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Braz. J. Pharm. Sci. (Online) ; 58: e20096, 2022. tab, graf
Article in English | LILACS | ID: biblio-1403677


Abstract Dexchlorpheniramine is a first-generation classical antihistamine, clinically used to treat allergies. The main objective of our study was to evaluate the effects of the dexchlorpheniramine reference standard (DCPA Ref. St) and a pharmaceutical formula on DNA in human peripheral blood mononuclear cells (PBMCs). We exposed PBMCs to five different concentrations (0.5, 2.5, 5, 10, and 50 ng/mL) of DCPA Ref. St DCPA Ref. St and pharmaceutical formula in order to evaluate their cytotoxic, genotoxic, and mutagenic potential. The results showed that both dexchlorpheniramine formulations did not affect PBMC viability and CD3+, CD4+, or CD8+ lymphocyte subpopulations. The DCPA Ref. St and pharmaceutical formula neither induced genotoxic or mutagenic effects nor numerical or structural chromosomal alterations in PBMCs after 24 hours of exposure.

Humans , Leukocytes, Mononuclear/metabolism , Cytotoxicity, Immunologic , Drug Compounding , Genotoxicity , Mutagenicity Tests , DNA/analysis , Histamine Antagonists/adverse effects , Hypersensitivity/complications
Article in Chinese | WPRIM | ID: wpr-936325


OBJECTIVE@#To investigate the effect of triptolide (TPL) on inflammatory response and migration of fibroblast like synovial cells (FLS) in rheumatoid arthritis (RA-FLS) and the mechanism of circular noncoding RNA (circRNA) 0003353 for mediating this effect.@*METHODS@#We collected peripheral blood mononuclear cells (PBMCs) and serum samples from 50 hospitalized RA patients and 30 healthy individuals for detecting the expression of circRNA 0003353, immune and inflammatory indexes (ESR, CRP, RF, anti-CCP, IgA, IgG, IgM, C3, and C4) and DAS28 score. Cultured RA-FLS was treated with 10 ng/mL TPL and transfected with a circRNA 0003353 overexpression plasmid, and cell counting kit-8 (CCK-8) assay and Transwell assay were used to detect the changes in the viability and migration of the cells. Enzyme-linked immunosorbent assay (ELISA) was used to examine the cytokines IL-4, IL-6, and IL-17, and real-time fluorescence quantitative PCR (RT-qPCR) was performed to detect the expression of circRNA 003353; Western blotting was used to detect the expressions of p-JAK2, pSTAT3, JAK2 and STAT3 proteins in the treated cells.@*RESULTS@#The expression of circRNA 0003353 was significantly increased in PBMCs from RA patients and showed a good performance in assisting the diagnosis of RA (AUC=90.5%, P < 0.001, 95% CI: 0.83-0.98). CircRNA 0003353 expression was positively correlated with ESR, RF and DAS28 (P < 0.05). Treatment with TPL significantly decreased the expression of circRNA 0003353, suppressed the viability and migration ability, decreased the expressions of IL-6 and IL-17, and increased the expression IL-4 in cultured RA-FLS in a time-dependent manner (P < 0.01). TNF-α stimulation of RA-FLS significantly increased the ratios of p-JAK2/JAK2 and p-STAT3/STAT3, which were obviously lowered by TPL treatment (P < 0.01). TPL-treated RA-FLS overexpressing circRNA 0003353 showed significantly increased cell viability and migration ability with decreased IL-4 expression and increased IL-6 and IL-17 expressions and ratios of p-JAK2/ JAK2 and p-STAT3/STAT3 (P < 0.01).@*CONCLUSION@#The expression of circRNA 0003353 is increased in PBMCs in RA patients and in RA-FLS. TPL treatment can regulate JAK2/STAT3 signal pathway and inhibit the inflammatory response and migration of RA-FLS through circRNA 0003353.

Arthritis, Rheumatoid/pathology , Cells, Cultured , Diterpenes/pharmacology , Epoxy Compounds/pharmacology , Fibroblasts/pathology , Humans , Interleukin-17/metabolism , Interleukin-4/metabolism , Interleukin-6/metabolism , Janus Kinase 2/metabolism , Leukocytes, Mononuclear/metabolism , Phenanthrenes/pharmacology , RNA, Circular/metabolism , STAT3 Transcription Factor/metabolism , Signal Transduction/drug effects , Synovial Membrane/pathology
Article in Chinese | WPRIM | ID: wpr-936318


OBJECTIVE@#To explore the expression patterns, prognostic implications, and biological role of leukotriene B4 receptor (LTB4R) in patients with acute myeloid leukemia (AML).@*METHODS@#We collected the data of mRNA expression levels and clinical information of patients with AML from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) database for mRNA expression analyses, survival analyses, Cox regression analyses and correlation analyses using R studio to assess the expression patterns and prognostic value of LTB4R. The correlation of LTB4R expression levels with clinical characteristics of the patients were analyzed using UALCAN. The co-expressed genes LTB4R were screened from Linkedomics and subjected to functional enrichment analysis. A protein-protein interaction network was constructed using STRING. GSEA analyses of the differentially expressed genes (DEGs) were performed based on datasets from TCGA-LAML stratified by LTB4R expression level. We also collected peripheral blood mononuclear cells (PBMCs) from AML patients and healthy donors for examination of the mRNA expression levels of LTB4R and immune checkpoint genes using qRT-PCR. We also examined serum LTB4R protein levels in the patients using ELISA.@*RESULTS@#The mRNA expression level of LTB4R was significantly increased in AML patients (4.898±1.220 vs 2.252±0.215, P < 0.001), and an elevated LTB4R expression level was correlated with a poor overall survival (OS) of the patients (P=0.004, HR=1.74). LTB4R was identified as an independent prognostic factor for OS (P=0.019, HR=1.66) and was associated with FAB subtypes, cytogenetic risk, karyotype abnormalities and NPM1 mutations. The co- expressed genes of LTB4R were enriched in the functional pathways closely associated with AML leukemogenesis, including neutrophil inflammation, lymphocyte activation, signal transduction, and metabolism. The DEGs were enriched in differentiation, activation of immune cells, and cytokine signaling. Examination of the clinical serum samples also demonstrated significantly increased expressions of LTB4R mRNA (P=0.044) and protein (P=0.008) in AML patients, and LTB4R mRNA expression was positively correlated with the expression of the immune checkpoint HAVCR2 (r= 0.466, P=0.040).@*CONCLUSION@#LTB4R can serve as a novel biomarker and independent prognostic indicator of AML and its expression patterns provide insights into the crosstalk of leukemogenesis signaling pathways involving tumor immunity and metabolism.

