Inflammatory bowel disease induces pathological α-synuclein aggregation in the human gut and brain.

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.

COVID-19 Y su relación con factores sociodemográficos en la etapa pre-vacunatoria, provincia de Tucumán, Argentina, 2020-2021 / COVID-19 and its relationship with sociodemographic factors in the pre-vaccination period in Tucumán Province, Argentina, 2020-2021

RESUMEN INTRODUCCIÓN El impacto de COVID-19 en una población puede explicarse a través de los factores sociodemográficos y las respuestas de intervención. El objetivo fue evaluarlo en la provincia de Tucumán en la etapa previa a la vacunación. Se analizó la incidencia, letalidad y mortalidad de COVID-19 a nivel provincial y departamental, y se identificaron los factores sociodemográficos asociados. MÉTODOS Se realizó un estudio observacional de tipo ecológico con fuentes de datos secundarias en Tucumán. El período fue de marzo de 2020 a marzo de 2021. RESULTADOS Se registró una tasa de incidencia de COVID-19 de 4941 por 100 000 habitantes y una tasa de mortalidad de 93,29 por 100 000 habitantes. La incidencia de casos fue similar en varones y mujeres, y el grupo de 30 a 49 años presentó las tasas más altas. La tasa de mortalidad y letalidad en varones fue mayor que en mujeres, y en ambos sexos el grupo de 80 años o más presentó las mayores tasas. A nivel departamental, Capital, Tafí Viejo, Cruz Alta y Yerba Buena tuvieron las tasas de incidencia más altas. Burruyacú, Monteros y Trancas registraron las mayores tasas de mortalidad y letalidad. La incidencia de casos se vio afectada por la densidad poblacional y por el porcentaje de personas en hogares con alguna necesidad básica insatisfecha. DISCUSIÓN Se resalta la importancia de conocer la estructura, funcionalidad e identidad de las ciudades para comprender mejor su capacidad de resiliencia y adaptación frente a eventos como COVID-19.

ABSTRACT INTRODUCTION The population impact of COVID-19 can be explained by socio-demographic factors and the intervention responses. The objective was to evaluate it in the province of Tucumán in the pre-vaccination stage. Incidence, lethality and mortality of COVID-19 at province and department level were analyzed, identifying associated socio-demographic factors. METHODS An observational ecological study was conducted in Tucumán using secondary data sources. The period of study was from March 2020 to March 2021. RESULTS There was an incidence rate of COVID-19 of 4941 per 100000 inhabitants and a mortality rate of 93.29 per 100000 inhabitants. The incidence of cases was similar among males and females, and the age group between 30 and 49 years showed the highest rates. Mortality and lethality were higher in men than in women, and in both sexes the age group of 80 years and over presented the highest rates. At department level, Capital, Tafí Viejo, Cruz Alta and Yerba Buena had the highest incidence rates. Burruyacú, Monteros and Trancas had the highest mortality and lethality rates. The incidence of cases was affected by population density and by the percentage of people living in households with an unsatisfied basic need. DISCUSSION This work highlights the importance of knowing the structure, functionality and identity of cities to better understand their resilience and adaptation capacity in the face of events such as COVID-19.

Reelin Protects against Colon Pathology via p53 and May Be a Biomarker for Colon Cancer Progression.

Previous observations made in human and mouse colons suggest that reelin protects the colon from pathology. In this study, we evaluated reelin expression during the transition from either colitis or precancerous lesions to colon cancer and tried to elucidate reelin regulation under these transition processes. Samples of healthy and pathological colons from humans and mice treated with either azoxymethane/dextran sulfate sodium (DSS) or azoxymethane alone were used. The relative abundances of reelin, DNMT-1 and ApoER2 mRNAs were determined by PCR in the colon samples cited above and in the tissue adjacent to mouse colon polyps and adenocarcinomas. In both, humans and mice, reelin mRNA abundance increased significantly in ulcerative colitis and slightly in polyps and decreased in adenomas and adenocarcinomas. Reelin expression was higher in the tissue adjacent to the colon adenocarcinoma and lower in the lesion itself. The reelin expression changes may result, at least in part, from those in DNMT-1 and appear to be independent of ApoER2. Lack of reelin downregulated p-Akt and p53 in healthy colon and prevented their increases in the inflamed colon, whereas it increased GSK-3ß in DSS-untreated mice. In conclusion, reelin mRNA abundance depends on the severity of the colon pathology, and its upregulation in response to initial injuries might prevent the beginning of colon cancer, whereas reelin repression favors it. Increased p53 expression and activation may be involved in this protection. We also propose that changes in colon reelin abundance could be used to predict colon pathology progression.

