RESUMEN
AIMS: According to Braak's hypothesis, it is plausible that Parkinson's disease (PD) originates in the enteric nervous system (ENS) and spreads to the brain through the vagus nerve. In this work, we studied whether inflammatory bowel diseases (IBDs) in humans can progress with the emergence of pathogenic α-synuclein (α-syn) in the gastrointestinal tract and midbrain dopaminergic neurons. METHODS: We have analysed the gut and the ventral midbrain from subjects previously diagnosed with IBD and form a DSS-based rat model of gut inflammation in terms of α-syn pathology. RESULTS: Our data support the existence of pathogenic α-syn in both the gut and the brain, thus reinforcing the potential role of the ENS as a contributing factor in PD aetiology. Additionally, we have analysed the effect of a DSS-based rat model of gut inflammation to demonstrate (i) the appearance of P-α-syn inclusions in both Auerbach's and Meissner's plexuses (gut), (ii) an increase in α-syn expression in the ventral mesencephalon (brain) and (iii) the degeneration of nigral dopaminergic neurons, which all are considered classical hallmarks in PD. CONCLUSION: These results strongly support the plausibility of Braak's hypothesis and emphasise the significance of peripheral inflammation and the gut-brain axis in initiating α-syn aggregation and transport to the substantia nigra, resulting in neurodegeneration.
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Enfermedades Inflamatorias del Intestino , Enfermedad de Parkinson , Humanos , Ratas , Animales , alfa-Sinucleína/metabolismo , Enfermedad de Parkinson/patología , Encéfalo/patología , Inflamación/patología , Neuronas Dopaminérgicas/metabolismo , Enfermedades Inflamatorias del Intestino/patologíaRESUMEN
Water is an abundant and important component of the human brain, the homeostasis of which is rigorously controlled [...].
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Acuaporinas , Encefalopatías , Humanos , Acuaporinas/metabolismo , Agua/metabolismo , Homeostasis , Encéfalo/metabolismoRESUMEN
HIV-1 virion production is driven by Gag and Gag-Pol (GP) proteins, with Gag forming the bulk of the capsid and driving budding, while GP binds Gag to deliver the essential virion enzymes protease, reverse transcriptase, and integrase. Virion GP levels are traditionally thought to reflect the relative abundances of GP and Gag in cells (â¼1:20), dictated by the frequency of a -1 programmed ribosomal frameshifting (PRF) event occurring in gag-pol mRNAs. Here, we exploited a panel of PRF mutant viruses to show that mechanisms in addition to PRF regulate GP incorporation into virions. First, we show that GP is enriched â¼3-fold in virions relative to cells, with viral infectivity being better maintained at subphysiological levels of GP than when GP levels are too high. Second, we report that GP is more efficiently incorporated into virions when Gag and GP are synthesized in cis (i.e., from the same gag-pol mRNA) than in trans, suggesting that Gag/GP translation and assembly are spatially coupled processes. Third, we show that, surprisingly, virions exhibit a strong upper limit to trans-delivered GP incorporation; an adaptation that appears to allow the virus to temper defects to GP/Gag cleavage that may negatively impact reverse transcription. Taking these results together, we propose a "weighted Goldilocks" scenario for HIV-1 GP incorporation, wherein combined mechanisms of GP enrichment and exclusion buffer virion infectivity over a broad range of local GP concentrations. These results provide new insights into the HIV-1 virion assembly pathway relevant to the anticipated efficacy of PRF-targeted antiviral strategies. IMPORTANCE HIV-1 infectivity requires incorporation of the Gag-Pol (GP) precursor polyprotein into virions during the process of virus particle assembly. Mechanisms dictating GP incorporation into assembling virions are poorly defined, with GP levels in virions traditionally thought to solely reflect relative levels of Gag and GP expressed in cells, dictated by the frequency of a -1 programmed ribosomal frameshifting (PRF) event that occurs in gag-pol mRNAs. Herein, we provide experimental support for a "weighted Goldilocks" scenario for GP incorporation, wherein the virus exploits both random and nonrandom mechanisms to buffer infectivity over a wide range of GP expression levels. These mechanistic data are relevant to ongoing efforts to develop antiviral strategies targeting PRF frequency and/or HIV-1 virion maturation.