Humans , Leukemia, Myeloid, Acute/metabolism , Leukocytes, Mononuclear/metabolism , Prognosis , RNA, Messenger/metabolism , Receptors, Leukotriene B4/genetics
Chinese Journal of Epidemiology ; (12): 560-565, 2022.
Article in Chinese | WPRIM | ID: wpr-935427


Objective: To explore the effect and mechanism of activation of peripheral blood mononuclear cell (PBMC) Toll-like receptor (TLR3) signaling pathway in recombinant HBsAg (rHBsAg) immune response. Methods: White blood cells were collected from peripheral blood of 13 healthy donors in the preparation of blood products. PBMC was isolated and treated with Poly I:C (Poly I:C group) and PBS (control group) respectively. 48 h later, some cells were collected and the expressions of TLR3 signaling pathway proteins were detected by flow cytometry. After activating (Poly I:C group)/inactivating (control group) TLR3 signaling pathway, rHBsAg was given to both groups for 72 h, and the proportions of DC, T, B cells and their subsets in PBMC were detected by flow cytometry. Paired t-test, paired samples wilcoxon signed-rank test and canonical correlation analyses were used for statistical analysis. Results: The percentage of TLR3 protein-positive cells (19.21%) and protein expression (8 983.95), NF-κB protein expression (26 193.13), the percentage of pNF-κB protein-positive cells (13.73%) and its proportion in NF-κB (16.03%), and the percentage of pIRF3 protein-positive cells (12.64%) and its proportion in IRF3 (21.80%) in Poly I:C group were higher than those in control group (11.54%, 8 086.00, 22 340.66, 8.72%, 9.71%, 9.57%, 19.12%) (P<0.05), and the percentage of TRIF protein-positive cells (89.75%) and protein expression (304 219.54) were higher in Poly I:C group than in the control group (89.64%, 288 149.72) (P>0.05). After PBMC stimulation by rHBsAg, the proportions of mDC (2.90%), pDC (1.80%), B cell (5.31%) and plasma cell (67.71%) in Poly I:C group were significantly higher than those in the control group (1.83%, 0.81%, 4.23%, 58.82%) (P<0.05). Results of canonical correlation analysis showed that the expression of TLR3 protein was positively correlated with the proportions of plasma cells, the expression of pIRF3 protein was positively correlated with the proportions of plasma cells and mDC, and the percentage of pNF-κB protein-positive cells and the percentage of pIRF3 protein-positive cells were positively correlated with the proportion of CD4+T cells. Conclusions: Poly I:C can activate TLR3/TRIF/NF-κB and TLR3/TRIF/IRF3 signaling pathway, promote the function of downstream signaling molecules, and then promote the maturation of DC, induce the immune responses of CD4+T cell, and promote the maturation and activation of B cells and the immune response of rHBsAg.

Adaptor Proteins, Vesicular Transport/pharmacology , Hepatitis B Surface Antigens , Humans , Immunity , Leukocytes, Mononuclear/metabolism , NF-kappa B , Poly I-C/pharmacology , Signal Transduction , Toll-Like Receptor 3/metabolism , Toll-Like Receptors
Article in Chinese | WPRIM | ID: wpr-939691


AbstractObjective: To investigate the effect of γδ T cells on the proliferation, apoptosis and autophagy of multiple myeloma cells.@*METHODS@#Peripheral blood mononuclear cells (PBMNC) were isolated from healthy volunteers, and stimulated with zoledronic acid (Zol) in combination with rhIL-2. Flow cytometry analysis was used to detected the purity of γδ T cells. γδ T cells were collected and co-cultured with RPMI-8226 or U-266 cells at different effector target ratios. The proliferation of RPMI-8226 or U-266 cell lines were detected by CCK-8. Cell cycle and cell apoptosis were detected by flow cytometry and Western blot.The expressions of autophagy-related proteins were detected by Western blot.@*RESULTS@#γδ T cells can be expanded in vitro. γδ T cells could inhibit the proliferation of RPMI-8226 or U-266 cells, induced cell cycle arrest and promoted apoptosis in an effector target-dependent manner. In addition, γδ T cells could induce autophagy of myeloma cells, inhibited the expression of autophagy-related PI3K, P-AKT and P-mTOR, while increased the expression of AMPK and Beclin-1.@*CONCLUSION@#γδ T cells can inhibit the proliferation of RPMI-8226 and U-266 myeloma cells, induce cell cycle arrest, promote apoptosis, and enhance autophagy in vitro. The mechanism may be related to inhibition of PI3K/AKT/mTOR signaling pathway and/or activation of AMPK/Beclin-1 signaling pathway.

AMP-Activated Protein Kinases/pharmacology , Apoptosis , Autophagy , Beclin-1/pharmacology , Cell Proliferation , Humans , Leukocytes, Mononuclear/metabolism , Multiple Myeloma/metabolism , Phosphatidylinositol 3-Kinases/metabolism , Proto-Oncogene Proteins c-akt/metabolism , T-Lymphocytes , TOR Serine-Threonine Kinases/metabolism
Clinics ; 76: e2432, 2021. tab, graf
Article in English | LILACS | ID: biblio-1153954