Acute Colon Inflammation Triggers Primary Motor Cortex Glial Activation, Neuroinflammation, Neuronal Hyperexcitability, and Motor Coordination Deficits.

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.

The Role of Inflammation as a Preponderant Risk Factor in Cardiovascular Diseases.

Cardiovascular diseases cause considerable health and economic burden, as they are the leading cause of disability and death in the western world. Inactivity, hypertension, obesity, diabetes, and smoking are among the classic risk factors for cardiovascular disease. From a pathophysiological point of view, the arteries of our body bear the harmful stimuli produced by these factors and respond to them with a series of intricate adaptive mechanisms. Vascular remodeling constitutes an adaptive response to hemodynamic and inflammatory alterations associated with hypertension, diabetes, and other illnesses. Thickening of the arterial walls leads to endothelial dysfunction and increases the risk of cerebrovascular and coronary events. During the last decades, antiplatelet, lipid-lowering, and antihypertensive therapies have been the cornerstone of primary and secondary prevention of cardiovascular events. However, it is still unknown whether their efficacy is strictly associated with the control of the classical risk factors or their additive effects on vascular inflammation. Since inflammation of arterial walls is related to the pathogenesis of atherosclerosis, it has been hypothesized that anti-inflammatory therapies could prevent and treat vascular remodeling. Clinical trials based on canakinumab or hydroxychloroquine provide further insight into the role of inflammation in the pathophysiology of cardiovascular diseases. In this review, we have analyzed evidence and suggested that inflammation may play an important role in the final pathway of many cardiovascular risk factors.

Fecal Volatile Organic Compounds and Microbiota Associated with the Progression of Cognitive Impairment in Alzheimer's Disease.

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.

Galectin-3 Deletion Reduces LPS and Acute Colitis-Induced Pro-Inflammatory Microglial Activation in the Ventral Mesencephalon.

Parkinson's disease is a highly prevalent neurological disorder for which there is currently no cure. Therefore, the knowledge of risk factors as well as the development of new putative molecular targets is mandatory. In this sense, peripheral inflammation, especially the originated in the colon, is emerging as a predisposing factor for suffering this disease. We have largely studied the pleiotropic roles of galectin-3 in driving microglia-associated immune responses. However, studies aimed at elucidating the role of galectin-3 in peripheral inflammation in terms of microglia polarization are lacking. To achieve this, we have evaluated the effect of galectin-3 deletion in two different models of acute peripheral inflammation: intraperitoneal injection of lipopolysaccharide or gut inflammation induced by oral administration of dextran sodium sulfate. We found that under peripheral inflammation the number of microglial cells and the expression levels of pro-inflammatory mediators take place specifically in the dopaminergic system, thus supporting causative links between Parkinson's disease and peripheral inflammation. Absence of galectin-3 highly reduced neuroinflammation in both models, suggesting an important central regulatory role of galectin-3 in driving microglial activation provoked by the peripheral inflammation. Thus, modulation of galectin-3 function emerges as a promising strategy to minimize undesired microglia polarization states.

Asociación entre incidencia de varicela y variabilidad climática en la provincia de Tucumán, Argentina, 2005-2019 / Association between varicella incidence and climate variability in Tucumán province, Argentina, 2005-2019

INTRODUCTION: Varicella is a vaccine-preventable disease with marked seasonality. Few studies incorporate climatic variables to understand the epidemiological characteristics of this disease. The aim was to evaluate the relationship between varicella incidence and climatic variables in Tucumán (a province with temperate subtropical climate) during 2005-2019. METHODS: The relationship in pre- (2005-2014) and post-vaccination (2015-2019) periods was analyzed, identifying the associated climatic variables and the cut-off point where the risk of transmission increased. An observational ecological study was carried out with secondary data sources. R software was used. The information was split into three time series: 2005-2009, 2010-2014 and 2015-2019. For each period, a description of the time series was performed and generalized additive models (GAMs) were built using a negative binomial distribution. RESULTS: A seasonal behavior was observed, with peak incidence during spring in all periods. In the post-vaccination period, the peak occurred later (epidemiological week [EW] 46) than in the pre-vaccination periods (EW 43 and 42). Maximum temperature and relative humidity were associated during the first two periods, while minimum temperature, wind and thermal amplitude were associated in the third one. DISCUSSION: This study helped establish the relationship between climatic variables and varicella in Tucumán.

