RESUMEN
Malaria remains one of the leading causes of death in sub-Saharan Africa, ranked in the top three infectious diseases in the world. Plants of the Eriosema genus have been reported to be used for the treatment of this disease, but scientific evidence is still missing for some of them. In the present study, the in vitro antiplasmodial activity of the crude extract and compounds from Eriosema montanum Baker f. roots were tested against the 3D7 strain of Plasmodium falciparum and revealed using the SYBR Green, a DNA intercalating compound. The cytotoxicity effect of the compounds on a human cancer cell line (THP-1) was assessed to determine their selectivity index. It was found that the crude extract of the plant displayed a significant antiplasmodial activity with an IC50 (µg/mL) = 17.68 ± 4.030 and a cytotoxic activity with a CC50 (µg/mL) = 101.5 ± 12.6, corresponding to a selective antiplasmodial activity of 5.7. Bioactivity-guided isolation of the major compounds of the roots' crude extract afforded seven compounds, including genistein, genistin and eucomic acid. Under our experimental conditions, using Artemisinin as a positive control, eucomic acid showed the best inhibitory activity against the P. falciparum 3D7, a well-known chloroquine-sensitive strain. The present results provide a referential basis to support the traditional use of Eriosema species in the treatment of malaria.
Asunto(s)
Antimaláricos/farmacología , Fabaceae/química , Raíces de Plantas/química , Plasmodium falciparum/efectos de los fármacos , Antimaláricos/química , Antimaláricos/aislamiento & purificación , Muerte Celular/efectos de los fármacos , Cloroquina/farmacología , Mezclas Complejas , Humanos , Células THP-1RESUMEN
The activation of NOD-, LRR-, and pyrin domain-containing protein 3 (NLRP3) inflammasome and/or its components is associated with the physio-pathogenesis of many respiratory diseases including asthma, COPD (chronic obstructive pulmonary disease), SARS Cov-2 (severe acute respiratory syndrome coronavirus 2), and in several autoimmune diseases. Hibiscus noldeae Baker f. has been widely reported to be traditionally used in the treatment of different ailments, some of which are of inflammatory background such as asthma, wounds, headache, etc. However, the claims have not been supported by evidence at the molecular and functional levels. Here, we report on the bio-guided fractionation of H. noldeae and assessment of the inhibitory properties of some fractions and purified compounds on NLRP3 inflammasome and Interleukin 6 (IL-6). The activation of the NLRP3 inflammasome was determined by detecting the activity of caspase-1 and the production of Interleukin 1ß (IL-1ß) in Lipopolysaccharide (LPS) and ATP-stimulated Tamm-Horsfall Protein 1 (THP-1) macrophages, while the production of IL-6 was studied in LPS-stimulated RAW264.7 mouse macrophages. It was observed that hexane and ethyl acetate fractions of the crude extract of the aerial parts of H. noldeae, as well as caffeic acid, isoquercetin, and ER2.4 and ER2.7 fractions revealed significant inhibitory effects on Caspase-1 activities, and on IL-1ß and IL-6 production. The ER2.4 and ER2.7 fractions downregulated the production of IL-1ß and IL-6, in a similar range as the caspase-1 inhibitor AC-YVAD-CHO and the drug Dexamethasone, both used as controls, respectively. Overall, our work does provide the very first scientific based evidence for Hibiscus noldeae anti-inflammatory effects and widespread use by traditional healers in Rwanda for a variety of ailments.
Asunto(s)
Antiinflamatorios/farmacología , Hibiscus/química , Inflamasomas/efectos de los fármacos , Inflamación/tratamiento farmacológico , Interleucina-6/antagonistas & inhibidores , Proteína con Dominio Pirina 3 de la Familia NLR/antagonistas & inhibidores , Extractos Vegetales/farmacología , Animales , Inflamasomas/inmunología , Inflamasomas/metabolismo , Inflamación/inmunología , Inflamación/metabolismo , Inflamación/patología , Interleucina-6/metabolismo , Macrófagos/efectos de los fármacos , Macrófagos/inmunología , Macrófagos/metabolismo , Ratones , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismo , Células RAW 264.7RESUMEN
Protein carbamylation by cyanate is a post-translational modification associated with several (patho)physiological conditions, including cardiovascular disorders. However, the biochemical pathways leading to protein carbamylation are incompletely characterized. This work demonstrates that the heme protein myeloperoxidase (MPO), which is secreted at high concentrations at inflammatory sites from stimulated neutrophils and monocytes, is able to catalyze the two-electron oxidation of cyanide to cyanate and promote the carbamylation of taurine, lysine, and low-density lipoproteins. We probed the role of cyanide as both electron donor and low-spin ligand by pre-steady-state and steady-state kinetic analyses and analyzed reaction products by MS. Moreover, we present two further pathways of carbamylation that involve reaction products of MPO, namely oxidation of cyanide by hypochlorous acid and reaction of thiocyanate with chloramines. Finally, using an in vivo approach with mice on a high-fat diet and carrying the human MPO gene, we found that during chronic exposure to cyanide, mimicking exposure to pollution and smoking, MPO promotes protein-bound accumulation of carbamyllysine (homocitrulline) in atheroma plaque, demonstrating a link between cyanide exposure and atheroma. In summary, our findings indicate that cyanide is a substrate for MPO and suggest an additional pathway for in vivo cyanate formation and protein carbamylation that involves MPO either directly or via its reaction products hypochlorous acid or chloramines. They also suggest that chronic cyanide exposure could promote the accumulation of carbamylated proteins in atherosclerotic plaques.
