Estudi de l'evolució micromorfológica i funcional del trasplantament intestinal experimental

El objetivo de nuestro estudio fue evaluar si el fallo de la función intestinal absortiva observado después del trasplante de intestino delgado se debe a cambios en el tamaño de la superficie epitelial o bien proviene de un fallo en la función celular de los enterocitos. MATERIAL Y MÉTODOSSe realizaron trasplantes de intestino delgado (SBT) en ratas de acuerdo con la técnica de Monchik y Russell. Los animales fueron distribuidos en tres grupos (n=15 cada uno): Grupo A (control): asa simple de Thiry Vella; Grupo B: isotransplante heterotópico LEW-LEW; Grupo C: alotransplante heterotópico LBN-LEW. Se utilizó como solución de preservación Ringer lactato heparinizado a 4º C. Las ratas del grupo C se trataron con Ciclosporina (15mg/kg/24h IM). A los 21 dias del transplante heterotòpico, en 5 animales de cada grupo se realizó un segundo procedimiento quirúrgico para colocar el segmento de intestino transplantado en posición ortotópica. Mediante microscopía òptica y tinción H/E se observó la evolución micromorfológica de la mucosa intestinal, tomando muestras del intestino donante in situ antes de la perfusión y tras el transplante a los 7,14,21 y 36 dias. Estas muestras fueron procesadas mediante un sistema de analisis morfométrico de imagenes para quantificar cambios en el tamaño de las vellosidades en cuanto altura y anchura y asi determinar una posible modificacion en la superficie epitelial absortiva. En las series con transplante ortotópico, las muestras de intestino se estudiaron además por Microscopía Electrónica de Transmisión (TEM). Para determinar la evolución de la función absortiva del intestino transplantado se efectuaron pruebas de absorción de glucosa en los mismos intervalos de tiempo que las biopsias.RESULTADOS El estudio morfométrico muestra una disminución progresiva en la altura de las vellosidades tras el transplante, siendo más pronunciado en el grupo C. Una tendencia al aumento se observó en el ancho de las vellosidades. La superficie epitelial absortiva mostró una tendencia a la reducción, recuperando los valores iniciales tras la interposición ortotópica. Una reducción progresiva significativa de la absorción de glucosa se observó en ambos grupos de animales trasplantados respecto al grupo control. El estudio por TEM mostró la presencia de vacuolas citoplasmáticas en los enterocitos, así como una leve alteración en la morfología de las microvellosidades, mitocondrias y retículo endoplásmico. DISCUSIÓN Y CONCLUSIONES Una alteración de la fisiología celular parece ser la causa del fallo de la función de absorción intestinal después de SBT y este fracaso no dependería del tamaño de la superficie epitelial. Las alteraciones ultraestructurales observadas al TEM sugieren un daño celular grave que podría ser la causa de la insuficiencia absortiva. Pero el origen de estas alteraciones intracelulares sigue siendo desconocido pudiendo provenir tanto del efecto isquemia-reperfusión como de la respuesta inmunológica o de la toxicidad del propio tratamiento inmunosupresor.The aim of our study was to asses if the failure of the absortive intestinal function observed after small bowel transplantation is due either to changes in the size of epithelial surface or caused by a failure in the cellular function of enterocytes.MATERIALS AND METHODSSmall bowel transplants (SBT) were performed in rats according to Monchik and Russell's technique. Animals were distributed into three groups (n=15 each):Group A (control):simple Thiry-Vella loop; Group B:heterotopic isograft LEW-LEW; Group C:heterotopic allograft LBN-LEW. Heparinized lactated Ringer's at 4ºC was a cold preservation solution. Cyclosporine dose 15mg/kg/24h IM was administered to group C rats. At day 21 of the initial surgery, a second operative procedure was carried out on 5 of the transplanted animals of each group to place the transplanted small bowel in orthotopic position.To asses the micromorphology of intestinal mucosa by light microscopy (LM), biopsy specimens of the donor small bowel were taken in situ before perfusion and after transplant at 7,14,21 and 36 days. In those series with orthotopic transplantation bowel samples were studied, in addition, by Transmision Electron Microscopy (TEM).The absortive function of the transplanted bowel was observed by Glucose absorption test performed at same time points of the biopsies. Histomorphometric determinations of size of villus height and width, and total epithelial surface was performed by LM H/E and Image Processing and Analysis System.RESULTSMorphometrical study shows a progressive shortening of villus in both groups of transplanted animals, being more pronounced in the group C at the end of study. A tendency to increase was observed in the villus width. The absorptive epithelial surface showed an initial reduction followed of return to normal state after orthotopical interposition.A significative progressive reduction of glucose absorption was observed in both groups of transplanted animals than in the control group.Study by TEM showed cytoplasmic vacuoles in the absorptive cells. There was also a slight alteration on morphology of microvilli, mitochondries and endoplasmic reticulum.DISCUSSION/CONCLUSIONSAn alteration of cellular physiology underlies the failure of intestinal absorptive function after SBT and this failure does not depend on the size of epithelial surface. The findings of TEM suggest a severe ultrastructural damage could be the cause of cellular absorptive failure, but the cause of this cellular damage remains unkown

