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1.
Blood Res ; 58(1): 20-27, 2023 Mar 31.
Artigo em Inglês | MEDLINE | ID: mdl-36632684

RESUMO

Background: Leukemia is a neoplasm with high incidence and mortality rates. Mitotic death has been observed in tumor cells treated with chemotherapeutic agents. Ras family proteins participate in the transduction of signals involved in different processes, such as proliferation, differentiation, survival, and paradoxically, initiation of cell death. Methods: This study investigated the effect of H-Ras expression on human T-cell acute lymphoblastic leukemia MOLT-4 cells. Cells were electroporated with either wild-type (Raswt) or oncogenic mutant in codon 12 exon 1 (Rasmut) versions of H-Ras gene and stained for morphological analysis. Cell viability was assessed using trypan blue staining and cell cycle analysis using flow cytometry. H-Ras gene expression was determined using quantitative real-time reverse transcription polymerase chain reaction. The t, ANOVA, and Scheffe tests were used for statistical analysis. Results: Human T-cell acute lymphoblastic leukemia MOLT-4 cells showed nuclear fragmentation and presence of multiple nuclei and micronuclei after transfection with either wt or mutant H-Ras genes. Cell cycle analysis revealed a statistically significant increase in cells in the S phase when transfected with either wt (83.67%, P<0.0005) or mutated (81.79%, P<0.0001) H-Ras genes. Although similar effects for both versions of H-Ras were found, cells transfected with the mutated version died at 120 h of mitotic catastrophe. Conclusion: Transfection of human T-cell acute lymphoblastic leukemia MOLT-4 cells with either normal or mutated H-Ras genes induced alterations in morphology, arrest in the S phase, and death by mitotic catastrophe.

2.
Pathogens ; 13(1)2023 Dec 30.
Artigo em Inglês | MEDLINE | ID: mdl-38251344

RESUMO

Tuberculosis (TB) of the central nervous system (CNS) is a lethal and incapacitating disease. Several studies have been performed to understand the mechanism of bacterial arrival to CNS, however, it remains unclear. Although the interaction of the host, the pathogen, and the environment trigger the course of the disease, in TB the characteristics of these factors seem to be more relevant in the genesis of the clinical features of each patient. We previously tested three mycobacterial clinical isolates with distinctive genotypes obtained from the cerebrospinal fluid of patients with meningeal TB and showed that these strains disseminated extensively to the brain after intratracheal inoculation and pulmonary infection in BALB/c mice. In this present study, BALB/c mice were infected through the intranasal route. One of these strains reaches the olfactory bulb at the early stage of the infection and infects the brain before the lungs, but the histological study of the nasal mucosa did not show any alteration. This observation suggests that some mycobacteria strains can arrive directly at the brain, apparently toward the olfactory nerve after infecting the nasal mucosa, and guides us to study in more detail during mycobacteria infection the nasal mucosa, the associated connective tissue, and nervous structures of the cribriform plate, which connect the nasal cavity with the olfactory bulb.

3.
Annu Int Conf IEEE Eng Med Biol Soc ; 2021: 4072-4078, 2021 11.
Artigo em Inglês | MEDLINE | ID: mdl-34892124

RESUMO

In this work, we develop a patch-level training approach and a task-driven intensity-based augmentation method for deep-learning-based segmentation of motion-corrected perfusion cardiac magnetic resonance imaging (MRI) datasets. Further, the proposed method generates an image-based uncertainty map thanks to a novel spatial sliding-window approach used during patch-level training, hence allowing for uncertainty quantification. Using the quantified uncertainty, we detect the out-of-distribution test data instances so that the end-user can be alerted that the test data is not suitable for the trained network. This feature has the potential to enable a more reliable integration of the proposed deep learning-based framework into clinical practice. We test our approach on external MRI data acquired using a different acquisition protocol to demonstrate the robustness of our performance to variations in pulse-sequence parameters. The presented results further demonstrate that our deep-learning image segmentation approach trained with the proposed data-augmentation technique incorporating spatiotemporal (2D+time) patches is superior to the state-of-the-art 2D approach in terms of generalization performance.