OBJECTIVES: Telomeres are a terminal "DNA cap" that prevent chromosomal fusion and degradation. However, aging is inherent to life, and so is the loss of terminal sequences. Telomerase is a specialized reverse transcriptase encoded by self-splicing introns that counteract chromosome erosion. Telomerase activity is observed during early embryonic development, but after the blastocyst stage, the expression of telomerase reduces. The consequences of either insufficient or unrestrained telomerase activity underscore the importance of ongoing studies aimed at elucidating the regulation of telomerase activity in humans. In the present study, we aimed to standardize a simplified telomerase repeat-amplification protocol (TRAP) assay to detect telomerase activity in unstimulated and PHA-stimulated mononuclear cells. METHODS and RESULTS: Our optimized qPCR-based can efficiently evaluate telomerase activity. Quantification of protein and DNA between unstimulated and PHA-stimulated peripheral blood mononuclear cells revealed cellular activation and cell-cycle entry. The assay also showed that relative telomerase activity is significantly different between these two conditions, supporting the applicability of the assay. Furthermore, our findings corroborated that telomerase activity decreases with age. CONCLUSIONS: Telomeres and telomerase are implicated in aging and development of chronic diseases and cancer; however, difficulty in accessing commercial kits to investigate these aspects is a critical constraint in health surveillance studies. Our optimized assay was successfully used to differentiate telomerase activity between unstimulated and stimulated cells, clearly showing the reactivation of telomerase upon cell activation. This assay is affordable, reproducible, and can be executed in resource-limited settings.

Humans , Female , Pregnancy , Telomerase/genetics , Telomerase/metabolism , Neoplasms , Aging , Leukocytes, Mononuclear/metabolism , Chronic Disease , Cost-Benefit Analysis
J. appl. oral sci ; 27: e20180529, 2019. graf
Article in English | LILACS, BBO | ID: biblio-1012507


Abstract Objectives: Dental composites release unreacted resin monomers into the oral environment, even after polymerization. Periodontal cells are, therefore, exposed to substances that potentially elicit the immune inflammatory response. The underlying molecular mechanisms associated with the interaction between resin monomers and human immune cells found in the gingival crevicular fluid are not fully understood yet. This study investigated the ability of bisphenol A-glycidyl methacrylate (BISGMA), urethane dimethacrylate (UDMA) and triethylene glycol dimethacrylate (TEGDMA) to induce apoptosis and cytokine release by human leukocytes stimulated with a periodontal pathogen. Methodology: Peripheral blood mononuclear cells (PBMC) from 16 healthy individuals were included in this study. To determine the toxicity, the PBMC were incubated for 20 hours, with monomers, for the analysis of cell viability using MTT assay. To evaluate cell death in the populations of monocytes and lymphocytes, they were exposed to sub-lethal doses of each monomer and of heat-inactivated Porphyromonas gingivalis (P. gingivalis) for 5 hours. Secretions of IL-1β, IL-6, IL-10 and TNF-α were determined by ELISA after 20 hours. Results: UDMA and TEGDMA induced apoptosis after a short-time exposure. Bacterial challenge induced significant production of IL-1β and TNF-α (p<0.05). TEGDMA reduced the bacterial-induced release of IL-1β and TNF-α, whereas UDMA reduced IL-1β release (p<0.05). These monomers did not affect IL-10 and IL-6 secretion. BISGMA did not significantly interfere in cytokine release. Conclusions: These results show that resin monomers are toxic to PBMC in a dose-dependent manner, and may influence the local immune inflammatory response and tissue damage mechanisms via regulation of bacterial-induced IL-1β and TNF-α secretion by PBMC.

Humans , Polyethylene Glycols/pharmacology , Polymethacrylic Acids/pharmacology , Polyurethanes/pharmacology , Leukocytes, Mononuclear/drug effects , Cytokines/metabolism , Bisphenol A-Glycidyl Methacrylate/pharmacology , Porphyromonas gingivalis/physiology , Methacrylates/pharmacology , Reference Values , Time Factors , Enzyme-Linked Immunosorbent Assay , Leukocytes, Mononuclear/metabolism , Cell Survival/drug effects , Reproducibility of Results , Analysis of Variance , Cytokines/analysis , Cytokines/drug effects , Apoptosis/drug effects , Statistics, Nonparametric , Necrosis
J. bras. nefrol ; 40(4): 333-338, Out.-Dec. 2018. tab, graf
Article in English | LILACS | ID: biblio-984593


ABSTRACT Introduction: Fabry disease (FD) is a disorder caused by mutations in the gene encoding for lysosomal enzyme α-galactosidase A (α-GAL). Reduced α-GAL activity leads to progressive accumulation of globotriaosylceramide (Gb3), also known as CD77. The recent report of increased expression of CD77 in blood cells of patients with FD indicated that this molecule can be used as a potential marker for monitoring enzyme replacement therapy (ERT). Objective: The purpose of this study was to evaluate the CD77 levels throughout ERT in FD patients (V269M mutation). Methods: We evaluated the fluctuations in PBMC (peripheral blood mononuclear cell) membrane CD77 expression in FD patients undergoing ERT and correlated these levels with those observed in different cell types. Results: A greater CD77 expression was found in phagocytes of patients compared to controls at baseline. Interestingly, the variability in CD77 levels is larger in patients at baseline (340 - 1619 MIF) and after 12 months of ERT (240 - 530 MIF) compared with the control group (131 - 331 MFI). Furthermore, by analyzing the levels of CD77 in phagocytes from patients throughout ERT, we found a constant decrease in CD77 levels. Conclusion: The increased CD77 levels in the phagocytes of Fabry carriers together with the decrease in CD77 levels throughout ERT suggest that measuring CD77 levels in phagocytes is a promising tool for monitoring the response to ERT in FD.