Proper E-cadherin membrane location in colon requires Dab2 and it modifies by inflammation and cancer.

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.

Análisis por series temporales de la incidencia de varicela y el impacto de la implementación de la vacuna en Tucumán / Time series analysis of the chickenpox rate and the impact of vaccine implementation in Tucumán

La varicela es una de las enfermedades inmunoprevenibles más comunes. En 1998 la Organización Mundial de la Salud recomendó incorporar la vacuna a los programas nacionales de vacunación. Argentina lo hizo en 2015. El objetivo de este trabajo fue analizar la serie temporal de varicela durante 2005-2019 y evaluar el impacto de la vacuna sobre su incidencia en Tucumán tras la implementación. MÉTODOS: Se llevó a cabo un estudio observacional de tipo ecológico con fuentes de datos secundarias. Los casos de varicela fueron los consignados por el Sistema Nacional de Vigilancia de Salud. Se describió la serie temporal de casos notificados de varicela para Tucumán y se construyeron modelos aditivos generalizados (GAM) utilizando una distribución binomial negativa. Se verificó el impacto de la vacuna tomando el período 2005-2014, se construyó un modelo GAM y se pronosticó el comportamiento más probable luego de la implementación. Se evaluó el impacto comparando las tasas con sus intervalos de confianza entre lo pronosticado y lo observado. RESULTADOS: Tucumán notificó 82 810 casos durante 2005-2019. La tasa anual varió entre 1,66 por 1000 habitantes (2019) y 6,04 por 1000 habitantes (2007). La serie presentó estacionalidad y en los últimos años una tendencia decreciente. Se observó una disminución significativa de la tasa de incidencia tras la implementación de la vacuna. DISCUSIÓN: El presente trabajo evidenció el impacto de una política pública como la vacuna

Uso de WhatsApp en neurocirugía: ¿problema o solución? / Use of WhatsApp in neurosurgery: problem or solution?

Introducción: Debido a la naturaleza impredecible y demandante de la neurocirugía, es necesaria una comunicación segura y rápida entre colegas. WhatsApp se presenta, en este contexto, como una solución rentable, fácil, accesible y rápida. Por la característica fugaz de la tecnología los avances preceden a las normativas. Actualmente WhatsApp es una herramienta frecuente en los equipos neuroquirúrgicos y nos encontramos utilizando una herramienta de forma diaria e ignorando las consecuencias que puede traer el intercambio de información de pacientes en un medio que no se encuentra diseñado para tal fin. El presente trabajo pretende medir la adhesión de WhatsApp en neurocirugía y el nivel de conocimiento sobre sus ventajas y desventajas. Métodos: Se elaboró una encuesta sobre el uso del WhatsApp en Neurocirugía que se entregó impresa aleatoriamente a 150 neurocirujanos. Resultados: Se encuestaron 112 neurocirujanos con una edad promedio de 34 años. Un 98% usa Smartphone y la totalidad de estos usa WhatsApp. Un 96% utiliza WhatsApp para resolver cuestiones médicas fuera de su horario de trabajo. Ninguno de los entrevistados conoce el marco legal de WhatsApp. Conclusiones: Existe una fuerte adhesión al uso de WhatsApp en la práctica médica por parte de los neurocirujanos en Argentina. Esto se debe a su uso fácil, rápido, grupal y económico, que se adecua a la perfección con el ritmo y complejidad de nuestra profesión. Sin embargo, existe un total desconocimiento del marco legal de esta aplicación, del déficit de seguridad de los datos on-line; lo que se traduce en un potencial riesgo a la confidencialidad del acto médico.