Asunto(s)
Cianatos , Cianuros , Peroxidasa , Placa Aterosclerótica/enzimología , Carbamilación de Proteína , Animales , Citrulina/análogos & derivados , Citrulina/química , Citrulina/genética , Citrulina/metabolismo , Cianatos/química , Cianatos/metabolismo , Cianuros/química , Cianuros/metabolismo , Humanos , Ratones , Ratones Noqueados , Oxidación-Reducción , Peroxidasa/química , Peroxidasa/genética , Peroxidasa/metabolismo , Placa Aterosclerótica/genética , Placa Aterosclerótica/patologíaRESUMEN
Interleukin-21 (IL-21) is a cytokine with potent regulatory effects on different immune cells. Recently, IL-21 has been contemplated for use in the treatment of cancers. However, the molecular mechanisms regulating human IL-21 gene expression has not yet been described. In this study, we initially studied the promoter region and identified the transcription start site. We thereafter described the essential region upstream of the transcription start site and showed the in vivo binding of NFATc2 and SP1 transcription factors to this region, in addition to their positive role in IL-21 expression. We also studied the role of microRNAs (miRNAs) in regulating IL-21 expression. We, thus, established the miRNA profile of CD4+CD45RO+ versus CD4+CD45RA+ isolated from healthy volunteers and identified a signature composed of 12 differentially expressed miRNAs. We showed that miR-302c is able to negatively regulate IL-21 expression by binding directly to its target site in the 3'-untranslated region. Moreover, after using fresh human CD4-positive T cells, we observed the high acetylation level of histone H4, an observation well in line with the already described high expression of IL-21 in CD4+CD45RO+ versus CD4+CD45RA+ T cells. Altogether, our data identified different molecular mechanisms regulating IL-21 expression.
Asunto(s)
Linfocitos T CD4-Positivos/metabolismo , Interleucinas/metabolismo , Antígenos Comunes de Leucocito/metabolismo , MicroARNs/metabolismo , Factores de Transcripción NFATC/metabolismo , Factor de Transcripción Sp1/metabolismo , Regiones no Traducidas 3' , Acetilación , Sitios de Unión , Linfocitos T CD4-Positivos/inmunología , Células HEK293 , Células HeLa , Voluntarios Sanos , Histonas/metabolismo , Humanos , Interleucinas/genética , Células Jurkat , Antígenos Comunes de Leucocito/inmunología , MicroARNs/genética , Factores de Transcripción NFATC/genética , Regiones Promotoras Genéticas , Factor de Transcripción Sp1/genética , Sitio de Iniciación de la Transcripción , Transcripción GenéticaRESUMEN
Myeloperoxidase (MPO) is a member of the mammalian peroxidase family. It is mainly expressed in neutrophils, monocytes and macrophages. As a catalyzer of reactive oxidative species and radical species formation, it contributes to neutrophil bactericidal activity. Nevertheless MPO invalidation does not seem to have major health consequences in affected individuals. This suggests that MPO might have alternative functions supporting its conservation during evolution. We will review the available data supporting these non-canonical functions in terms of tissue specific expression, function and enzymatic activity. Thus, we discuss its cell type specific expression. We review in between others its roles in angiogenesis, endothelial (dys-) function, immune reaction, and inflammation. We summarize its pathological actions in clinical conditions such as cardiovascular disease and cancer.