Assuntos
Aprendizado Profundo , Coração/diagnóstico por imagem , Imageamento por Ressonância Magnética , Perfusão , Incerteza
4.
J Circadian Rhythms ; 19: 9, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34326881

RESUMO

The purpose of this study was to analyze the light-dark variations in the concentrations of several neurotransmitters in the lumbar spinal cord of rats. Six groups of male Wistar rats were exposed to a 12 h light-12 h dark cycle for 70 days. At different time points of the experimental day (8, 12, 16, 20, 24 and 4 h), one of the groups of rats was randomly selected to be sacrificed, and the spinal cords were removed. The gamma-aminobutyric acid (GABA), glutamate (GLU), dopamine, serotonin, epinephrine (E), and norepinephrine (NE) levels in each extracted spinal cord were measured with high-pressure liquid chromatography (HPLC)-EQ and HPLC-fluorescence systems. Our results indicate that the spinal concentrations of GABA and GLU showed sinusoidal variation in a 24 h cycle, with the highest peak in the dark period (~20 h). Dopamine and serotonin also fluctuated in concentration but peaked in the light period (between 8 and 12 h), while E and NE concentrations showed no significant fluctuations. The possible relationship between neurotransmitter spinal concentration and sensitivity to pain and locomotor activity is discussed. It was concluded that most of the neurotransmitter levels in the lumbar spinal cord showed circadian fluctuations coupled to a light-dark cycle.

5.
J Immunol Res ; 2021: 5517856, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34007850

RESUMO

The mechanisms underlying the immunopathology of tuberculous meningitis (TBM), the most severe clinical form of extrapulmonary tuberculosis (TB), are not understood. It is currently believed that the spread of Mycobacterium tuberculosis (Mtb) from the lung is an early event that occurs before the establishment of adaptive immunity. Hence, several innate immune mechanisms may participate in the containment of Mtb infection and prevent extrapulmonary disease manifestations. Natural killer (NK) cells participate in defensive processes that distinguish latent TB infection (LTBI) from active pulmonary TB (PTB). However, their role in TBM is unknown. Here, we performed a cross-sectional analysis of circulating NK cellCID="C008" value="s" phenotype in a prospective cohort of TBM patients (n = 10) using flow cytometry. Also, we addressed the responses of memory-like NK cell subpopulations to the contact with Mtb antigens in vitro. Finally, we determined plasma levels of soluble NKG2D receptor ligands in our cohort of TBM patients by enzyme-linked immunosorbent assay (ELISA). Our comparative groups consisted of individuals with LTBI (n = 11) and PTB (n = 27) patients. We found that NK cells from TBM patients showed lower absolute frequencies, higher CD69 expression, and poor expansion of the CD45RO+ memory-like subpopulation upon Mtb exposure in vitro compared to LTBI individuals. In addition, a reduction in the frequency of CD56brightCD16- NK cells characterized TBM patients but not LTBI or PTB subjects. Our study expands on earlier reports about the role of NK cells in TBM showing a reduced frequency of cytokine-producing cells compared to LTBI and PTB.


Assuntos
Células Matadoras Naturais/imunologia , Tuberculose Latente/imunologia , Mycobacterium tuberculosis/imunologia , Tuberculose Meníngea/imunologia , Tuberculose Pulmonar/imunologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Estudos Transversais , Citocinas/metabolismo , Feminino , Humanos , Imunidade Inata , Imunofenotipagem , Células Matadoras Naturais/metabolismo , Tuberculose Latente/sangue , Tuberculose Latente/microbiologia , Masculino , México , Pessoa de Meia-Idade , Estudos Prospectivos , Tuberculose Meníngea/sangue , Tuberculose Meníngea/microbiologia , Tuberculose Pulmonar/sangue , Tuberculose Pulmonar/microbiologia , Adulto Jovem
6.
J Matern Fetal Neonatal Med ; 34(14): 2323-2333, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31537145

RESUMO

PURPOSE: Neonatal sepsis is an important public health concern worldwide due to its immediate lethality and long-term morbidity rates, Clinical evaluation and laboratory analyses are indispensable for diagnosis of neonatal sepsis. However, assessing multiple biomarkers in neonates is difficult due to limited blood availability. The aim is to investigate if the neonatal sepsis in preterm could be identified by multiparameter analysis with flow cytometry. MATERIALS AND METHODS: The expression of activation-related molecules was evaluated by flow cytometry in newborn with or without risk factors for sepsis. RESULTS: Our analysis revealed that several markers could be useful for sepsis diagnosis, such as CD45RA, CD45RO, or CD71 on T cells; HLA-DR on NKT or classic monocytes, and TREM-1 on non-classic monocytes or neutrophils. However, ROC analysis shows that the expression of CD45RO on T lymphocytes is the only useful biomarker for diagnosis of neonatal late-onset sepsis. Also, decision tree analyses showed that CD45RO plus CD27 could help differentiate the preterm septic neonates from those with risk factors. CONCLUSIONS: Our study shows a complementary and practical strategy for biomarker assessment in neonatal sepsis.