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Sistema de Lectura Ribosómico , Regulación Viral de la Expresión Génica , Infecciones por VIH/virología , VIH-1/fisiología , Ensamble de Virus , Productos del Gen gag del Virus de la Inmunodeficiencia Humana/genética , Productos del Gen gag del Virus de la Inmunodeficiencia Humana/metabolismo , Inhibidores de la Proteasa del VIH/farmacología , VIH-1/efectos de los fármacos , Humanos , Secuencias Invertidas Repetidas , Modelos Biológicos , Conformación de Ácido Nucleico , Estabilidad del ARN , ARN Viral/química , ARN Viral/genética , Virión , Replicación ViralRESUMEN
Metabolites produced by an altered gut microbiota might mediate the effects in the brain. Among metabolites, the fecal volatile organic compounds (VOCs) are considered to be potential biomarkers. In this study, we examined both the VOCs and bacterial taxa in the feces from healthy subjects and Alzheimer's disease (AD) patients at early and middle stages. Remarkably, 29 fecal VOCs and 13 bacterial genera were differentiated from the healthy subjects and the AD patients. In general, higher amounts of acids and esters were found in in the feces of the AD patients and terpenes, sulfur compounds and aldehydes in the healthy subjects. At the early stage of AD, the most relevant VOCs with a higher abundance were short-chain fatty acids and their producing bacteria, Faecalibacterium and Lachnoclostridium. Coinciding with the development of dementia in the AD patients, parallel rises of heptanoic acid and Peptococcus were observed. At a more advanced stage of AD, the microbiota and volatiles shifted towards a profile in the feces with increases in hexanoic acid, Ruminococcus and Blautia. The most remarkable VOCs that were associated with the healthy subjects were 4-ethyl-phenol and dodecanol, together with their possible producers Clostridium and Coprococcus. Our results revealed a VOCs and microbiota crosstalk in AD development and their profiles in the feces were specific depending on the stage of AD. Additionally, some of the most significant fecal VOCs identified in our study could be used as potential biomarkers for the initiation and progression of AD.
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Enfermedad de Alzheimer , Disfunción Cognitiva , Microbiota , Compuestos Orgánicos Volátiles , Humanos , Compuestos Orgánicos Volátiles/metabolismo , Enfermedad de Alzheimer/metabolismo , Disfunción Cognitiva/microbiología , Heces/microbiología , Ácidos Grasos Volátiles/metabolismo , Bacterias/metabolismo , Biomarcadores/metabolismoRESUMEN
Neuroinflammation underlies neurodegenerative diseases. Herein, we test whether acute colon inflammation activates microglia and astrocytes, induces neuroinflammation, disturbs neuron intrinsic electrical properties in the primary motor cortex, and alters motor behaviors. We used a rat model of acute colon inflammation induced by dextran sulfate sodium. Inflammatory mediators and microglial activation were assessed in the primary motor cortex by PCR and immunofluorescence assays. Electrophysiological properties of the motor cortex neurons were determined by whole-cell patch-clamp recordings. Motor behaviors were examined using open-field and rotarod tests. We show that the primary motor cortex of rats with acute colon inflammation exhibited microglial and astrocyte activation and increased mRNA abundance of interleukin-6, tumor necrosis factor-alpha, and both inducible and neuronal nitric oxide synthases. These changes were accompanied by a reduction in resting membrane potential and rheobase and increased input resistance and action potential frequency, indicating motor neuron hyperexcitability. In addition, locomotion and motor coordination were impaired. In conclusion, acute colon inflammation induces motor cortex microglial and astrocyte activation and inflammation, which led to neurons' hyperexcitability and reduced motor coordination performance. The described disturbances resembled some of the early features found in amyotrophic lateral sclerosis patients and animal models, suggesting that colon inflammation might be a risk factor for developing this disease.