Asunto(s)
Enfermedades Cardiovasculares/inmunología , Inflamación/inmunología , Neoplasias/inmunología , Peroxidasa/inmunología , Inmunidad Adaptativa , Animales , Enfermedades Cardiovasculares/enzimología , Enfermedades Cardiovasculares/metabolismo , Enfermedades Cardiovasculares/patología , Células Endoteliales/enzimología , Células Endoteliales/inmunología , Células Endoteliales/metabolismo , Células Endoteliales/patología , Humanos , Inmunidad Innata , Inflamación/enzimología , Inflamación/metabolismo , Inflamación/patología , Neoplasias/enzimología , Neoplasias/metabolismo , Neoplasias/patología , Neovascularización Fisiológica , Neutrófilos/enzimología , Neutrófilos/inmunología , Neutrófilos/metabolismo , Neutrófilos/patología , Peroxidasa/análisis , Peroxidasa/metabolismoRESUMEN
Myeloperoxidase is a member of the mammalian peroxidase family, mainly expressed in the myeloblastic cell lineage. It is considered a major bactericidal agent as it is released in the phagosome where it catalyzes the formation of reactive oxygen species. It is also released in the extracellular spaces including blood where it is absorbed on (lipo)proteins and endothelial cell surface, interfering with endothelial function. We performed RNA sequencing on MPO-treated endothelial cells, analyzed their transcriptome and validated the profile of gene expression by individual qRT-PCR. Some of the induced genes could be grouped in several functional networks, including tubulogenesis, angiogenesis, and blood vessel morphogenesis and development as well as signal transduction pathways associated to these mechanisms. MPO treatment mimicked the effects of VEGF on several signal transduction pathways, such as Akt, ERK or FAK involved in angiogenesis. Accordingly MPO, independently of its enzymatic activity, stimulated tube formation by endothelial cells. RNA interference also pointed at a role of endogenous MPO in tubulogenesis and endothelium wound repair in vitro. These data suggest that MPO, whether from endogenous or exogenous sources, could play a role in angiogenesis and vascular repair in vivo.
Asunto(s)
Endotelio Vascular/enzimología , Sistema de Señalización de MAP Quinasas , Peroxidasa/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , Línea Celular Transformada , Humanos , Neovascularización Patológica/metabolismo , Procesamiento Proteico-Postraduccional , TranscriptomaRESUMEN
Myeloperoxidase (MPO) is able to promote several kinds of damage and is involved in mechanisms leading to various diseases such as atherosclerosis or cancers. An example of these damages is the chlorination of nucleic acids, which is considered as a specific marker of the MPO activity. Since 5-chlorocytidine has been recently shown in healthy donor plasmas, this study aimed at discovering if these circulating modified nucleosides could be incorporated into RNA and DNA and if their presence impacts the ability of enzymes involved in the incorporation, transcription, and translation processes. Experimentations, which were carried out in vitro with endothelial and prostatic cells, showed a large penetration of all chloronucleosides but an exclusive incorporation of 5-chlorocytidine into RNA. However, no incorporation into DNA was observed. This specific incorporation is accompanied by an important reduction of translation yield. Although, in vitro, DNA polymerase processed in the presence of chloronucleosides but more slowly than in control conditions, ribonucleotide reductase could not reduce chloronucleotides prior to the replication. This reduction seems to be a limiting step, protecting DNA from chloronucleoside incorporation. This study shows the capacity of transcription enzyme to specifically incorporate 5-chlorocytidine into RNA and the loss of capacity-complete or partial-of different enzymes, involved in replication, transcription or translation, in the presence of chloronucleosides. Questions remain about the long-term impact of such specific incorporation in the RNA and such decrease of protein production on the cell viability and function.
Asunto(s)
Células Endoteliales/citología , Líquido Extracelular/química , Nucleósidos/química , Próstata/citología , ARN/análisis , Células Cultivadas , Cloro/química , Citidina/química , Halogenación , Humanos , Masculino , Nucleósidos/sangre , Peroxidasa/metabolismo , Biosíntesis de Proteínas , ARN/química , Transcripción GenéticaRESUMEN
African trypanosomes survive the immune defense of their hosts by regularly changing their antigenic coat made of variant surface glycoprotein (VSG). The Trypanosoma brucei genome contains more than 1,000 VSG genes. To be expressed, a given VSG gene must be located in one of 15 telomeric regions termed "VSG expression sites" (ESs), each of which contains a polycistronic transcription unit that includes ES-associated genes. Only one ES is fully active at a time, so only one VSG gene is transcribed per cell. Although this monoallelic expression is controlled at the transcriptional level, the precise molecular mechanism for this control is not understood. Here we report that in single cells transcription is initiated on several ESs simultaneously, indicating that the monoallelic control is not determined only at transcription initiation, but also at further control steps such as transcription elongation or RNA processing.