Assuntos
Sepse Neonatal , Sepse , Biomarcadores , Citometria de Fluxo , Humanos , Recém-Nascido , Monócitos , Sepse Neonatal/diagnóstico , Sepse/diagnóstico
7.
J Med Virol ; 93(4): 2029-2038, 2021 04.
Artigo em Inglês | MEDLINE | ID: mdl-32986250

RESUMO

SARS-CoV-2 infection is causing a pandemic disease that is reflected in challenging public health problems worldwide. Human leukocyte antigen (HLA)-based epitope prediction and its association with disease outcomes provide an important base for treatment design. A bioinformatic prediction of T cell epitopes and their restricted HLA Class I and II alleles was performed to obtain immunogenic epitopes and HLA alleles from the spike protein of the severe acute respiratory syndrome coronavirus 2 virus. Also, a correlation with the predicted fatality rate of hospitalized patients in 28 states of Mexico was done. Here, we describe a set of 10 highly immunogenic epitopes, together with different HLA alleles that can efficiently present these epitopes to T cells. Most of these epitopes are located within the S1 subunit of the spike protein, suggesting that this area is highly immunogenic. A statistical negative correlation was found between the frequency of HLA-DRB1*01 and the fatality rate in hospitalized patients in Mexico.


Assuntos
Apresentação de Antígeno , COVID-19 , Cadeias HLA-DRB1/imunologia , Glicoproteína da Espícula de Coronavírus/imunologia , COVID-19/imunologia , COVID-19/mortalidade , Biologia Computacional , Epitopos de Linfócito T/imunologia , Variação Genética , Hospitalização , Humanos , México , Estrutura Terciária de Proteína , SARS-CoV-2/imunologia
8.
Front Immunol ; 11: 582414, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33117393

RESUMO

Natural killer (NK) cells participate in immunity against several pathogens by exerting cytotoxic and cytokine-production activities. Some NK cell subsets also mediate recall responses that resemble memory of adaptive lymphocytes against antigenic and non-antigenic stimuli. The C-X-C motif chemokine receptor 6 (CXCR6) is crucial for the development and maintenance of memory-like responses in murine NK cells. In humans, several subsets of tissue-resident and circulating NK cells with different functional properties express CXCR6. However, the role of CXCR6+ NK cells in immunity against relevant human pathogens is unknown. Here, we addressed whether murine and human CXCR6+ NK cells respond to antigens of Mycobacterium tuberculosis (Mtb). For this purpose, we evaluated the immunophenotype of hepatic and splenic CXCR6+ NK cells in mice exposed to a cell-wall (CW) extract of Mtb strain H37Rv. Also, we characterized the expression of CXCR6 in peripheral NK cells from active pulmonary tuberculosis (ATB) patients, individuals with latent TB infection (LTBI), and healthy volunteer donors (HD). Furthermore, we evaluated the responses of CXCR6+ NK cells from HD, LTBI, and ATB subjects to the in vitro exposure to CW preparations of Mtb H37Rv and Mtb HN878. Our results showed that murine hepatic CXCR6+ NK cells expand in vivo after consecutive administrations of Mtb H37Rv CW to mice. Remarkably, pooled hepatic and splenic, but not isolated splenic NK cells from treated mice, enhance their cytokine production capacity after an in vitro re-challenge with H37Rv CW. In humans, CXCR6+ NK cells were barely detected in the peripheral blood, although slightly significative increments in the percentage of CXCR6+, CXCR6+CD49a-, CXCR6+CD49a+, and CXCR6+CD69+ NK cells were observed in ATB patients as compared to HD and LTBI individuals. In contrast, the expansion of CXCR6+CD49a- and CXCR6+CD69+ NK cells in response to the in vitro stimulation with Mtb H37Rv was higher in LTBI individuals than in ATB patients. Finally, we found that Mtb HN878 CW generates IFN-γ-producing CXCR6+CD49a+ NK cells. Our results demonstrate that antigens of both laboratory-adapted and clinical Mtb strains are stimulating factors for murine and human CXCR6+ NK cells. Future studies evaluating the role of CXCR6+ NK cells during TB are warranted.