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Colitis , Corteza Motora , Animales , Colitis/inducido químicamente , Colitis/patología , Humanos , Inflamación/patología , Corteza Motora/patología , Neuronas Motoras/patología , Enfermedades Neuroinflamatorias , RatasRESUMEN
We reported that Disabled-2 (Dab2) is located at the apical membrane in suckling rat intestine. Here, we discovered that, in colon of suckling and adult mouse and of adult human, Dab2 is only at lateral crypt cell membrane and colocalized with E-cadherin. Dab2 depletion in Caco-2 cells led to E-cadherin internalization indicating that its membrane location requires Dab2. In mice, we found that 3 days of dextran sulfate sodium-induced colitis increased Dab2/E-cadherin colocalization, which was decreased as colitis progressed to 6 and 9 days. In agreement, Dab2/E-cadherin colocalization increased in human mild and severe ulcerative colitis and in polyps, being reduced in colon adenocarcinomas, which even showed epithelial Dab2 absence and E-cadherin delocalization. Epithelial Dab2 decrement preceded that of E-cadherin. We suggest that Dab2, by inhibiting E-cadherin internalization, stabilizes adherens junctions, and its absence from the epithelium may contribute to development of colon inflammation and cancer.
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Proteínas Adaptadoras Transductoras de Señales/genética , Adenocarcinoma/genética , Proteínas Reguladoras de la Apoptosis/genética , Cadherinas/genética , Neoplasias del Colon/genética , Proteínas Adaptadoras del Transporte Vesicular/genética , Adenocarcinoma/patología , Anciano , Animales , Células CACO-2 , Colitis/inducido químicamente , Colitis/genética , Colitis/patología , Colon/metabolismo , Colon/patología , Neoplasias del Colon/patología , Sulfato de Dextran/toxicidad , Células Epiteliales/metabolismo , Células Epiteliales/patología , Femenino , Humanos , Inflamación/genética , Inflamación/patología , Mucosa Intestinal/metabolismo , Mucosa Intestinal/patología , Masculino , Ratones , Persona de Mediana Edad , Pólipos/genética , Pólipos/patología , RatasRESUMEN
We define social media as an interactive online platform that allows users to communicate and exchange knowledge. Educational and medical profiles have slowly emerged on different social media platforms, helping to teach about and publicize diverse aspects of medicine. Radiology is one of the specialties that could potentially benefit the most from social media, as the radiologist tends to have little outside-the-hospital representation. Progressively, audiovisual content has been gaining ground on social networks: Facebook, Twitter, Instagram, Youtube, TikTok, etc. Instagram appears to be ideally suited for radiology given its image-based nature. In addition, Instagram can also be used as a tool to help radiologists share and discuss radiological images, improve communication with clinicians and patients, advertise themselves and their specialty, and humanize their profession. Nevertheless, legal matters and privacy issues should always be taken into account when using these tools. In this overview, we describe the development of social networks and communication tools in our own radiology department, focusing especially on our Instagram account, as it has had a wide impact on our hospital and radiology residents around the country. We will also provide a summary of the various social media platforms used for radiology education along with their pros and cons, including useful tips for safe and efficient use.
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Radiología , Medios de Comunicación Sociales , Hospitales , Humanos , RadiólogosRESUMEN
The objective was to evaluate the effect of the incorporation of calcium hydroxyapatite particles (HAp) in the diametral tensile strength of a conventional type II glass ionomer (GC Gold Label 2) and a resin-modified glass ionomer cement (GC Gold Label 2 LC R). Two experimental HAp (E1HAp or E2HAp) were synthesized and characterized using X-ray diffraction and Confocal Raman spectroscopy. Both HAp were added into the powder of a conventional or resin-modified glass ionomer cement at 5 or 10 wt.%. A commercial HAp (CHAp) was used as reference material. For each glass ionomer cement, a group without the incorporation of HAp was used as a control. A universal testing machine was used for the mechanical test. The results were analyzed through a two-way ANOVA test followed by a complementary Tukey test. For all analyzes, the level of significance was set at α = 0.05. The average particle size for E1Hap was 15 µm, E2HAp was 35 µm and for CHAp was 1 µm. For conventional GIC, the addition of 10% E1HAp and 5% CHAp significantly increased the diametral tensile strength values (p ≤ 0.005). On the other hand, for the resin-modified GIC, except for the 5% E2HAp group, all experimental groups significantly reduced the values of diametral tensile strength (p ≤ 0.007). The addition of HAp improved the mechanical properties only for the conventional glass ionomer cement.