Asunto(s)
Transcripción Genética , Trypanosoma brucei brucei/metabolismo , Glicoproteínas Variantes de Superficie de Trypanosoma/genética , Alelos , Variación Antigénica , Línea Celular , Cartilla de ADN , Genes Protozoarios , Variación Genética , Humanos , Reacción en Cadena de la Polimerasa , Tripanosomiasis Africana/sangre , Tripanosomiasis Africana/parasitología , Glicoproteínas Variantes de Superficie de Trypanosoma/metabolismoRESUMEN
BACKGROUND: In order to promote infection, the blood-borne parasite Trypanosoma brucei releases factors that upregulate arginase expression and activity in myeloid cells. METHODOLOGY/PRINCIPAL FINDINGS: By screening a cDNA library of T. brucei with an antibody neutralizing the arginase-inducing activity of parasite released factors, we identified a Kinesin Heavy Chain isoform, termed TbKHC1, as responsible for this effect. Following interaction with mouse myeloid cells, natural or recombinant TbKHC1 triggered SIGN-R1 receptor-dependent induction of IL-10 production, resulting in arginase-1 activation concomitant with reduction of nitric oxide (NO) synthase activity. This TbKHC1 activity was IL-4Rα-independent and did not mirror M2 activation of myeloid cells. As compared to wild-type T. brucei, infection by TbKHC1 KO parasites was characterized by strongly reduced parasitaemia and prolonged host survival time. By treating infected mice with ornithine or with NO synthase inhibitor, we observed that during the first wave of parasitaemia the parasite growth-promoting effect of TbKHC1-mediated arginase activation resulted more from increased polyamine production than from reduction of NO synthesis. In late stage infection, TbKHC1-mediated reduction of NO synthesis appeared to contribute to liver damage linked to shortening of host survival time. CONCLUSION: A kinesin heavy chain released by T. brucei induces IL-10 and arginase-1 through SIGN-R1 signaling in myeloid cells, which promotes early trypanosome growth and favors parasite settlement in the host. Moreover, in the late stage of infection, the inhibition of NO synthesis by TbKHC1 contributes to liver pathogenicity.
Asunto(s)
Arginasa/inmunología , Cinesinas/inmunología , Proteínas Protozoarias/inmunología , Trypanosoma brucei brucei/inmunología , Tripanosomiasis Africana/inmunología , Animales , Arginasa/genética , Moléculas de Adhesión Celular/genética , Moléculas de Adhesión Celular/inmunología , Activación Enzimática/genética , Activación Enzimática/inmunología , Interleucina-10/genética , Interleucina-10/inmunología , Cinesinas/genética , Lectinas Tipo C/genética , Lectinas Tipo C/inmunología , Ratones , Ratones Noqueados , Óxido Nítrico/genética , Óxido Nítrico/inmunología , Proteínas Protozoarias/genética , Receptores de Superficie Celular/genética , Receptores de Superficie Celular/inmunología , Trypanosoma brucei brucei/genética , Tripanosomiasis Africana/genética , Tripanosomiasis Africana/patologíaRESUMEN
BACKGROUND: It is a known fact that blood flow pattern and more specifically the pulsatile time variation of shear stress on the vascular wall play a key role in atherogenesis. The paper presents the conception, the building and the control of a new in vitro test bench that mimics the pulsatile flows behavior based on in vivo measurements. METHODS: An in vitro cardiovascular simulator is alimented with in vivo constraints upstream and provided with further post-processing analysis downstream in order to mimic the pulsatile in vivo blood flow quantities. This real-time controlled system is designed to perform real pulsatile in vivo blood flow signals to study endothelial cells' behavior under near physiological environment. The system is based on an internal model controller and a proportional-integral controller that controls a linear motor with customized piston pump, two proportional-integral controllers that control the mean flow rate and temperature of the medium. This configuration enables to mimic any resulting blood flow rate patterns between 40 and 700 ml/min. In order to feed the system with reliable periodic flow quantities in vivo measurements were performed. Data from five patients (1 female, 4 males; ages 44-63) were filtered and post-processed using the Newtonian Womersley's solution. These resulting flow signals were compared with 2D axisymmetric, numerical simulation using a Carreau non-Newtonian model to validate the approximation of a Newtonian behavior. RESULTS: This in vitro test bench reproduces the measured flow rate time evolution and the complexity of in vivo hemodynamic signals within the accuracy of the relative error below 5%. CONCLUSIONS: This post-processing method is compatible with any real complex in vivo signal and demonstrates the heterogeneity of pulsatile patterns in coronary arteries among of different patients. The comparison between analytical and numerical solution demonstrate the fair quality of the Newtonian Womersley's approximation. Therefore, Womersley's solution was used to calculate input flow rate for the in vitro test bench.