Assuntos
Antígenos de Bactérias/imunologia , Células Matadoras Naturais/imunologia , Tuberculose Latente/imunologia , Mycobacterium tuberculosis/fisiologia , Tuberculose Pulmonar/imunologia , Animais , Células Cultivadas , Feminino , Humanos , Imunofenotipagem , Interferon gama/metabolismo , Ativação Linfocitária , Camundongos , Camundongos Endogâmicos C57BL , Receptores CXCR6/metabolismo
9.
Rheumatology (Oxford) ; 59(4): 879-888, 2020 04 01.
Artigo em Inglês | MEDLINE | ID: mdl-31578573

RESUMO

OBJECTIVES: Expression of α4ß7 integrin can identify gut-homing immune cells. This study aimed to determine the expression of Toll-like receptor 2 (TLR2) and TLR4 in α4ß7-positive leukocytes of patients with axial SpA (axSpA). METHODS: We analysed the frequencies of α4ß7-positive T cells, Tγδ cells and monocytes in 14 patients with axSpA and 14 healthy controls, together with the expression of TLR2 and TLR4 by flow cytometry. Also, the concentration of faecal calprotectin was measured in all patients and controls. RESULTS: We found significantly higher percentages of α4ß7-positive T (P = 0.026) and Tγδ cells (P = 0.0118) in the patients with axSpA than in controls; these cells showed differential expression of TLR2 and TLR4 when compared with α4ß7-negative cells. Such differences were not correlated with disease activity or faecal calprotectin concentration. CONCLUSION: There is an increase in circulating α4ß7-positive T and Tγδ cells in patients with axSpA. These cells differentially express TLR2 and TLR4.


Assuntos
Monócitos/metabolismo , Espondiloartropatias/metabolismo , Subpopulações de Linfócitos T/metabolismo , Linfócitos T/metabolismo , Receptor 2 Toll-Like/metabolismo , Receptor 4 Toll-Like/metabolismo , Adulto , Estudos de Casos e Controles , Feminino , Humanos , Integrinas/metabolismo , Complexo Antígeno L1 Leucocitário/metabolismo , Masculino , Receptores de Antígenos de Linfócitos T gama-delta/metabolismo
10.
Sci Rep ; 9(1): 2152, 2019 02 15.
Artigo em Inglês | MEDLINE | ID: mdl-30770858

RESUMO

Atmospheric rivers (ARs) cause heavy precipitation and flooding in the coastal areas of many mid-latitude continents, and thus the atmospheric processes associated with the AR have been intensively studied in recent years. However, AR-associated ocean variability and air-sea fluxes have received little attention because of the lack of high-resolution ocean data until recently. Here we demonstrate that typical ARs can generate strong upper ocean response and substantial air-sea fluxes using a high-resolution (1/12°) ocean reanalysis. AR events observed during the CalWater 2015 field campaign generate large-scale on-shore currents that hit the coast, generating strong narrow northward jets along the west coast of North America, in association with a substantial rise of sea level at the coast. In the open ocean, the AR generates prominent changes of mixed layer depth, especially south of 30°N due to the strong surface winds and air-sea heat fluxes. The prominent cooling of SST is observed only in the vicinity of AR upstream areas primarily due to the large latent heat flux. Using a long-term AR dataset, composite structure and variations of upper ocean and air-sea fluxes are presented, which are consistent with those found in the events during CalWater 2015.