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Durapatita , Cementos de Ionómero Vítreo , Ensayo de Materiales , Resistencia a la TracciónRESUMEN
BACKGROUND: Adequate fluid resuscitation is paramount in the management of acute pancreatitis (AP). The aim of this study is to assess benefits and harms of fluid therapy protocols in patients with AP. METHODS: MEDLINE, Embase, Science Citation Index and clinical trial registries were searched for randomised clinical trials published before May 2020, assessing types of fluids, routes and rates of administration. RESULTS: A total 15 trials (1073 participants) were included. Age ranged from 38 to 73 years; follow-up period ranged from 0.5 to 6 months. Ringer lactate (RL) showed a reduced number of severe adverse events (SAE) when compared to normal saline (NS) (OR 0.48; 95%CI 0.29-0.81, p = 0.006); additionally, NS showed reduced SAE (RR 0.38; 95%IC 0.27-0.54, p < 0.001) and organ failure (RR 0.30; 95%CI 0.21-0.44, p < 0.001) in comparison with hydroxyethyl starch (HES). High fluid rate fluid infusion showed increased mortality (OR 2.88; 95%CI 1.41-5.88, p = 0.004), increased number of SAE (RR 1.42; 95%CI 1.04-1.93, p = 0.030) and higher incidence of sepsis (RR 2.80; 95%CI 1.51-5.19, p = 0.001) compared to moderate fluid rate infusion. CONCLUSIONS: In patients with AP, RL should be preferred over NS and HES should not be recommended. Based on low-certainty evidence, moderate-rate fluid infusion should be preferred over high-rate infusion.
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Pancreatitis , Sepsis , Niño , Preescolar , Humanos , Enfermedad Aguda , Fluidoterapia , Pancreatitis/diagnóstico , Pancreatitis/terapia , Protocolos ClínicosRESUMEN
A 12-year-old girl presented with headache, nasal voice, and anosmia. Magnetic resonance imaging demonstrated a 11 × 9 × 8-cm tumor extending from the nasal cavity to the frontal lobes. Histological analysis was consistent with transitional meningioma WHO grade I. As far as we know, this is the first reported case of a giant olfactory groove meningioma invading the nasal cavity through the anterior cranial base. Complete surgical removal was accomplished with excellent result via a staged endoscopic and transcranial approach. A microvascularized vastus laterallis flap was used for reconstruction. Surgical options and technical details in the management of this lesion are reviewed.
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Neoplasias Meníngeas , Meningioma , Neoplasias de la Base del Cráneo , Niño , Endoscopía , Femenino , Humanos , Neoplasias Meníngeas/diagnóstico por imagen , Neoplasias Meníngeas/cirugía , Meningioma/diagnóstico por imagen , Meningioma/cirugía , Cavidad Nasal/diagnóstico por imagen , Cavidad Nasal/cirugía , Base del Cráneo , Colgajos QuirúrgicosRESUMEN
BACKGROUND: Closure of the skin defect in myelomeningocele repair determines the quality of the surgical result. In large myelomeningoceles, the direct closure or by skin undermining may not be possible. In these cases, the skin defect must be closed using flaps to provide a tension-free and successful closure. METHODS: In this procedure, after neurosurgical repair and closure of the placode, the defect surgically becomes an ovoid in shape. Then, this defect is closed by transposition of two dorsal intercostal artery perforator propeller flaps. The defect size was > 5 cm in diameter. The localization of the lesion was lumbosacral. RESULT: A successful tension-free one-stage closure was obtained, without necrosis. Healing was uneventful without any complications and the coverage remained stable over 9 years with the child growth. CONCLUSION: The dorsal intercostal artery perforator propeller flap enables the surgeon to achieve a tension-free defect closure of considerably large myelomeningocele defects. This type of closure provides a durable coverage and a soft tissue padding over the neural tissues that persist with the child growth. The dorsal intercostal artery perforator propeller flap seems to be a useful closure of large myelomeningocele defects.
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Meningomielocele/cirugía , Colgajo Perforante , Procedimientos de Cirugía Plástica/métodos , Niño , Preescolar , Estudios de Seguimiento , Humanos , Lactante , Recién Nacido , Resultado del TratamientoRESUMEN
Large-conductance voltage- and calcium-activated K+ (BK) channels are key physiological players in muscle, nerve, and endocrine function by integrating intracellular Ca2+ and membrane voltage signals. The open probability of BK channels is regulated by the intracellular concentration of divalent cations sensed by a large structure in the BK channel called the "gating ring," which is formed by four tandems of regulator of conductance for K+ (RCK1 and RCK2) domains. In contrast to Ca2+ that binds to both RCK domains, Mg2+, Cd2+, or Ba2+ interact preferentially with either one or the other. Interaction of cations with their binding sites causes molecular rearrangements of the gating ring, but how these motions occur remains elusive. We have assessed the separate contributions of each RCK domain to the cation-induced gating-ring structural rearrangements, using patch-clamp fluorometry. Here we show that Mg2+ and Ba2+ selectively induce structural movement of the RCK2 domain, whereas Cd2+ causes motions of RCK1, in all cases substantially smaller than those elicited by Ca2+ By combining divalent species interacting with unique sites, we demonstrate that RCK1 and RCK2 domains move independently when their specific binding sites are occupied. Moreover, binding of chemically distinct cations to both RCK domains is additive, emulating the effect of fully occupied Ca2+ binding sites.