RESUMO Introdução: A doença de Fabry (DF) é um distúrbio causado por mutações no gene que codifica a enzima lisossômica α-galactosidase A (α-GAL). A redução da atividade de α-GAL leva ao acúmulo progressivo de globotriaosilceramida (Gb3), também conhecida como CD77. O recente relato de aumento da expressão de CD77 em células sanguíneas de pacientes com DF indicou que essa molécula pode ser utilizada como um potencial marcador para o monitoramento da terapia de reposição enzimática (TRE). Objetivo: O objetivo deste estudo foi avaliar os níveis de CD77 ao longo da TRE em pacientes com DF (mutação V269M). Métodos: Foram avaliadas as flutuações na expressão de CD77 nas membranas das CMSP (células mononucleares do sangue periférico) em pacientes com DF submetidos à TRE e correlacionados com aqueles observados em diferentes tipos de células. Resultados: Uma maior expressão de CD77 foi encontrada em fagócitos de pacientes em comparação aos controles no início do estudo. Curiosamente, a variabilidade nos níveis de CD77 é maior em pacientes no início do estudo (340 - 1619 MIF) e após 12 meses de TRE (240 - 530 MIF) em comparação com o grupo controle (131 - 331 MFI). Além disso, analisando os níveis de CD77 em fagócitos de pacientes ao longo da TRE, encontramos uma diminuição constante nos níveis de CD77. Conclusão: O aumento nos níveis de CD77 nos fagócitos de portadores de Fabry, juntamente com a diminuição nos níveis de CD77 ao longo da TRE, sugerem que medir os níveis de CD77 nos fagócitos é uma ferramenta promissora para monitorar a resposta à TRE na DF.

Humans , Male , Female , Adult , Young Adult , Trihexosylceramides/biosynthesis , Leukocytes, Mononuclear/metabolism , Fabry Disease/drug therapy , Fabry Disease/blood , alpha-Galactosidase/therapeutic use , Enzyme Replacement Therapy , Trihexosylceramides/analysis , Leukocytes, Mononuclear/chemistry
Braz. j. med. biol. res ; 51(8): e7334, 2018. graf
Article in English | LILACS | ID: biblio-951739


Pregnancy-induced hypertension (PIH) causes significant maternal and fetal morbidity and mortality. A decreased number of regulatory T (Treg) cells is associated with the pathogenesis of PIH. The programmed cell death-1 (PD-1)/PD-ligand 1 (PD-L1) pathway is critical to normal pregnancy (NP) by promoting Treg cell development. However, the relationship between PD-1/PD-L1 and Treg differentiation in PIH has not been fully elucidated. In this study, venous blood was obtained from 20 NP and 58 PIH patients. Peripheral blood mononuclear cells (PBMCs) were isolated from venous blood. The levels of Treg-related cytokines (TGF-β, IL-10, and IL-35) in serum and PBMCs were measured by ELISA. The percentage of Treg cells in PBMCs was assessed by flow cytometry. The mRNA levels of Treg-specific transcription factor Foxp3 in PBMCs, and PD-1 and PD-L1 in Treg cells were detected by qRT-PCR. The protein levels of PD-1 and PD-L1 in Treg cells were evaluated by western blot. The serum levels of TGF-β, IL-10, IL-35, and Foxp3 mRNA expression and CD4+CD25+ Treg cell percentage in PBMCs were decreased in PIH. Furthermore, a significant increase of PD-1 in Treg cells was found in PIH compared with NP. In addition, PD-L1 Fc, an activator of PD-1/PD-L1 pathway, increased Treg cell percentage, enhanced Foxp3 mRNA expression, and elevated levels of TGF-β, IL-10, and IL-35 in PBMCs. However, anti-PD-L1 mAb exerted a reverse effect. These findings revealed that PD-L1 Fc had a favorable effect on Treg cell differentiation, indicating a potential therapeutic value of PD-1/PD-L1 pathway for PIH treatment.

Humans , Female , Pregnancy , Leukocytes, Mononuclear/chemistry , Interleukins/metabolism , Interleukin-10/metabolism , Apoptosis , Hypertension, Pregnancy-Induced/metabolism , B7-H1 Antigen/metabolism , Enzyme-Linked Immunosorbent Assay , Leukocytes, Mononuclear/metabolism , Case-Control Studies , Blotting, Western , Transforming Growth Factor beta/metabolism , T-Lymphocytes, Regulatory/metabolism , Real-Time Polymerase Chain Reaction
Braz. j. med. biol. res ; 50(10): e6139, 2017. tab, graf
Article in English | LILACS | ID: biblio-888929


Augmenter of liver regeneration (ALR) is a thermostable cytokine that was originally identified to promote the growth of hepatocytes. This study was conducted to explore the expression and function of ALR in multiple myeloma (MM), a common hematologic malignancy. Real-time PCR and western blot analysis were performed to detect the expression of ALR in U266 human MM cells and healthy peripheral blood mononuclear cells (PBMCs). U266 MM cells were exposed to 20 or 40 μg/mL of recombinant ALR and tested for cell proliferation. Small interfering RNA-mediated silencing of ALR was done to investigate the role of ALR in cell proliferation, apoptosis, and cytokine production. Compared to PBMCs, U266 MM cells exhibited significantly higher levels of ALR at both the mRNA and protein levels. The addition of recombinant ALR protein significantly promoted the proliferation of U266 cells. In contrast, knockdown of ALR led to a significant decline in the viability and proliferation of U266 cells. Annexin-V/PI staining analysis demonstrated that ALR downregulation increased apoptosis in U266 MM cells, compared to control cells (20.1±1.1 vs 9.1±0.3%, P<0.05). Moreover, ALR depletion reduced the Bcl-2 mRNA level by 40% and raised the Bax mRNA level by 2-fold. Additionally, conditioned medium from ALR-depleted U266 cells had significantly lower concentrations of interleukin-6 than control cells (P<0.05). Taken together, ALR contributed to the proliferation and survival of U266 MM cells, and targeting ALR may have therapeutic potential in the treatment of MM.