11.
Int J Oncol ; 52(4): 1246-1254, 2018 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-29436616

RESUMO

Pentoxifylline (PTX), a xanthine family molecule and simvastatin (SIM), an anti-hypercholesterolemic agent, have recently been considered as sensitizers to chemotherapy and radiotherapy. The present in vitro study evaluated their antitumor synergistic effects on MDA­MB­231 breast cancer cells characterized by the triple­negative phenotype (TNP). The anti-proliferative effects of these two agents were evaluated by MTT and clonogenic assays. Cell cycle progression was examined using propidium iodide staining. Apoptosis was investigated by Annexin V labeling, and by examining caspase 3 activity and DNA fragmentation. Autophagic vesicles and reactive oxygen species (ROS) levels were monitored by flow cytometry. Western blot analysis was performed to evaluate molecular targets. Our results revealed that when used alone, PTX and SIM exerted antitumor effects. Nevertheless, used in combination, the inhibition of cell proliferation was synergistically superior (80% vs 42%) than that observed following treatment with each agent alone after 48 h. PTX alone (0.5 mM) induced both apoptosis (25%) and autophagy (25%); however, when used in combination with SIM (0.5 µM), the balance between these processes was disrupted and the cells underwent apoptosis (>65%) as opposed to autophagy (<13%). This imbalance was associated with an increase in ERK1/2 and AKT activation, but not with an increase in mTOR phosphorylation, and with the suppression of the NF-κB pathway. In addition, in the cells treated with both agents, almost 78% of the cells were arrested at the G0/G1 phase and lost their colony-forming ability (38±5%) compared to the cells treated with PTX alone (115±5%). On the whole, these results suggest that the induction of autophagy may be a protective mechanism preventing MDA­MB­231 cancer cell death. The combined use of PTX and SIM may drive dormant autophagic cancer cells to undergo apoptosis and thus this may be a novel treatment strategy for breast cancer characterized by the TNP.


Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/farmacologia , Apoptose/efeitos dos fármacos , Pentoxifilina/farmacologia , Sinvastatina/farmacologia , Neoplasias de Mama Triplo Negativas/patologia , Autofagia/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sinergismo Farmacológico , Feminino , Humanos
12.
Biol Res ; 50(1): 42, 2017 Dec 21.
Artigo em Inglês | MEDLINE | ID: mdl-29268779

RESUMO

BACKGROUND: Fresh frozen plasma (FFP) administration may increase the risk of nosocomial infections in parallel with the development of immune modulation. This could be driven by soluble mediators, possibly influencing the in vitro activation of human U937 monocyte cells, in a manner dependent on the age of the donors. METHODS: FFP donors were stratified into groups of 19-30 years, 31-40 years or 41-50 years, and U937 cells were cultured with FFP (alone or plus lipopolysaccharide-LPS) for 24 h. Both in FFP and supernatants, TNF, IL-1ß, IL-6, and IL-10 levels were measured by ELISA. Additionally, CD11B, TLR2, and CASP3 gene expression were measured by qtPCR in U937 cells. Total phagocytic activity was also assayed. RESULTS: Elevated IL-10, but low TNF and IL-1ß levels were measured in FFP from individuals aged 19-40 years, whereas in individuals aged 41-50 years FFP were characterized by equalized TNF and IL-10 levels. Elevated IL-6 levels were found in all FFP samples, especially in those from the oldest individuals. FFP stimulation was associated with striking modifications in cytokine production in an age-dependent way. Exposure to FFP attenuates the response to LPS. TLR2 and CD11B expression were enhanced regardless of the age of plasma donors, although CASP3 expression was increased only when FFP from individuals aged 19-40 years were tested. Phagocytosis decreased after exposure to FFP regardless of donor age. CONCLUSION: Our results suggest that soluble mediators in FFP may modulate the functioning of monocytes. Interestingly, this effect appears to be partially influenced by the age of donors.


Assuntos
Doadores de Sangue , Citocinas/imunologia , Monócitos/imunologia , Plasma/imunologia , Células U937/imunologia , Adulto , Fatores Etários , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Interleucina-10/metabolismo , Interleucina-1beta/metabolismo , Interleucina-6/metabolismo , Masculino , Pessoa de Meia-Idade , Monócitos/fisiologia , Fator de Necrose Tumoral alfa/metabolismo , Adulto Jovem
13.
Front Immunol ; 8: 967, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28855906