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Cationes Bivalentes/farmacología , Canales de Potasio de Gran Conductancia Activados por el Calcio/metabolismo , Canales de Potasio Calcio-Activados/metabolismo , Regulación Alostérica/fisiología , Secuencia de Aminoácidos , Animales , Sitios de Unión , Calcio/metabolismo , Proteínas de Unión al Calcio/metabolismo , Proteínas de Unión al Calcio/fisiología , Cationes/metabolismo , Cationes Bivalentes/metabolismo , Activación del Canal Iónico/fisiología , Subunidades alfa de los Canales de Potasio de Gran Conductancia Activados por Calcio/metabolismo , Potenciales de la Membrana/fisiología , Oocitos , Subunidades de Proteína , Xenopus laevis/embriologíaRESUMEN
The detection of IgG aquaporin-4 antibodies in the serum of patients with Neuromyelitis optica (NMO) has dramatically improved the diagnosis of this disease and its distinction from multiple sclerosis. Recently, a group of patients have been described who have an NMO spectrum disorder (NMOsd) and who are seronegative for AQP4 antibodies but positive for IgG aquaporin-1 (AQP1) or myelin oligodendrocyte glycoprotein (MOG) antibodies. The purpose of this study was to determine whether AQP1 and MOG could be considered new biomarkers of this disease; and if point mutations in the gDNA of AQP4, AQP1 and MOG genes could be associated with the etiology of NMOsd. We evaluated the diagnostic capability of ELISA and cell-based assays (CBA), and analyzed their reliability, specificity, and sensitivity in detecting antibodies against these three proteins. The results showed that both assays can recognize these antigen proteins under appropriate conditions, but only anti-AQP4 antibodies, and not AQP1 or MOG, appears to be a clear biomarker for NMOsd. CBA is the best method for detecting these antibodies; and serum levels of AQP4 antibodies do not correlate with the progression of this disease. So far, the sequencing analysis has not revealed a genetic basis for the etiology of NMOsd, but a more extensive analysis is required before definitive conclusions can be drawn.
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Anticuerpos/sangre , Acuaporina 1/genética , Acuaporina 4/genética , Glicoproteína Mielina-Oligodendrócito/genética , Neuromielitis Óptica/sangre , Neuromielitis Óptica/genética , Mutación Puntual/genética , Adulto , Biomarcadores/sangre , Estudios de Cohortes , Progresión de la Enfermedad , Femenino , Humanos , Masculino , Persona de Mediana EdadRESUMEN
Disabled-1 (Dab1) is an essential intracellular adaptor protein in the reelin pathway. Our previous studies in mice intestine showed that Dab1 transmits the reelin signal to cytosolic signalling pathways. Here, we determine the Dab1 isoform expressed in rodent small and large intestine, its subcellular location and co-localization with clathrin, caveolin-1 and N-Wasp. PCR and sequencing analysis reveal that rodent small and large intestine express a Dab1 isoform that misses three (Y198, Y200 and Y220) of the five tyrosine phosphorylation sites present in brain Dab1 isoform (canonical) and contains nuclear localization and export signals. Western blot assays show that both, crypts, which shelter progenitor cells, and enterocytes express the same Dab1 isoform, suggesting that epithelial cell differentiation does not regulate intestinal generation of alternatively spliced Dab1 variants. They also reveal that the canonical and the intestinal Dab1 isoforms differ in their total degree of phosphorylation. Immunostaining assays show that in enterocytes Dab1 localizes at the apical and lateral membranes, apical vesicles, close to adherens junctions and desmosomes, as well as in the nucleus; co-localizes with clathrin and with N-Wasp but not with caveolin-1, and in Caco-2 cells Dab1 localizes at cell-to-cell junctions by a Ca2+-dependent process. In conclusion, the results indicate that in rodent intestine a truncated Dab1 variant transmits the reelin signal and may play a role in clathrin-mediated apical endocytosis and in the control of cell-to-cell junction assembly. A function of intestinal Dab1 variant as a nucleocytoplasmic shuttling protein is also inferred from its sequence and nuclear location.