Background: Due to the unpredictable and demanding nature of neurosurgery, safe and rapid communication between colleagues is necessary. WhatsApp is presented as a cost-effective, easy and accessible solution. Because of the rapid advanced of technology, it usually precedes regulations. Currently WhatsApp is a frequent tool in neurosurgical teams and we find ourselves using an instrument on a daily basis and ignoring the consequences that the exchange of patient information using this method can bring. The present work aims to measure the adherence of WhatsApp in neurosurgery and the level of knowledge about its advantages and disadvantages. Methods: A survey was developed on the use of WhatsApp in Neurosurgery that was delivered randomly to 150 neurosurgeons. Results: 112 neurosurgeons with an average age of 34 years were surveyed. 98% use Smartphone and all of them use WhatsApp. 96% use WhatsApp to resolve medical issues outside of their work hours. None of the interviewees knows the legal framework of WhatsApp. Conclusions: There is a strong adherence to the use of WhatsApp in medical practice by neurosurgeons in Argentina. This is due to its easy, fast, and economic use, which is perfectly adapted to the rhythm and complexity of our profession. However, there is a total ignorance of the legal framework of this application which translates into potential risks to patient's confidentiality.

Two-Year Outcome of Aflibercept for the Treatment of Choroidal Neovascularization in Punctate Inner Choroidopathy.

Punctate inner choroidopathy (PIC) is a rare inflammatory chorioretinopathy that predominantly affects young myopic women. Visual prognosis is generally good, but occurrence of choroidal neovascularization (CNV) is common and may be vision threatening. Case reports and short case series support the effectiveness of intravitreal anti-vascular endothelial growth factor (VEGF) agents (ranibizumab and bevacizumab) for CNV associated with PIC given their anti-angiogenic and anti-inflammatory effects. Evidence concerning aflibercept, a more recent intravitreal anti-VEGF, is limited to a single case report. In this case report, we illustrate the case of a 43-year-old myopic woman presenting with visual acuity loss and distortion in the right eye over the last 5 days in whom CNV associated with PIC was diagnosed. Treatment with 1 injection per month of intravitreal aflibercept for 2 months and full-dose oral prednisone for 1 week, being tapered afterwards, improved visual acuity and resolved CNV, with benefits lasting up to 24 months.

Impact of diuretics on the urate lowering therapy in patients with gout: analysis of an inception cohort.

BACKGROUND: Diuretics have been associated with impaired response and refractoriness in gout, but whether this effect is still present with new urate-lowering drugs (ULD) and treat-to-target strategies is unknown. The aim of the present study was to assess the impact of the diuretics on the response to ULD in patients with gout.  METHODS: This was a retrospective analysis of an inception cohort. Participants were classified according to the type of ULD prescribed. We analysed the maximal dose of ULD (primary outcome variable), serum urate (SU) reduction, and the achievement of different SU targets (6 mg/dL, 5 mg/dL, and 4 mg/dL), according to the type of ULD prescribed and use of diuretics (loop and/or thiazide). We adjusted for confounders using multiple linear regression analysis. RESULTS: We included 245 patients: 208 treated with allopurinol (66 on diuretics, 31.7%), 35 with febuxostat (19 on diuretics, 57.6%), and 2 with benzbromarone. Significantly fewer participants in the allopurinol plus diuretics subgroup achieved SU levels of less than 5 mg/dL, but we found no other significant differences in SU targets associated with diuretics. Regarding the maximum ULD dose, a simple linear regression suggested an inverse relationship with diuretics (beta = - 0.125, p = 0.073), but this did not hold in the multivariable analysis (beta = - 0.47, p = 0.833). There was no association with febuxostat (beta = - 0.116, p = 0.514). CONCLUSION: Diuretics do not appear to have a significant impact on managing gout.

Small and large intestine express a truncated Dab1 isoform that assembles in cell-cell junctions and co-localizes with proteins involved in endocytosis.

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.

High fat diet-induced metabolically obese and normal weight rabbit model shows early vascular dysfunction: mechanisms involved.