RESUMO

Natural killer (NK) cells are lymphocytes of the innate immune system, which play an important role in the initial defense against a wide variety of pathogens, including viruses and intracellular bacteria. NK cells produce cytokines that enhance immune responses directed toward pathogens and also exert cytotoxic activity against infected cells, thereby eliminating the reservoir of infection. Their role in defense against Mycobacterium tuberculosis (Mtb) has been recently studied, and there is increasing evidence that highlight the importance of NK cell function during pulmonary tuberculosis (PTB), especially in the absence of optimal T-cell responses. Additionally, in the last years, it has been observed that NK cells mediate secondary responses against antigens to which they were previously exposed, an ability classically attributed to lymphocytes of the adaptive branch of immunity. This phenomenon, called "innate memory," could have important implications in the efforts to develop therapies and vaccines to improve the initial phases of immune reactions against different microorganisms, especially those to which there is not yet available vaccines to prevent infection, as is the case for tuberculosis. Therefore, the possibility of inducing memory-like NK cells ready to act prior to contact with Mtb or during the earliest stages of infection becomes quite interesting. However, our understanding of the mechanisms of innate memory remains incomplete. Here, we review recent literature about the mechanisms involved in the formation and maintenance of NK cell memory and the role of these cells in the immune response during tuberculosis. Finally, we discuss if the current evidence is sufficient to substantiate that NK cells exert more rapid and robust secondary responses after consecutive encounters with Mtb.

14.
Int Angiol ; 36(3): 203-215, 2017 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-27386953

RESUMO

INTRODUCTION: Lower extremity peripheral artery disease (PAD) is increasing in prevalence in low- and middle-income countries creating a large health care burden. Clinical management may require substantial resources but little consideration has been given to which treatments are appropriate for less advantaged countries. EVIDENCE ACQUISITION: The aim of this review was to systematically appraise published data on the costs and effectiveness of PAD treatments used commonly in high-income countries, and for an international consensus panel to review that information and propose a hierarchy of treatments relevant to low- and middle-income countries. EVIDENCE SYNTHESIS: Pharmacotherapy for intermittent claudication was found to be expensive and improve walking distance by a modest amount. Exercise and endovascular therapies were more effective and exercise the most cost-effective. For critical limb ischemia, bypass surgery and endovascular therapy, which are both resource intensive, resulted in similar rates of amputation-free survival. Substantial reductions in cardiovascular events occurred with use of low cost drugs (statins, ACE inhibitors, anti-platelets) and smoking cessation. CONCLUSIONS: The panel concluded that, in low- and middle-income countries, cardiovascular prevention is a top priority, whereas a lower priority should be given to pharmacotherapy for leg symptoms and revascularisation, except in countries with established vascular units.


Assuntos
Extremidade Inferior/irrigação sanguínea , Doença Arterial Periférica/prevenção & controle , Doença Arterial Periférica/terapia , Amputação Cirúrgica , Análise Custo-Benefício , Tratamento Farmacológico , Procedimentos Endovasculares , Exercício Físico , Humanos , Pobreza/economia , Ensaios Clínicos Controlados Aleatórios como Assunto , Fatores de Risco
15.
Biol. Res ; 50: 42, 2017. tab, graf
Artigo em Inglês | LILACS | ID: biblio-950888

RESUMO

BACKGROUND: Fresh frozen plasma (FFP) administration may increase the risk of nosocomial infections in parallel with the development of immune modulation. This could be driven by soluble mediators, possibly influencing the in vitro activation of human U937 monocyte cells, in a manner dependent on the age of the donors. METHODS: FFP donors were stratified into groups of 19-30 years, 31-40 years or 41-50 years, and U937 cells were cultured with FFP (alone or plus lipopolysaccharide-LPS) for 24 h. Both in FFP and supernatants, TNF, IL-1ß, IL-6, and IL-10 levels were measured by ELISA. Additionally, CD11B, TLR2, and CASP3 gene expression were measured by qtPCR in U937 cells. Total phagocytic activity was also assayed. RESULTS: Elevated IL-10, but low TNF and IL-1ß levels were measured in FFP from individuals aged 19-40 years, whereas in individuals aged 41-50 years FFP were characterized by equalized TNF and IL-10 levels. Elevated IL-6 levels were found in all FFP samples, especially in those from the oldest individuals. FFP stimulation was associated with striking modifications in cytokine production in an age-dependent way. Exposure to FFP attenuates the response to LPS. TLR2 and CD11B expression were enhanced regardless of the age of plasma donors, although CASP3 expression was increased only when FFP from individuals aged 19-40 years were tested. Phagocytosis decreased after exposure to FFP regardless of donor age. CONCLUSION: Our results suggest that soluble mediators in FFP may modulate the functioning of monocytes. Interestingly, this effect appears to be partially influenced by the age of donors.