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Proteínas Adaptadoras Transductoras de Señales/metabolismo , Endocitosis , Uniones Intercelulares/metabolismo , Intestino Grueso/metabolismo , Intestino Delgado/metabolismo , Proteínas del Tejido Nervioso/metabolismo , Proteínas Adaptadoras Transductoras de Señales/genética , Animales , Células CACO-2 , Comunicación Celular/genética , Células Cultivadas , Endocitosis/genética , Expresión Génica , Humanos , Ratones , Ratones Endogámicos C57BL , Proteínas del Tejido Nervioso/genética , Unión Proteica , Isoformas de Proteínas , Ratas , Ratas Wistar , Proteína Reelina , Distribución TisularRESUMEN
Reelin is an extracellular matrix protein first known for its key role in neuronal migration. Studies in rodent small intestine suggested that reelin protects the organism from intestinal pathology. Here we determined in mice colon, by real time-PCR and immunological assays, the expression of the reelin signalling system; its response to dextran sulphate sodium (DSS) and the response of wild-type and reeler mice to DSS-treatment. DNA methylation was determined by bisulfite modification and sequencing of genomic DNA. In the colon mucosa reelin expression is restricted to the myofibroblasts, whereas both epithelial cells and myofibroblasts express reelin receptors (ApoER2 and VLDLR) and its effector protein Dab1. The muscle layer also expresses reelin. DSS-treatment reduces reelin expression in the muscle but it is activated in the mucosa. Activation of mucosal reelin is greater in magnitude and is delayed until after the activation of the myofibroblasts marker, α-SMA. This indicates that the DSS-induced reelin up-regulation results from changes in the reelin gene expression rather than from myofibroblasts proliferation. DSS-treatment does not modify Sp1 or Tbr1 mRNA abundance, but increases that of TGF-ß1 and ApoER2, decreases that of CASK and DNMT1 and it also decreases the reelin promoter methylation. Finally, the reeler mice exhibit higher inflammatory scores than wild-type mice, indicating that the mutation increases the susceptibility to DSS-colitis. In summary, this data are the first to demonstrate that mouse distal colon increases reelin production in response to DSS-colitis via a DNMT1-dependent hypo-methylation of the gene promoter region and that reelin provides protection against colitis.
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Moléculas de Adhesión Celular Neuronal/genética , Colitis/genética , Colon/metabolismo , Proteínas de la Matriz Extracelular/genética , Proteínas del Tejido Nervioso/genética , Serina Endopeptidasas/genética , Regulación hacia Arriba , Enfermedad Aguda , Animales , Colitis/inducido químicamente , Colitis/patología , Colon/patología , ADN (Citosina-5-)-Metiltransferasa 1/genética , Metilación de ADN , Sulfato de Dextran , Ratones Endogámicos C57BL , Miofibroblastos/metabolismo , Miofibroblastos/patología , Regiones Promotoras Genéticas , Proteína ReelinaRESUMEN
We previously reported that reelin, an extracellular matrix protein first known for its key role in neuronal migration, reduces the susceptibility to dextran sulphate sodium (DSS)-colitis. The aim of the current study was to determine whether reelin protects from colorectal cancer and how reelin defends from colon pathology. In the colon of wild-type and of mice lacking reelin (reeler mice) we have analysed the: i) epithelium cell renewal processes, ii) morphology, iii) Sox9, Cdx2, Smad5, Cyclin D1, IL-6 and IFNγ mRNA abundance in DSS-treated and untreated mice, and iv) development of azoxymethane/DSS-induced colorectal cancer, using histological and real time-PCR methodologies. The reeler mutation increases colitis-associated tumorigenesis, with increased tumours number and size. It also impairs the intestinal barrier because it reduces cell proliferation, migration, differentiation and apoptosis; decreases the number and maturation of goblet cells, and expands the intercellular space of the desmosomes. The intestinal barrier impairment might explain the increased susceptibility to colon pathology exhibited by the reeler mice and is at least mediated by the down-regulation of Sox9 and Cdx2. In response to DSS-colitis, the reeler colon increases the mRNA abundance of IL-6, Smad5 and Cyclin D1 and decreases that of IFNγ, conditions that might result in the increased colitis-associated tumorigenesis found in the reeler mice. In conclusion, the results highlight a role for reelin in maintaining intestinal epithelial cell homeostasis and providing resistance against colon pathology.