Asunto(s)
Vasos Coronarios/fisiología , Procesamiento de Señales Asistido por Computador , Adulto , Velocidad del Flujo Sanguíneo , Vasos Coronarios/diagnóstico por imagen , Femenino , Análisis de Fourier , Humanos , Hidrodinámica , Masculino , Persona de Mediana Edad , Modelos Biológicos , Flujo Pulsátil , Reproducibilidad de los Resultados , Tomografía Computarizada por Rayos XRESUMEN
Oxidation of LDL by the myeloperoxidase (MPO)-H2O2-chloride system is a key event in the development of atherosclerosis. The present study aimed at investigating the interaction of MPO with native and modified LDL and at revealing posttranslational modifications on apoB-100 (the unique apolipoprotein of LDL) in vitro and in vivo. Using amperometry, we demonstrate that MPO activity increases up to 90% when it is adsorbed at the surface of LDL. This phenomenon is apparently reflected by local structural changes in MPO observed by circular dichroism. Using MS, we further analyzed in vitro modifications of apoB-100 by hypochlorous acid (HOCl) generated by the MPO-H2O2-chloride system or added as a reagent. A total of 97 peptides containing modified residues could be identified. Furthermore, differences were observed between LDL oxidized by reagent HOCl or HOCl generated by the MPO-H2O2-chloride system. Finally, LDL was isolated from patients with high cardiovascular risk to confirm that our in vitro findings are also relevant in vivo. We show that several HOCl-mediated modifications of apoB-100 identified in vitro were also present on LDL isolated from patients who have increased levels of plasma MPO and MPO-modified LDL. In conclusion, these data emphasize the specificity of MPO to oxidize LDL.
Asunto(s)
Apolipoproteína B-100/metabolismo , Lipoproteínas LDL/metabolismo , Peroxidasa/metabolismo , Secuencia de Aminoácidos , Apolipoproteína B-100/química , Estudios de Casos y Controles , Humanos , Peróxido de Hidrógeno/química , Hidrólisis , Enfermedades Renales/sangre , Enfermedades Renales/terapia , Lipoproteínas LDL/química , Datos de Secuencia Molecular , Oxidación-Reducción , Fragmentos de Péptidos , Peroxidasa/química , Procesamiento Proteico-Postraduccional , Diálisis RenalRESUMEN
FKBP12 proteins are able to inhibit TOR kinases or calcineurin phosphatases upon binding of rapamycin or FK506 drugs, respectively. The Trypanosoma brucei FKBP12 homologue (TbFKBP12) was found to be a cytoskeleton-associated protein with specific localization in the flagellar pocket area of the bloodstream form. In the insect procyclic form, RNA interference-mediated knockdown of TbFKBP12 affected motility. In bloodstream cells, depletion of TbFKBP12 affected cytokinesis and cytoskeleton architecture. These last effects were associated with the presence of internal translucent cavities limited by an inside-out configuration of the normal cell surface, with a luminal variant surface glycoprotein coat lined up by microtubules. These cavities, which recreated the streamlined shape of the normal trypanosome cytoskeleton, might represent unsuccessful attempts for cell abscission. We propose that TbFKBP12 differentially affects stage-specific processes through association with the cytoskeleton.
Asunto(s)
Citocinesis , Proteínas Protozoarias/fisiología , Proteína 1A de Unión a Tacrolimus/metabolismo , Trypanosoma brucei brucei/enzimología , Movimiento Celular , ADN de Cinetoplasto/metabolismo , ADN Protozoario/metabolismo , Flagelos/metabolismo , Flagelos/ultraestructura , Técnicas de Silenciamiento del Gen , Microtúbulos/metabolismo , Transporte de Proteínas , Proteínas Protozoarias/genética , ARN Interferente Pequeño/genética , Proteína 1A de Unión a Tacrolimus/genética , Trypanosoma brucei brucei/crecimiento & desarrollo , Trypanosoma brucei brucei/ultraestructuraRESUMEN
Cardiovascular disease linked to atherosclerosis is the leading cause of death worldwide. Atherosclerosis is mainly linked to dysfunction in vascular endothelial cells and subendothelial accumulation of oxidized forms of LDL. In the present study, we investigated the role of myeloperoxidase oxidized LDL (Mox-LDL) in endothelial cell dysfunction. We studied the effect of proinflammatory Mox-LDL treatment on endothelial cell motility, a parameter essential for normal vascular processes such as angiogenesis and blood vessel repair. This is particularly important in the context of an atheroma plaque, where vascular wall integrity is affected and interference with its repair could contribute to progression of the disease. We investigated in vitro the effect of Mox-LDL on endothelial cells angiogenic properties and we also studied the signalling pathways that could be affected by analysing Mox-LDL effect on the expression of angiogenesis-related genes. We report that Mox-LDL inhibits endothelial cell motility and tubulogenesis through an increase in miR-22 and heme oxygenase 1 expression. Our in vitro data indicate that Mox-LDL interferes with parameters associated with angiogenesis. They suggest that high LDL levels in patients would impair their endothelial cell capacity to cope with a damaged endothelium contributing negatively to the progression of the atheroma plaque.