Assuntos
Humanos , Masculino , Feminino , Adulto , Pessoa de Meia-Idade , Adulto Jovem , Plasma/imunologia , Doadores de Sangue , Monócitos/imunologia , Citocinas/imunologia , Células U937/imunologia , Ensaio de Imunoadsorção Enzimática , Monócitos/fisiologia , Fatores Etários , Interleucina-6/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , Interleucina-10/metabolismo , Interleucina-1beta/metabolismo
16.
Mediators Inflamm ; 2015: 313140, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26504358

RESUMO

BALB/c strain mice are unique in that they develop murine plasmacytoma (MPC) as a consequence of the inflammation induced by pristane oil injection in the peritoneal cavity. In this work the Treg, Th17, B1, B2, and NHC lymphocyte populations from the peritoneal environment of BALB/c, the susceptible strain, and C57BL/6 mice, which do not develop MPC after oil treatment, were studied. Both oil-treated strains showed decreased levels of Th17 lymphocytes, no significant variation in Treg lymphocytes, and a drastic decrease of all B lymphocyte populations. However, only oil-induced BALB/c showed increased levels of natural helper cells (NHC) which could be important in the myeloma induction.


Assuntos
Linfócitos/citologia , Cavidade Peritoneal , Plasmocitoma/metabolismo , Linfócitos T Auxiliares-Indutores/citologia , Animais , Citocinas/metabolismo , Modelos Animais de Doenças , Feminino , Inflamação , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Óleos , Linfócitos T Reguladores/citologia , Terpenos/química , Células Th17/metabolismo
17.
Immunol Invest ; 42(4): 369-81, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23883202

RESUMO

The suppressor effect of T regulatory lymphocytes in co-cultures with T effector cells obtained by magnetic columns from healthy donors and activated by CD3/CD28 was measured by a proliferation assay using BrdU incorporation and an ELISA test. Tritiated thymidine incorporation was used as a reference since it is the gold standard for proliferation assays. Both methods were used simultaneously in the same samples in order to compare them. Correlation between them was statistically significant (p < 0.001). The purification using magnetic columns was very efficient since CD4⁺CD25⁺ cells were also FOXP3⁺ therefore; they were identified as suppressor T cells. The use of BrdU incorporation in suppression assays is an excellent method that avoids the use of radioactive contaminating materials.


Assuntos
Bioensaio , Bromodesoxiuridina/farmacologia , Subpopulações de Linfócitos T/imunologia , Linfócitos T Reguladores/imunologia , Adulto , Proliferação de Células , Células Cultivadas , Humanos , Subunidade alfa de Receptor de Interleucina-2/imunologia , Timidina/farmacologia , Trítio
18.
Allergy Asthma Proc ; 34(1): 84-92, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23406941

RESUMO

Asthma has been defined as a disease of chronic airway inflammation in which many cells and cellular products participate with variable degrees of airflow obstruction and hyperresponsiveness that lead to recurrent episodes of wheezing, breathlessness, chest tightness, and coughing. Prominent among these cellular elements are two cell types referred to as the invariant natural killer T (iNKT) cells and a subpopulation of T cells expressing the molecule CD161, which are both thought to play a role in the pathogenesis of asthma. Although the presence of iNKT and other CD161(+) cells in murine models has been associated with asthma, relatively few studies have been performed in the adult patient with asthma that have been often conflicting and even fewer studies are available in children. The present study was performed to investigate the peripheral blood frequencies of iNKT and CD161(+) T cells in children with asthma. A total of 35 children, 19 stable asthmatic patients, 6 who had experienced an asthmatic attack within 24 hours and had not received any treatment, and 10 healthy controls, aged 6-12 years, were enrolled in the study. iNKT and CD161(+) T-cell frequencies in blood were measured together with quantitative levels of IL-4 and interferon (IFN) γ using a cytofluorimetric approach. The results show that iNKT cells are increased in pediatric asthmatic patients undergoing exacerbations of asthma. These cells also produced less IFN-γ and more IL-4 than children with stable asthma and in healthy control children. These results suggest that iNKT cells might participate in the development of the asthmatic exacerbations. The increased production of IL-4 in conjunction with the decrease of IFN-γ may be mechanistically responsible, at least partially, for the heightening of the immunologic response leading to the asthmatic attack in children. Knowledge of these interactive mechanisms involving the iNKT cell and our understanding of its role in the exacerbation of asthma hold great promise in the development of better diagnostic predictive markers of disease progression as well as new forms of therapeutic interventions.