Humans , Apoptosis/drug effects , Cell Proliferation/drug effects , Multiple Myeloma/metabolism , Proteins/pharmacology , Blotting, Western , Cell Line, Tumor , Cytokines/biosynthesis , Down-Regulation , Flow Cytometry , Leukocytes, Mononuclear/metabolism , Multiple Myeloma/immunology , Proteins/physiology , Real-Time Polymerase Chain Reaction , Recombinant Proteins/pharmacology , RNA, Small Interfering/metabolism
Braz. j. med. biol. res ; 50(8): e5163, 2017. graf
Article in English | LILACS | ID: biblio-888986


Pseudobrickellia brasiliensis (Asteraceae) is a plant commonly known as arnica-do-campo and belongs to the native flora of the Brazilian Cerrado. The alcoholic extract of the plant has been used as an anti-inflammatory agent in folk medicine, but the biological mechanism of action has not been elucidated. The present study evaluated the composition of P. brasiliensis aqueous extract and its effects on pro-inflammatory cytokine production and lymphocyte proliferation. The extracts were prepared by sequential maceration of P. brasiliensis leaves in ethanol, ethyl acetate, and water. Extract cytotoxicity was evaluated by trypan blue exclusion assay, and apoptosis and necrosis were measured by staining with annexin V-FITC and propidium iodide. The ethanolic (ETA) and acetate (ACE) extracts showed cytotoxic effects. The aqueous extract (AQU) was not cytotoxic. Peripheral blood mononuclear cells stimulated with phorbol myristate acetate and ionomycin and treated with AQU (100 μg/mL) showed reduced interferon (IFN)-γ and tumor necrosis factor (TNF)-α expression. AQU also inhibited lymphocyte proliferative response after nonspecific stimulation with phytohemagglutinin. The aqueous extract was analyzed by liquid chromatography coupled with photodiode array detection and mass spectrometry. Quinic acid and its derivatives 5-caffeoylquinic acid and 3,5-dicaffeoylquinic acid, as well as the flavonoids luteolin and luteolin dihexoside, were detected. All these compounds are known to exhibit anti-inflammatory activity. Taken together, these findings demonstrate that P. brasiliensis aqueous extract can inhibit the pro-inflammatory cytokine production and proliferative response of lymphocytes. These effects may be related to the presence of chemical substances with anti-inflammatory actions previously reported in scientific literature.

Humans , Anti-Inflammatory Agents/pharmacology , Asteraceae/chemistry , Cell Proliferation/drug effects , Interferon-gamma/drug effects , Lymphocytes/drug effects , Plant Extracts/pharmacology , Tumor Necrosis Factor-alpha/drug effects , Cell Survival/drug effects , Chromatography, High Pressure Liquid , Interferon-gamma/metabolism , Leukocytes, Mononuclear/drug effects , Leukocytes, Mononuclear/metabolism , Plant Extracts/chemistry , Time Factors , Tumor Necrosis Factor-alpha/metabolism
Braz. j. med. biol. res ; 49(3): e4853, Mar. 2016. tab, graf
Article in English | LILACS | ID: lil-771933


The objective of this study was to examine the relationship between the expression of B cell activating factor (BAFF) and BAFF receptor in patients with disease activity of systemic lupus erythematosus (SLE). Real-time RT-PCR was used to examine BAFF mRNA expression in peripheral blood monocytes of active and stable SLE patients and healthy controls. The percentage of BAFF receptor 3 (BR3) on B lymphocytes was measured by flow cytometry. Soluble BAFF levels in serum were assayed by ELISA. Microalbumin levels were assayed by an automatic immune analysis machine. BAFF mRNA and soluble BAFF levels were highest in the active SLE group, followed by the stable SLE group, and controls (P<0.01). The percentage of BR3 on B lymphocytes was downregulated in the active SLE group compared with the stable SLE group and controls (P<0.01). BAFF mRNA levels and soluble BAFF levels were higher in patients who were positive for proteinuria than in those who were negative (P<0.01). The percentage of BR3 on B lymphocytes was lower in patients who were positive for proteinuria than in those who were negative (P<0.01). The BAFF/BR3 axis may be over-activated in SLE patients. BAFF and BR3 levels may be useful parameters for evaluating treatment.

Humans , Male , Female , Adolescent , Adult , Middle Aged , Young Adult , B-Cell Activating Factor/metabolism , B-Cell Activation Factor Receptor/metabolism , Lupus Erythematosus, Systemic/metabolism , Albuminuria/urine , B-Cell Activating Factor/analysis , B-Cell Activating Factor/genetics , B-Cell Activation Factor Receptor/analysis , B-Cell Activation Factor Receptor/genetics , B-Lymphocytes/metabolism , Biomarkers/metabolism , Leukocytes, Mononuclear/metabolism , Lupus Erythematosus, Systemic/immunology , RNA, Messenger/genetics , RNA, Messenger/metabolism
Arch. argent. pediatr ; 113(5): 404-410, oct. 2015. tab
Article in Spanish | LILACS | ID: lil-757061


Introducción. Los trastornos crónicos (TC) en etapas tempranas de la vida pueden influir en diferentes dimensiones de la calidad de vida relacionada con la salud (CVRS) de los niños/as. Objetivo. Comparar la CVRS de niños/as con TC confirmados, con TC declarados y sin TC. Población y método. Estudio transversal en el marco de una investigación mayor realizada en escuelas de Córdoba y Bahía Blanca, y en los hospitales Italiano y Garrahan de Buenos Aires, Argentina, en 2012. La presencia de TC fue establecida por diagnóstico médico en hospitales o por declaración de sus cuidadores en escolares. Los niños/as de 8 a 12 años respondieron el cuestionario KIDSCREEN-52 sobre CVRS, una escala de desarrollo puberal y una escala de recursos económicos familiares. Los cuidadores indicaron el nivel educativo materno. Se estimó la asociación entre TC y CVRS ajustada por sexo, edad, desarrollo puberal, nivel educativo materno y nivel socioeconómico. Resultados. Participaron 670 duplas niños/as-cuidadores, 13,3% (n= 89) con TC confirmados (muestras de hospitales), 14,5% (n= 97) escolares con trastornos declarados y el resto eran escolares sanos. La edad promedio fue 10,2 años (desvío estándar= 1,01); 54,8% fueron niñas. Tener TC confirmados se asoció a una mayor frecuencia de bajo bienestar físico (OR 2,61; IC 95%: 1,43-4,76), mientras que la presencia de TC declarados se asoció con bajas puntuaciones en bienestar psicológico (OR 1,96; IC 95%: 1,063,63), autopercepción (OR 2,22; IC 95%: 1,283,87) y relación con los padres (OR 2,04; IC 95%: 1,21-3,44). Conclusiones. Los niños/as con TC confirmados mostraron con mayor frecuencia malestar físico y los que tenían TC declarados manifestaron malestar en áreas psicosociales, en comparación con los niños/as sin trastorno.