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Moléculas de Adhesión Celular Neuronal/metabolismo , Colitis/metabolismo , Colon/metabolismo , Enterocitos/metabolismo , Proteínas de la Matriz Extracelular/metabolismo , Regulación de la Expresión Génica , Proteínas del Tejido Nervioso/metabolismo , Proteínas Oncogénicas/biosíntesis , Serina Endopeptidasas/metabolismo , Animales , Apoptosis/efectos de los fármacos , Movimiento Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Colitis/inducido químicamente , Colitis/patología , Colon/patología , Sulfato de Dextran/toxicidad , Enterocitos/patología , Femenino , Masculino , Ratones , Proteína ReelinaRESUMEN
Reelin is an extracellular matrix protein that plays a critical role in neuronal migration. Here we show that the mucosa of human colon expresses reelin, its receptors ApoER2 and VLDLR, and its effector protein Dab1. Immunohistochemical analyses reveal that reelin expression is restricted to pericryptal myofibroblasts; Dab1 is detected at myofibroblasts, the apical domain of surface epithelial and crypt cells, and a strong linear staining is observed at the basement membrane; VLDLR and ApoER2 are in the cytoplasm of surface epithelium and myofibroblasts, and VLDLR is also detected in the cytoplasm of the crypt cells. Human colorectal cancer downregulates reelin without change in vimentin or N-cadherin mRNA levels. Decreased Reelin mRNA expression is accompanied by decreased HIC1 mRNA levels, increased mRNA levels of ApoER2 and DNMT1, increased reelin hypermethylation and no change in either Cask or TGF-ß1 mRNAs, suggesting that reelin repression results from a DNMT1-mediated hypermethylation of the reelin gene promoter. Decreased HIC1 expression may repress reelin transcription via increasing ApoER2 transcription. We conclude that the mucosa of human colon expresses the reelin-Dab1 signaling system and that reelin is repressed in colorectal cancer before epithelial-mesenchymal transition has occurred. The significant down-regulation of reelin expression makes this gene a promising biomarker for colorectal cancers. © 2016 Wiley Periodicals, Inc.
Asunto(s)
Proteínas Adaptadoras Transductoras de Señales/análisis , Moléculas de Adhesión Celular Neuronal/análisis , Colon/patología , Neoplasias Colorrectales/patología , Proteínas de la Matriz Extracelular/análisis , Mucosa Intestinal/patología , Proteínas del Tejido Nervioso/análisis , Recto/patología , Serina Endopeptidasas/análisis , Transducción de Señal , Proteínas Adaptadoras Transductoras de Señales/genética , Proteínas Adaptadoras Transductoras de Señales/metabolismo , Anciano , Anciano de 80 o más Años , Cadherinas/análisis , Cadherinas/genética , Cadherinas/metabolismo , Moléculas de Adhesión Celular Neuronal/genética , Moléculas de Adhesión Celular Neuronal/metabolismo , Colon/metabolismo , Neoplasias Colorrectales/genética , Neoplasias Colorrectales/metabolismo , Proteínas de la Matriz Extracelular/genética , Proteínas de la Matriz Extracelular/metabolismo , Femenino , Regulación Neoplásica de la Expresión Génica , Humanos , Mucosa Intestinal/metabolismo , Proteínas Relacionadas con Receptor de LDL/análisis , Proteínas Relacionadas con Receptor de LDL/genética , Proteínas Relacionadas con Receptor de LDL/metabolismo , Masculino , Persona de Mediana Edad , Proteínas del Tejido Nervioso/genética , Proteínas del Tejido Nervioso/metabolismo , Receptores de LDL/análisis , Receptores de LDL/genética , Receptores de LDL/metabolismo , Recto/metabolismo , Proteína Reelina , Serina Endopeptidasas/genética , Serina Endopeptidasas/metabolismoRESUMEN