Asunto(s)
Células Endoteliales/citología , Endotelio Vascular/metabolismo , Hemo-Oxigenasa 1/metabolismo , Lipoproteínas LDL/metabolismo , MicroARNs/metabolismo , Peroxidasa/metabolismo , Animales , Células CHO , Movimiento Celular , Cricetinae , Cricetulus , Progresión de la Enfermedad , Células Endoteliales de la Vena Umbilical Humana/citología , Humanos , Neovascularización Patológica , Placa Aterosclerótica/metabolismo , Transducción de Señal , Lesiones del Sistema Vascular/metabolismo , Cicatrización de HeridasRESUMEN
Prdm12 is an epigenetic regulator expressed in developing and mature nociceptive neurons, playing a key role in their specification during neurogenesis and modulating pain sensation at adulthood. In vitro studies suggested that Prdm12 recruits the methyltransferase G9a through its zinc finger domains to regulate target gene expression, but how Prdm12 interacts with G9a and whether G9a plays a role in Prdm12's functional properties in sensory ganglia remain unknown. Here we report that Prdm12-G9a interaction is likely direct and that it involves the SET domain of G9a. We show that both proteins are largely co-expressed in dorsal root ganglia during early murine development, opening the possibility that G9a plays a role in DRG and may act as a mediator of Prdm12's function in the development of nociceptive sensory neurons. To test this hypothesis, we conditionally inactivated G9a in neural crest using a Wnt1-Cre transgenic mouse line. We found that the specific loss of G9a in the neural crest lineage does not lead to dorsal root ganglia hypoplasia due to the loss of somatic nociceptive neurons nor to the ectopic expression of the visceral determinant Phox2b as observed upon Prdm12 ablation. These findings suggest that Prdm12 function in the initiation of the nociceptive lineage does not critically involves its interaction with G9a.
Asunto(s)
Neurogénesis , Nociceptores , Ratones , Animales , Nociceptores/metabolismo , Neurogénesis/fisiología , Células Receptoras Sensoriales , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Ganglios Espinales , Ratones Transgénicos , Proteínas Portadoras/genética , Proteínas Portadoras/metabolismo , Proteínas del Tejido Nervioso/metabolismoRESUMEN
BACKGROUND: MicroRNAs (miRNAs) are small (19-22-nt) single-stranded noncoding RNA molecules whose deregulation of expression can contribute to human disease including the multistep processes of carcinogenesis in human. Circulating miRNAs are emerging biomarkers in many diseases and cancers such as type 2 diabetes, pulmonary disease, colorectal cancer, and gastric cancer among others; however, defining a plasma miRNA signature in acute myeloblastic leukemia (AML) that could serve as a biomarker for diagnosis or in the follow-up has not been done yet. METHODS: TaqMan miRNA microarray was performed to identify deregulated miRNAs in the plasma of AML patients. Quantitative real-time RT-PCR was used to validate the results. Receiver-operator characteristic (ROC) curve analysis was conducted to evaluate the diagnostic accuracy of the highly and significantly identified deregulated miRNA(s) as potential candidate biomarker(s). RESULTS: The plasma expression level of let-7d, miR-150, miR-339, and miR-342 was down-regulated whilst that of let-7b, and miR-523 was up-regulated in the AML group at diagnosis compared to healthy controls. ROC curve analyses revealed an AUC (the areas under the ROC curve) of 0.835 (95% CI: 0.7119- 0.9581; P<0.0001) and 0.8125 (95% CI: 0.6796-0.9454; P=0.0005) for miR-150, and miR-342 respectively. Combined ROC analyses using these 2 miRNAs revealed an elevated AUC of 0.86 (95% CI: 0.7819-0.94; P<0.0001) indicating the additive effect in the diagnostic value of these 2 miRNAs. QRT-PCR results showed that the expression level of these two miRs in complete remission AML patients resembled that of healthy controls. CONCLUSIONS: Our findings indicated that plasma miR-150 and miR-342 are novel important promising biomarkers in the diagnosis of AML. These novel and promising markers warrant validation in larger prospective studies.