Introduction. Chronic conditions (CCs) in the early stages of life may have an impact on various dimensions of health-related quality of life (HRQoL) in children. Objective. To compare HRQoL in children with confirmed CCs, reported CCs, and without CC. Population and Method. Cross-sectional study conducted in 2012 in the context of a larger research study carried out at schools in Córdoba and Bahía Blanca, and at Hospital Italiano of Buenos Aires and Hospital Prof. Dr. Juan P. Garrahan at Buenos Aires. The presence of a chronic condition was established by medical diagnosis at the hospital or as reported by schoolchildren's caregivers. Eight-to-twelve year-old children completed the KIDSCREEN-52 questionnaire on HRQoL, a pubertal development scale, and a family financial resource scale. The association between CCs and HRQoL adjusted by sex, age, pubertal development, maternal education level, and socioeconomic level was estimated. Results. Six hundred and seventy children/ caregiver dyads participated; 13.3% (n= 89) had confirmed CCs, 14.5% (n= 97) were schoolchildren with reported CCs, and the rest corresponded to healthy schoolchildren. Their average age was 10.2 years old (standard deviation= 1.01); 54.8% were girls. Having a confirmed CC was associated with a higher frequency of low physical wellbeing (odds ratio --OR--: 2.61; 95% confidence interval --95% CI--:1.43-4.76), while the presence of a reported CC was associated with a low score in psychological well-being (OR: 1.96; 95% CI: 1.06-3.63), self-perception (OR: 2.22; 95% CI: 1.28-3.87), and parent relations (OR: 2.04; 95% CI: 1.21-3.44). Conclusions. Children with confirmed CCs showed a higher frequency of physical discomfort, and those with reported CCs showed discomfort in psychosocial areas compared to children without CCs.

Humans , Cytokines/genetics , Cytokines/metabolism , Proteome , Transcriptome , Cluster Analysis , Computational Biology , Gene Expression Profiling , Leukocytes, Mononuclear/metabolism , Proteomics
Arch. argent. pediatr ; 113(5): 411-418, oct. 2015. graf, tab
Article in Spanish | LILACS | ID: lil-757062


Introducción. Existen evidencias de la asociación de determinantes sociales con la salud infantil. Objetivo. Identificar características sociodemográficas asociadas a desigualdades en la salud infantil y evaluar el efecto acumulado sobre la salud de factores de riesgo basados en estas características. Población y métodos. Evaluamos niños de 4-13 años, de Bariloche, entre junio de 2008 y mayo de 2009. Características sociodemográficas consideradas: nivel socioeconómico, educación materna, embarazo adolescente, cobertura médica, inseguridad y hábitos familiares. Valoramos la percepción parental de la salud física y socioemocional, el estado nutricional y la salud bucal en relación con dichas características y con la acumulación de factores de riesgo. Utilizamos encuesta, antropometría y examen bucal. Resultados. Participaron 180 escolares. El nivel educativo materno se asoció con la salud física, socioemocional y bucal del niño. El porcentaje de niños con piezas faltantes o caries fue 77% entre aquellos cuyas madres, como máximo, habían completado el primario, comparado con 13% entre aquellos cuyas madres habían completado estudios terciarios/universitarios. La posibilidad de percepción de salud física y socioemocional no óptima aumentó con cada factor de riesgo 1,8 y 1,4 veces, respectivamente, y la posibilidad de caries o piezas faltantes se duplicó con cada factor de riesgo adicional. El 27,3% de los escolares presentó sobrepeso y el 8,7%, obesidad, y no se encontró asociación con características sociodemográficas. Conclusiones. El bajo nivel socioeconómico familiar y educativo materno se asoció con una mayor prevalencia de resultados de salud desfavorables. Múltiples factores de riesgo tienen un efecto acumulado sobre la percepción parental de la salud física y socioemocional y la salud bucal.

Introduction. There is evidence of an association between social determinants and child health. Objective. To identify sociodemographic characteristics related to child health inequalities and to analize the cumulative effect on health of risk factors based on these characteristics. Population and Methods. We evaluated 4-13 year-old children in Bariloche between June 2008 and May 2009. The following sociodemographic characteristics were taken into account: socioeconomic level, maternal education, adolescent pregnancy, medical coverage, unsafeness, and family habits. We assessed parental perception of physical, and social and emotional health, nutritional status and oral health in relation to these characteristics and the accumulation of risk factors. We used survey, anthropometry and oral examination. Results. One hundred and eighty students participated. The level of maternal education was associated with the child's physical, social and emotional, and oral health. The percentage of children with missing teeth or cavities reached 77% among those whose mothers had, at most, completed primary school, compared to 13% among those whose mothers had completed tertiary school or university. The possibility of perceiving a non-optimal physical, and social and emotional health increased 1.8 and 1.4 times with each risk factor, respectively, and the possibility of having missing teeth or cavities was twice as much with each additional risk factor. Overweight and obesity was observed in 27.3% and 8.7% of students, respectively, and no relationship was found with sociodemographic characteristics. Conclusions. A low family socioeconomic level and a low maternal education level were associated with a higher prevalence of unfavorable health outcomes. Multiple risk factors have an cumulative effect on parental perception of physical, social and emotional, and oral health.

Humans , Cell Transformation, Viral/genetics , Gene Expression Profiling , Transcriptome , Antigens, Surface/genetics , Antigens, Surface/metabolism , Cell Line, Transformed , Cell Proliferation , Cells, Cultured , Flow Cytometry , Gene Expression , Genes, Viral , Genotype , /genetics , Leukocytes, Mononuclear/metabolism , Leukocytes, Mononuclear/virology , Transcription, Genetic , Virus Latency
Mem. Inst. Oswaldo Cruz ; 110(6): 781-785, Sept. 2015. tab, graf
Article in English | LILACS | ID: lil-763092


Paracoccidioidomycosis (PCM) is caused by dimorphic fungi from theParacoccidioides brasiliensis complex. Previous studies have demonstrated that the severity of disease is associated with a T-helper 2 immune response characterised by high interleukin (IL)-4 production. In the present study we analysed two polymorphisms in the IL-4gene (-590 C/T and intron-3 microsatellite) in 76 patients with PCM and 73 control subjects from an endemic area. The production of IL-4 by peripheral blood mononuclear cells after antigen or phytohaemagglutinin stimulation was determined by ELISA. A significant correlation was observed between the RP2/RP2 intron-3 genotype and infection with Paracoccidioides sp.(p = 0.011), whereas the RP1/RP1 genotype was correlated with resistance. No significant correlation was observed for the IL-4promoter polymorphism. Furthermore, the low IL-4 expression observed in the control group compared with patients was associated with the RP1/RP1 genotype. These results suggest that IL-4polymorphisms might be associated with the ability of the host to control Paracoccidioides sp.infection. The relevance of this polymorphism is supported by the observation that patients with disease produce high levels of IL-4 following mitogen or antigen stimulation. The IL-4gene is located in the cytokine cluster region of chromosome 5 where other polymorphisms have also been described.