UNLABELLED: Human immunodeficiency virus (HIV) replication is strongly dependent upon a programmed ribosomal frameshift. Here we investigate the relationships between the thermodynamic stability of the HIV type 1 (HIV-1) RNA frameshift site stem-loop, frameshift efficiency, and infectivity, using pseudotyped HIV-1 and HEK293T cells. The data reveal a strong correlation between frameshift efficiency and local, but not overall, RNA thermodynamic stability. Mutations that modestly increase the local stability of the frameshift site RNA stem-loop structure increase frameshift efficiency 2-fold to 3-fold in cells. Thus, frameshift efficiency is determined by the strength of the thermodynamic barrier encountered by the ribosome. These data agree with previous in vitro measurements, suggesting that there are no virus- or host-specific factors that modulate frameshifting. The data also indicate that there are no sequence-specific requirements for the frameshift site stem-loop. A linear correlation between Gag-polymerase (Gag-Pol) levels in cells and levels in virions supports the idea of a stochastic virion assembly mechanism. We further demonstrate that the surrounding genomic RNA secondary structure influences frameshift efficiency and that a mutation that commonly arises in response to protease inhibitor therapy creates a functional but inefficient secondary slippery site. Finally, HIV-1 mutants with enhanced frameshift efficiencies are significantly less infectious, suggesting that compounds that increase frameshift efficiency by as little as 2-fold may be effective at suppressing HIV-1 replication. IMPORTANCE: HIV, like many retroviruses, utilizes a -1 programmed ribosomal frameshift to generate viral enzymes in the form of a Gag-Pol polyprotein precursor. Thus, frameshifting is essential for viral replication. Here, we utilized a panel of mutant HIV strains to demonstrate that in cells, frameshifting efficiency is correlated with the stability of the local thermodynamic barrier to ribosomal translocation. Increasing the stability of the frameshift site RNA increases the frameshift efficiency 2-fold to 3-fold. Mutant viruses with increased frameshift efficiencies have significantly reduced infectivity. These data suggest that this effect might be exploited in the development of novel antiviral strategies.
Asunto(s)
Mutación del Sistema de Lectura/genética , Sistema de Lectura Ribosómico/genética , Proteínas de Fusión gag-pol/metabolismo , Infecciones por VIH/virología , VIH-1/genética , ARN Viral/genética , Virión/fisiología , Emparejamiento Base , Secuencia de Bases , Regulación Viral de la Expresión Génica , Células HEK293 , Infecciones por VIH/genética , VIH-1/química , VIH-1/metabolismo , Humanos , Conformación de Ácido Nucleico , Estabilidad del ARN , ARN Viral/química , ARN Viral/metabolismo , Ensamble de Virus , Replicación ViralRESUMEN
BACKGROUND: Traditionally, technical proficiency for spinal anesthesia has been assessed using observational scales such as global rating scales or task specific checklists. However more objective metrics are required in order to improve novice's training programs. The aim of this study is to validate the hand motion analysis of the Imperial College Surgical Assessment Device (ICSAD) in a simulated model of spinal anesthesia. METHODS: Three groups of physicians with different levels of experience were video recorded performing a spinal anesthesia in a simulated lumbar puncture torso. Participants' technical performance was assessed with ICSAD, a Global Rating Scale (GRS) and a specific Checklist. Differences between the 3 groups were determined by Kruskal-Wallis test with post hoc Dunn's correction for multiple comparisons. Spearman correlation coefficient between ICSAD variables and the scores of the observational scales were calculated to establish concurrent validity. RESULTS: Thirty subjects participated in the study: ten novice (first year residents), 10 intermediate (third year residents) and 10 experts (attending anesthesiologists). GRS scores were significantly higher in experts, than intermediates and novices. Regarding total path length, number of movements and procedural time measured with ICSAD, all groups had significant differences between them (p = 0.026, p = 0.045 and p = 0.005 respectively). Spearman correlation coefficient was -0,46 (p = 0.012) between total path length measured with ICSAD and GRS scores. CONCLUSIONS: This is the first validation study of ICSAD as an assessment tool for spinal anesthesia in a simulated model. Using ICSAD can discriminate proficiency between expert and novices and correlates with previously validated GRS. Its use in the assessment of spinal anesthesia proficiency provides complementary data to existing tools. Our results could be used to design future training programs with reliable goals to accomplish.