Asunto(s)
Biomarcadores de Tumor/sangre , Leucemia Mieloide Aguda/sangre , MicroARNs/sangre , Área Bajo la Curva , Biomarcadores de Tumor/genética , Estudios de Casos y Controles , Perfilación de la Expresión Génica , Humanos , Leucemia Mieloide Aguda/genética , MicroARNs/genética , Análisis de Secuencia por Matrices de Oligonucleótidos , Curva ROC , Reacción en Cadena en Tiempo Real de la PolimerasaRESUMEN
Oxidation of low-density lipoprotein (LDL) has a key role in atherogenesis. Among the different models of oxidation that have been studied, the one using myeloperoxidase (MPO) is thought to be more physiopathologically relevant. Apolipoprotein B-100 is the unique protein of LDL and is the major target of MPO. Furthermore, MPO rapidly adsorbs at the surface of LDL, promoting oxidation of amino acid residues and formation of oxidized lipoproteins that are commonly named Mox-LDL. The latter is not recognized by the LDL receptor and is accumulated by macrophages. In the context of atherogenesis, Mox-LDL accumulates in macrophages leading to foam cell formation. Furthermore, Mox-LDL seems to have specific effects and triggers inflammation. Indeed, those oxidized lipoproteins activate endothelial cells and monocytes/macrophages and induce proinflammatory molecules such as TNF α and IL-8. Mox-LDL may also inhibit fibrinolysis mediated via endothelial cells and consecutively increase the risk of thrombus formation. Finally, Mox-LDL has been involved in the physiopathology of several diseases linked to atherosclerosis such as kidney failure and consequent hemodialysis therapy, erectile dysfunction, and sleep restriction. All these issues show that the investigations of MPO-dependent LDL oxidation are of importance to better understand the inflammatory context of atherosclerosis.
Asunto(s)
Inflamación/metabolismo , Lipoproteínas LDL/metabolismo , Peroxidasa/metabolismo , Apolipoproteína B-100/metabolismo , Aterosclerosis , Endocitosis , Disfunción Eréctil/metabolismo , Hígado Graso/metabolismo , Femenino , Fibrinólisis , Humanos , Peróxido de Hidrógeno/química , Interleucina-8/metabolismo , Macrófagos/metabolismo , Masculino , Oxígeno/química , Enfermedad Pulmonar Obstructiva Crónica/metabolismo , Diálisis Renal , Trastornos del Sueño-Vigilia/metabolismo , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
The present paradigm of atherogenesis proposes that low density lipoproteins (LDLs) are trapped in subendothelial space of the vascular wall where they are oxidized. Previously, we showed that oxidation is not restricted to the subendothelial location. Myeloperoxidase (MPO), an enzyme secreted by neutrophils and macrophages, can modify LDL (Mox-LDL) at the surface of endothelial cells. In addition we observed that the activation of the endothelial cells by angiotensin II amplifies this process. We suggested that induction of the NADPH oxidase complex was a major step in the oxidative process. Based on these data, we asked whether there was an independent association, in 121 patients, between NADPH oxidase modulators, such as angiotensin II, adiponectin, and levels of circulating Mox-LDL. Our observations suggest that the combination of blood angiotensin II, MPO activity, and adiponectin explains, at least partially, serum Mox-LDL levels.
Asunto(s)
Adiponectina/sangre , Angiotensina II/sangre , Lipoproteínas LDL/sangre , Peroxidasa/metabolismo , Adulto , Anciano , Apolipoproteínas B/sangre , Estudios Transversales , Humanos , Síntomas del Sistema Urinario Inferior/sangre , Masculino , Persona de Mediana Edad , NADPH Oxidasas/metabolismo , Peroxidasas/metabolismoRESUMEN
Onchocerciasis is an infectious disease of public health and socio-economic importance in most parts of Sub-Saharan Africa. The objective of this study was to evaluate the effects of the suspension of implementation activities towards combating onchocerciasis in the Bandjoun and Massangam health districts in the West Region of Cameroon as a consequence of the COVID-19 pandemic. Data on socio-demographic and clinical characteristics were obtained using a structured questionnaire. All participants in both health districts were examined for the presence of clinical manifestations of onchocerciasis. In addition, two skin snips were obtained from the knee of each participant and examined for the presence of microfilaria. All data were categorized, coded, entered in a database, and analysed using SPSS version 23.0. A total of 229 participants in the Bandjoun health district and 378 in the Massangam health district were recruited for the study. In both health districts, there was no significant difference between male and female participants in terms of the clinical manifestations of onchocerciasis. The prevalence of nodules was 8.7% in the Bandjoun health district and 20.6% in the Massangam health district while the prevalence of microfilaria carriers in Bandjoun and Massangam health districts was 3.5% and 3.7%, respectively. Except for the Tsesse and Lemgo communities in the Bandjoun health district, there was a reduction in the prevalence of microfilaria in the communities that were studied when compared to previous data obtained before the disruption of control programmes activities. Overall, in both health districts, elderly individuals bear the largest burden of onchocerciasis. Based on the results obtained, we conclude that the temporary suspension of Neglected Tropical Disease control programme activities by the World Head Organization as a result of COVID-19 may have resulted to recrudescence of O. volvulus transmission in hypoendemic communities in the Bandjoun health district.