Adult , Female , Humans , Male , Middle Aged , Young Adult , Endemic Diseases , Genetic Predisposition to Disease , /genetics , /metabolism , Paracoccidioidomycosis/immunology , Polymorphism, Genetic/immunology , Enzyme-Linked Immunosorbent Assay , Leukocytes, Mononuclear/immunology , Leukocytes, Mononuclear/metabolism , Microsatellite Repeats , Paracoccidioidomycosis/epidemiology , Promoter Regions, Genetic/genetics , Statistics, Nonparametric
Rev. latinoam. enferm. (Online) ; 23(4): 642-650, July-Aug. 2015. tab
Article in English | LILACS, BDENF | ID: lil-761700


AbstractObjectives: develop and validate the content of a tool about nursing care production.Method: the data were collected between 2011 and 2013, based on focus groups, the application of semistructured questionnaires (prototype test) and the Delphi technique. The focus groups were used to produce the instrument items and held at three hospitals in the interior of the State of São Paulo, involving 20 nurses. A panel of 10 experts evaluated the instrument.Results: after two phases of the Delphi technique, the tool consisted of eight items. The content validity index of the scale corresponded to ≥0.9 and the content validity of the items ranged between 0.8 and 1.0, indicating the maintenance of the structure and content. The assertion on the applicability in daily nursing practice showed a content validity index of the scale equal to 0.8.Conclusion: this study permitted the development and content validation of scale on nursing care production, equipping the nurses in their management practice.

ResumoObjetivos:desenvolver e validar o conteúdo de um instrumento sobre produção do cuidado de enfermagem.Método:a coleta de dados ocorreu entre 2011 e 2013, a partir de grupos focais, aplicação de questionários semiestruturados (teste do protótipo) e técnica Delphi. Os grupos focais foram utilizados para geração de itens do instrumento e realizados em três hospitais do interior do Estado de São Paulo, com a participação de 20 enfermeiros. A apreciação do instrumento foi conduzida por um painel de 10 especialistas.Resultados:após duas fases da técnica Delphi, o instrumento passou a ser constituído por oito itens. O índice de validade do conteúdo da escala foi de ≥0,9 e a validade dos conteúdos dos itens apresentou variação de 0,8 a 1,0, indicando a manutenção da estrutura e do conteúdo. A afirmativa referente à aplicabilidade na prática diária do enfermeiro apresentou índice de validade do conteúdo da escala de 0,8.Conclusão:este estudo possibilitou desenvolver e validar o conteúdo de uma escala sobre produção do cuidado de enfermagem, instrumentalizando os enfermeiros em sua prática gerencial.

ResumenObjetivos:desarrollar y validar el contenido de un instrumento sobre producción del cuidado de enfermería.Método:los datos fueron recolectados entre 2011 y 2013, a partir de grupos focales, aplicación de cuestionarios semiestructurados (prueba del prototipo) y técnica Delphi. Los grupos focales fueron utilizados para generar ítems del instrumento y organizados en tres hospitales del interior del Estado de São Paulo, con la participación de 20 enfermeros. La apreciación del instrumento fue conducida por un panel de 10 especialistas.Resultados:tras dos fases de la técnica Delphi, el instrumento pasó a ser constituido por ocho ítems. El índice de validez de contenido de la escala fue ≥0,9 y la validez de los contenidos mostró variación de 0,8 a 1,0, indicando la manutención de la estructura y del contenido. La afirmativa respecto a la aplicabilidad en la práctica diaria del enfermero mostró índice de validez del contenido de la escala de 0,8.Conclusión:este estudio permitió desarrollar y validar el contenido de una escala sobre producción del cuidado de enfermería, instrumentalizando los enfermeros en su práctica gerencial.

Humans , Male , Female , Adult , Middle Aged , Aged , /blood , Gene Expression Regulation , Graft Rejection/blood , Kidney Transplantation , Leukocytes, Mononuclear/metabolism , Age Factors , /immunology , Graft Rejection/immunology , Graft Rejection/pathology , Leukocytes, Mononuclear/immunology , Leukocytes, Mononuclear/pathology , Postoperative Period , Time Factors
Rev. chil. pediatr ; 86(2): 103-111, abr. 2015. graf, tab
Article in Spanish | LILACS | ID: lil-752887


Introducción: Desnutrición, retardo en el crecimiento e infecciones oportunistas sobrevienen a alteraciones metabólicas, inmunológicas y gastrointestinales que produce el virus de la inmunodeficiencia humana (VIH). La deficiencia de zinc se ha asociado con deterioro nutricional, falla en el crecimiento y riesgo de infecciones. El objetivo de este estudio fue asociar los niveles de zinc en células mononucleares de sangre periférica (PBMC) con el estado nutricional en niños infectados por el VIH y en niños no infectados expuestos al virus. Pacientes y Método: Estudio analítico observacional, transversal, en 17 niños infectados y 17 expuestos, entre 2 y 10 años de edad. Se realizó valoración antropométrica, historia clínica-nutricional, recordatorio de 24 horas, medición de actividad física y determinación de zinc en PBMC por citometría de fiujo. Resultados: La talla para la edad, el consumo de energía, y la adecuación de energía, proteínas y zinc alimentario fueron significativamente mayores en los niños expuestos comparados con los niños infectados (p < 0,05). No se hallaron diferencias significativas en el índice de masa corporal, los niveles de zinc en monocitos, linfocitos CD4+ y CD4- entre los dos grupos de estudio (p > 0,05); sin embargo, la mediana de los niveles de zinc en monocitos de pacientes infectados fue mayor (218,6) comparado con el grupo control (217,0). No se encontró asociación entre consumo de zinc y niveles de zinc intracelular. Conclusiones: El deterioro del estado nutricional y el retardo en el crecimiento en niños estuvo asociado al VIH, pero no a los niveles de zinc intracelular. El consumo alimentario de este nutriente no se asoció a niveles de zinc en monocitos y linfocitos CD4+ y CD4-.