Assuntos
Asma/imunologia , Interferon gama/metabolismo , Interleucina-4/metabolismo , Células T Matadoras Naturais/imunologia , Antígenos CD4/metabolismo , Separação Celular , Criança , Progressão da Doença , Feminino , Citometria de Fluxo , Humanos , Ativação Linfocitária , Contagem de Linfócitos , Masculino , Subfamília B de Receptores Semelhantes a Lectina de Células NK/metabolismo
20.
Rev. argent. cir. cardiovasc. (Impresa) ; 9(3): 175-182, sept.-dic. 2011. ilus, tab
Artigo em Espanhol | LILACS | ID: lil-703265

RESUMO

La ateroembolia es una entidad poco conocida y cuyo pronóstico es sombrío, especialmente en aquellos cuadros de ateroembolia diseminada, pero también en casos de ateroembolia distal que provoca un síndrome inflamatorio de respuesta sistémica. Durante 14 años recopilamos 31 casos de ateroembolia, 5 de ellas postoperatorias y 26 casos con orígenes distintos: 20 debido a aorta Shaggy y 6 a otras causas. De las 20 con aorta Shaggy, 10 presentaban ateroembolia. 22 de los 26 casos no iatrogénicos fueron tratados por vía endovascular, con una mortalidad del 9%. De acuerdo a nuestras observaciones, comentamos nuestra conducta y las variaciones en el tratamiento, proponiendo una clasificación que contemple los casos asintomáticos para el tratamiento precoz de estas lesiones aórticas, antes de producir complicaciones severas por ateroembolias o síndromes isquémicos de los miembros inferiores. Asimismo, comentamos aquellos trucos que aprendimos con la observación de los distintos casos reseñados.


A ateroembolia é uma entidade pouco conhecida e cujo prognóstico é sombrio, especialmente em quadros de ateroembolia disseminada, mas que também ocorre em casos de ateroembolia distal, o que provoca uma síndrome inflamatória de resposta sistêmica. Durante 14 anos recolhemos 31 casos de ateroembolia. 5 deles, postoperatórias e 26 casos com origens diferentes: 20 devido à aorta Shaggy e 6 por outras causas. Das 20 com aorta Shaggy, 10 apresentavam ateroembolia. 22 dos 26 casos não iatrogênicos foram tratados via endovascular, com uma mortalidade de 9%. De acordo com as nossas observações, comentamos as condutas e variações no tratamento, propondo uma classificação que inclua os casos assintomáticos para o tratamento precoce destas lesões aórticas, antes de produzir complicações severas por ateroembolias ou síndromes isquêmicos dos miembros inferiores. Comentamos os truques que aprendemos com a observação dos diferentes casos selecionados.


Ateroembolism is not a very well known entity and its prognosis is poor, especially in cases of disseminated ateroembolism, and also in cases of distal ateroembolism which causes aninflammatory syndrome of systemic response. 31 cases of ateroembolism were registered during 14 years, 5 were post-surgical and 26 due to different etiologies: 20 due to shaggy aorta and 6 due to other reasons. Of the 20 cases with shaggy aorta, 10 presented ateroembolism. 22 of the 26 non-iatrogenic cases were treated with the endovascular approach with a 9% mortality. According to our observations, we discussed our approach and treatment options, suggesting a classification which includes asymptomatic cases for early treatment of these aortic lesions, before they induce severe ateroembolic complications or lower limb ischemic syndromes. Likewise, we discuss some tips learnt during the observation of the different cases commented.


Assuntos
Feminino , Aorta/cirurgia , Artéria Ilíaca , Embolia de Colesterol/cirurgia , Endarterectomia , Doenças Vasculares Periféricas/terapia , Fatores de Risco , Perna (Membro)/irrigação sanguínea
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