Asunto(s)
COVID-19 , Oncocercosis , Animales , Humanos , Masculino , Femenino , Anciano , Oncocercosis/tratamiento farmacológico , Oncocercosis/epidemiología , Oncocercosis/prevención & control , Ivermectina/uso terapéutico , Administración Masiva de Medicamentos , Camerún/epidemiología , Prevalencia , Pandemias , COVID-19/epidemiología , MicrofilariasRESUMEN
Myeloperoxidase (MPO) has been reported in prostate tissue, and considering its pro-oxidant properties, this location might be linked to prostate pathology. The possibility that the glandular prostatic tissue might be the source of MPO and its potential inflammatory effects must be tested. Human prostate material was obtained from prostate biopsies and radical prostatectomies. Immunohistochemistry was performed using MPO-specific human antibody. In situ hybridization using MPO-specific probes and laser-assisted microdissection for quantitative real-time RT-PCR were performed to observe whether MPO is being produced in prostate tissue. Mass spectrometry on prostate biopsies was used to detect products of MPO activity in nucleic acids (DNA/RNA). MPO contribution to intracellular accumulation of ROS and interleukin-8 in prostatic epithelial cells was monitored in vitro. Immunohistochemistry confirmed cellular localization of MPO in epithelial cells of the prostate. The staining varied from light to high intensity. In situ hybridization did not address the presence of mRNA coding for MPO. No MPO-specific modifications on nucleic acids were detected. Mox-LDL was a major factor inducing ROS and cytokines production in prostatic epithelial cells. We did not demonstrate that MPO was synthetized by prostatic epithelial cells. However, in vitro experiments showed the ability of MPO to potentiate the ROS production and inflammation on prostate epithelial cells. Results do not allow us to demonstrate a role of MPO in prostate to date but further studies are mandatory to focus on the potential impact of MPO in the development of prostatic diseases.
Asunto(s)
Peroxidasa , Próstata , Masculino , Humanos , Próstata/patología , Especies Reactivas de Oxígeno , Peroxidasa/análisis , Células Epiteliales/patología , ARN Mensajero/análisisRESUMEN
Onchocerciasis is a Neglected Tropical Disease that has a significant socioeconomic impact, especially in Sub-Saharan Africa. Numerous reports indicate that the Expanded Special Project for the Elimination of Neglected Tropical Diseases needs novel diagnostic tools before achieving its goal of successful elimination of onchocerciasis in Africa. The current diagnostic tests are either invasive, insensitive, or not applicable in the field and about 25% of persons infected cannot mount immune responses against the single antigen used in the only approved Ov-16 serological test. In the quest to identify novel biomarkers that can be used to certify that a patient is free from the disease, evaluate the progress of elimination programmes, and conduct post elimination surveillances, mass spectrometric analysis of Onchocerca volvulus crude extract revealed that 1392 proteins are expressed in the adult and microfilariae stages of the parasite. Computational analysis predicted six of the proteins as O. volvulus potential diagnostic targets. Linear B-epitopes were predicted from the six proteins and used to construct a multiepitope antigen (OvMCBL02). Serological analysis revealed that the OvMCBL02 test significantly differentiated between serum samples of onchocerciasis patients from the Kombone Health Area in the South West Region of Cameroon (n = 63) and control serum samples from Rwanda (n = 29) and Europe (n = 26) as well as between serum samples from the onchocerciasis hyperendemic region of Kombone Health Area (n = 63) and the hypoendemic region of Bandjoun Health District (n = 54). Interestingly, the test did not cross-react with serum samples from patients suffering from related nematode infections, thereby suggesting that further characterization of the OvMCBL02 multiepitope antigen will render it an additional member of the diagnostic toolbox for the elimination of onchocerciasis.