Introduction: Malnutrition, growth retardation and opportunistic infections outlast the metabolic, immune and gastrointestinal disorders produced by HIV. Zinc deficiency has been associated with deteriorating nutritional status, growth failure, and risk of infection. The aim of this study is to determine the association between zinc levels in peripheral blood mononuclear cells (PBMC) and the nutritional status of HIV-infected and uninfected children exposed to the virus. Patients and Methods: An analytical, observational, cross-sectional study was conducted on 17 infected and 17 exposed children, aged 2-10 years. Anthropometric measurements, clinical and nutritional history, 24 h recall, measurement of physical activity, and zinc in PBMC by fiow cytometry analysis were recorded. Results: Height according to age, energy consumption and adequacy of energy, protein and dietary zinc were significantly higher in children exposed to the virus compared to those infected with HIV (P < 0.05). No significant differences were found in BMI, levels of zinc in monocytes, CD4+ and CD4- lymphocytes between the two study groups (P > 0.05). However, the median levels of zinc in monocytes of infected patients was higher (218.6) compared to the control group (217.0). No association was found between zinc intake and levels of intracellular zinc. Conclusions: The deterioration of nutritional status and growth retardation in children were associated with HIV, but not with the levels of intracellular zinc. The dietary intake of this nutrient was not associated with levels of zinc in monocytes or CD4+ and CD4- lymphocytes.

Humans , Child, Preschool , Child , Zinc/metabolism , Leukocytes, Mononuclear/metabolism , HIV Infections/complications , Nutritional Status , Zinc/administration & dosage , Lymphocytes/metabolism , Monocytes/metabolism , CD4-Positive T-Lymphocytes , Cross-Sectional Studies , Flow Cytometry
Clinics ; 70(2): 120-125, 2/2015. tab, graf
Article in English | LILACS | ID: lil-741425


OBJECTIVES: To explore the microendoscopic discectomy technique and inclusion criteria for the treatment of recurrent lumbar disc herniation and to supply feasible criteria and technical notes to avoid complications and to increase the therapeutic effect. METHODS: A consecutive series of 25 patients who underwent posterior microendoscopic discectomy for recurrent lumbar disc herniation were included. The inclusion criteria were as follows: no severe pain in the lumbar region, no lumbar instability observed by flexion-extension radiography and no intervertebral discitis or endplate damage observed by magnetic resonance imaging. All patients were diagnosed by clinical manifestations and imaging examinations. RESULTS: Follow-up visits were carried out in all cases. Complications, such as nerve injuries, were not observed. The follow-up outcomes were graded using the MacNab criteria. A grade of excellent was given to 12 patients, good to 12 patients and fair to 1 patient. A grade of excellent or good occurred in 96% of cases. One patient relapsed 3 months after surgery and then underwent lumbar interbody fusion and inner fixation. The numerical rating scale of preoperative leg pain was 7.4± 1.5, whereas it decreased to 2.1±0.8 at 7 days after surgery. The preoperative Oswestry disability index of lumbar function was 57.5±10.0, whereas it was 26.0±8.5 at 7 days after surgery. CONCLUSION: In these cases, microendoscopic discectomy was able to achieve satisfactory clinical results. Furthermore, it has advantages over other methods because of its smaller incision, reduced bleeding and more efficient recovery. .

Humans , Centrifugation/methods , Leukocytes, Mononuclear/metabolism , Transfection/methods , Cell Survival/physiology , Leukocytes, Mononuclear/cytology , RNA Interference/physiology , RNA, Small Interfering/genetics
Gut and Liver ; : 370-380, 2015.
Article in English | WPRIM | ID: wpr-203889


BACKGROUND/AIMS: This study investigated the expression of T cell immunoglobulin- and mucin-domain-containing molecule 3 (TIM-3), human beta-defensin (HBD)-2, forkhead box protein 3 (FOXP3), and the frequency of CD4+ CD25+ FOXP3+ regulatory T cells (Tregs) in children with Crohn's disease (CD) during infliximab therapy. METHODS: We enrolled 20 CD patients who received infliximab treatment for 1 year. Peripheral blood and colonic mucosal specimens were collected from all CD patients and from healthy control individuals. RESULTS: A significant difference in TIM-3 mRNA expression was evident in peripheral blood mononuclear cells and colonic mucosa between CD patients before infliximab therapy and the healthy controls (p<0.001 and p=0.005, respectively). A significant difference in HBD-2 mRNA expression was found in colonic mucosa between CD patients before infliximab therapy and the healthy controls (p=0.013). In the active phase of CD, at baseline, the median percentage of T cells that were CD25+ FOXP3+ was 1.5% (range, 0.32% to 3.49%), which increased after inflixmab treatment for 1 year to 2.2% (range, 0.54% to 5.02%) (p=0.008). CONCLUSIONS: Our study suggests that both the adaptive and innate immune systems are closely linked to each other in CD pathogenesis. And the results of our study indicate that it could be a useful therapeutic tool, where restoration of TIM-3, HBD-2 and the function of Tregs may repair the dysfunctional immunoregulation in CD.

Adolescent , Case-Control Studies , Colon/immunology , Crohn Disease/drug therapy , Female , Forkhead Transcription Factors/metabolism , Gastrointestinal Agents/therapeutic use , Humans , Infliximab/therapeutic use , Intestinal Mucosa/immunology , Leukocytes, Mononuclear/metabolism , Male , Membrane Proteins/metabolism , T-Lymphocytes, Regulatory/immunology , beta-Defensins/metabolism