory Agents, Non-Steroidal/therapeutic use , Brain Infarction/drug therapy , Encephalitis/drug therapy , Hydroxyurea/analogs & derivatives , Nervous System Diseases/complications , Analysis of Variance , Animals , Brain Infarction/etiology , Carotid Stenosis/complications , Cytokines/genetics , Cytokines/metabolism , Disease Models, Animal , Down-Regulation/drug effects , Encephalitis/etiology , Enzyme-Linked Immunosorbent Assay , Hydroxyurea/therapeutic use , Magnetic Resonance Imaging , Male , Mice , Mice, Inbred C57BL , Nervous System Diseases/etiology
OBJETIVO: Avaliar clínica e radiograficamente o ângulo de carregamento do cotovelo (ACC) determinando uma curva de normalidade de acordo com faixas etárias (da infância à maturidade esquelética) e comparar as medidas clínicas e radiográficas. MÉTODOS: Avaliamos 510 indivíduos (1020 cotovelos), com idades entre 1 e 18 anos, distribuídos em grupos de 30 conforme faixas etárias, com intervalo de 1 ano. Excluímos os portadores de: fraturas do cotovelo, sequelas, malformações, doenças genéticas, afecções inflamatórias e frouxidão ligamentar. Mensuramos clinicamente o ACC bilateralmente com goniômetro, onde obtivemos duas medidas por dois examinadores onde média destas foi considerada. Realizamos radiografias ântero-posteriores dos cotovelos e aferimos os ângulos formados pelos eixos do úmero e da ulna. Todos os dados foram analisados estatisticamente pelo teste t student. RESULTADOS: Determinamos uma curva de normalidade onde observamos aumento deste parâmetro conforme a progressão da idade. Não observamos diferença significante entre as medidas clínicas e radiográficas. CONCLUSÃO: A média do ângulo de carregamento para o sexo feminino foi 12,78º ± 5,35 e para o masculino 11,20º ± 4,45. Este valor aumenta progressivamente da infância até os 16 anos quando observamos estabilização. Não houve diferença estatística significante das medidas clínicas e radiográficas.
OBJECTIVE: This paper has the purpose of evaluate the elbow carrying angle by clinic and radiographic examination in normal children and determine the range of normality according to age from childhood to skeletal maturity and also check if there is a statistically significant difference between the clinical and radiographic measurements. METHODS: We evaluated 510 persons with ages varying from 1 to 18 years distributed in groups with 30 subjects according to the age group with 1-year interval. We performed radiographic examination of the elbow and measured the angle formed by the long axis of the humerus and ulna. The data were statistically analyzed using the student t-test. RESULTS: We determined a normal curve of the study population where there was an increase of this parameter with the progression of age. No statistically significant difference between the clinical and radiographic measures. CONCLUSION: The average of the elbow carrying angle was 12,78 ± 5,35 degrees for females and 11,20 ± 4,45 degrees for males. This values increase progressively from childhood until 16 years when we notice stabilization. There was no statistical difference between the clinical and radiographic measurements.
Humans , Male , Female , Infant , Child, Preschool , Child , Adolescent , Arthroplasty, Replacement, Elbow , Anthropometry/methods , Elbow Joint , Age Distribution , Brazil , Cross-Sectional Studies , Joint Instability/pathology
Os auto-anticorpos antiantígenos citoplasmáticos de neutrófilos (ANCA) têm importante associação ao diagnóstico e possível monitoramento de uma significante parcela de doenças auto-imunes. A Proteinase 3 é o principal antígeno presente no tipo demarcação citoplasmática granular difusa (c-ANCA), e a Mieloperoxidase na marcação perinuclear (p-ANCA) em ensaios de imunofluorescência indireta (IFI) em neutrófilos fixados em etanol. A vasculite sistêmica compreende uma série de síndromes caracterizadas por dividir uma base histopatológica comum: inflamação nos vasos sangüíneos resultando em obstrução vascular com subseqüente isquemia e enfartamento tissular. A vasculite constitui um grupo heterogêneo de doenças que possuem como característica comum à inflamação destrutiva da parede de vasos sangüíneos. O papel potencializador de ANCA sobre as lesões é descrito em estágios iniciais de vasculite sistêmica pelo fato deste promover o recrutamento e adesões entre neutrófilos e células endoteliais.O presente trabalho teve como objetivo enfocar o papel do c-ANCA e p-ANCA no diagnóstico laboratorial de Vasculite Sistêmica e o levantamento de exames efetuados no período de 16/04/04 a 20/04/06 para estes mesmos marcadores no laboratório de ImunologiaClínica do Hospital Universitário Polydoro Ernani de São Thiago. Neste levantamento pudemos constatar uma reduzida parcela de resultados positivos para c-ANCA (15%) e p-ANCA (39%), fato este devido à larga gama de doenças com sintomatologias semelhantes às doenças relacionadas à ANCA, possuindo, porém ANCA negativo.A avaliação clínica é importante no manejo de doenças auto-imunes, mas o laboratório representa papel decisivo na avaliação dessas doenças. Testes laboratoriais auxiliam ao estabelecer o diagnóstico, na monitorização do curso da doença, na predição de sua evolução,na decisão acerca da terapêutica, na avaliação da resposta à terapia e também para o estudo da etiologia ou patogênese das doenças auto-imunes
Humans , Antibodies, Antineutrophil Cytoplasmic , Autoimmune Diseases , Clinical Laboratory Techniques , Vasculitis