BACKGROUND: Obesity contributes significantly to the development and evolution of cardiovascular disease (CVD) which is believed to be mediated by oxidative stress, inflammation and endothelial dysfunction. However, the vascular health of metabolically obese and normal weight (MONW) individuals is not completely comprehended. OBJECTIVES: The purpose of our study was to evaluate vascular function on the basis of a high fat diet (HFD)-MONW rabbit model. SUBJECTS: Twenty four male rabbits were randomly assigned to receive either a regular diet (CD, n = 12) or a high-fat diet (18% extra fat on the regular diet, HFD, n = 12) for 6 weeks. RESULTS: Body weight, TBARS and gluthathione serum levels were similar between the groups; fasting glucose, triglycerides, C reactive protein (CRP), visceral adipose tissue (VAT), triglyceride-glucose index (TyG index) were higher in the HFD group. Compared to CD, the HFD rabbits had glucose intolerance and lower HDL-cholesterol and plasma nitrites levels. Thoracic aortic rings from HFD rabbits exhibited: (a) a reduced acetylcholine-induced vasorelaxation; (b) a greater contractile response to norepinephrine and KCl; (c) an improved angiotensin II-sensibility. The HFD-effect on acetylcholine-response was reversed by the cyclooxygenase-2 (COX-2) inhibitor (NS398) and the cyclooxygenase-1 inhibitor (SC560), and the HFD-effect on angiotensin II was reversed by NS398 and the TP receptor blocker (SQ29538). Immunohistochemistry and western blot studies showed COX-2 expression only in arteries from HFD rabbits. CONCLUSIONS: Our study shows a positive pro-inflammatory status of HFD-induced MONW characterized by raised COX-2 expression, increase of the CRP levels, reduction of NO release and oxidative stress-controlled conditions in an early stage of metabolic alterations characteristic of metabolic syndrome. Endothelial dysfunction and increased vascular reactivity in MONW individuals may be biomarkers of early vascular injury. Therefore, the metabolic changes induced by HFD even in normal weight individuals may be associated to functional alterations of blood vessels.

MicroRNA MIMIC binding sites: Minor flanking nucleotide alterations can strongly impact MIMIC silencing efficacy in Arabidopsis.

In plants, microRNA (miRNA) target MIMICs (MIMs) have been widely used to inhibit miRNA function. They are based on the Arabidopsis INSENSITIVE TO PHOSPATE STARVATION 1 (IPS1) gene that corresponds to a non-coding RNA containing a miR399 binding site that can be modified to sequester and inhibit any miRNA of interest. However, the efficacy of miRNA inhibition of these different MIMs can vary greatly. Using MIMs that have strong efficacy (MIM159) and poor efficacy (MIM165), we investigate the underlying cause of this variation. Firstly, sequence alignments of IPS1 homologs from the Brassicaceae identified a highly conserved sequence immediately downstream of the miRNA binding site. Mutating this sequence in the context of the MIM159 attenuates its strong efficacy. This conserved flanking region contains a predicted stem-loop structure that is also predicted to be present in most modified MIMs that appear to have a strong efficacy, but not in MIM165 that has a poor efficacy. Restoring this predicted stem-loop in MIM165 via mutation of only three or five nucleotides within the conserved flanking region resulted in MIM165 variants that have very strong efficacies of miRNA inhibition. However, specifically mutating this predicted stem-loop in the MIM159 context failed to significantly reduce efficacy, and additional mutations to restore this predicted stem-loop weakened efficacy further. Although this shows there is no simple correlation between this predicted stem-loop and efficacy, these results add to the growing evidence that the sequence context of miRNA binding sites is important, and that minor nucleotide substitutions to flanking sequences of miRNA binding sites can strongly enhance or attenuate the miRNA-target interaction.

Early Discharge Programme on Hospital-at-Home Evaluation for Patients with Immediate Postoperative Course after Laparoscopic Colorectal Surgery.

BACKGROUND: To audit the safety of the early hospital discharge care model offered by a Hospital-at-home (HAH) unit during early postoperative follow-up of these patients, and to determine whether this care model is more efficient compared to the traditional care model. METHODS: A prospective study of 50 patients included consecutively for 1 year in an early discharge programme after laparoscopic colorectal surgery was performed. As of day 3 after surgery, if the patient met the relevant inclusion criteria they were transferred to the HAH unit. The domiciliary protocol consists of daily clinical follow-up and a series of analytical controls with the purpose of early detection of postoperative complications. If the clinical course was favourable on day 7 after the postoperative period the patient was discharged. RESULTS: A total of 66% were males, and the mean age was 60.6 years. The surgical procedure most commonly performed was sigmoidectomy. The mean stay was 5.5 days. There were no deaths during follow-up. The average estimated cost per day of stay in a HAH system was EUR 174.29 whilst the same average cost on a surgery ward stood at EUR 1,032.42. CONCLUSIONS: For patients undergoing major colorectal surgery with minimally invasive surgical technique, an early hospital discharge care programme by means of referral to a HAH unit is a safe and efficient care model which entails a significant cost saving for the public healthcare system.

Reelin protects from colon pathology by maintaining the intestinal barrier integrity and repressing tumorigenic genes.

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.

Reelin expression is up-regulated in mice colon in response to acute colitis and provides resistance against colitis.

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.