Soil aquifer treatment to remove priority organic pollutants in the Llobregat river area

The Llobregat River is the main source of water supply in this area. This river together with its aquifer has suffered from several damages which had contributed to endanger a suitable ecological and hydrological status; among them, pollution is a serious problem to deal with. In the last decades, the presence of organic pollutants in this river has been demonstrated [1,2]. Some of them are persistent to biological degradation and have shown to survive wastewater treatments almost unaltered and therefore they get surface waters where in some cases become recalcitrant pollutants. In order to mitigate the effect and presence of these compounds, Soil Aquifer Treatments (SAT) could be an approach since these treatments have demonstrated to be efficient to remove or decrease concentrations of such contaminants due to physico-chemical and biological processes occurring within the subsoil [3]. In the framework of the LIFE-ENSAT project, an organic substrate layer was installed in the basin of a recharge system located in the upper Llobregat delta aquifer. The aim was to enhance the degradation of contaminants through biochemical reactions on the reductive environment promoted in the unsaturated zone in order to improve the water quality of the system. In this work, the efficiency of this organic layer to remove organic pollutants included in European directives (2000/60/EC-2008/105/EC), such as triazines, pesticides or polyaromatic hydrocarbons (PAHs), has been evaluated. More than 80 compounds were analyzed; among them, 27 were priority pollutants while 6 were classified as preference substances (RD 60/2011).Peer ReviewedPostprint (published version

Flavonoid allosteric modulation of mutated visual rhodopsin associated with retinitis pigmentosa

Dietary flavonoids exhibit many biologically-relevant functions and can potentially have beneficial effects in the treatment of pathological conditions. In spite of its well known antioxidant properties, scarce structural information is available on the interaction of flavonoids with membrane receptors. Advances in the structural biology of a specific class of membrane receptors, the G protein-coupled receptors, have significantly increased our understanding of drug action and paved the way for developing improved therapeutic approaches. We have analyzed the effect of the flavonoid quercetin on the conformation, stability and function of the G protein-coupled receptor rhodopsin, and the G90V mutant associated with the retinal degenerative disease retinitis pigmentosa. By using a combination of experimental and computational methods, we suggest that quercetin can act as an allosteric modulator of opsin regenerated with 9-cis-retinal and more importantly, that this binding has a positive effect on the stability and conformational properties of the G90V mutant associated with retinitis pigmentosa. These results open new possibilities to use quercetin and other flavonoids, in combination with specific retinoids like 9-cis-retinal, for the treatment of retinal degeneration associated with retinitis pigmentosa. Moreover, the use of flavonoids as allosteric modulators may also be applicable to other members of the G protein-coupled receptors superfamily.Peer ReviewedPostprint (author's final draft

Sympathetic Hyperactivity and Sleep Disorders in Individuals With Type 2 Diabetes

Introduction: Many studies on the impact of type 2 diabetes mellitus (T2DM) on sleep breathing have shown a higher prevalence and severity of sleep apnea-hypopnea syndrome (SAHS) in those with T2DM. Moreover, an increased activity of the sympathetic nervous system has been described in both pathologies. This cross-sectional study aimed to assess sympathetic activity in patients with T2DM, and to investigate the relationship between sympathetic activity and polysomnographic parameters. Materials and Methods: Thirty-six patients with T2DM without known clinical macrovascularnorpulmonarydiseaseand11controlsunderwentrespiratorypolygraphy, and their cardiac variability and 24-h urine total metanephrines were measured. Results: SAHS was highly prevalent with a mean apnea-hypopnea index (AHI) in the range of moderate SAHS. In patients with T2DM, the nocturnal concentration of total metanephrines in urine were higher than diurnal levels [247.0 (120.0–1375.0) vs. 210.0 (92.0–670.0), p = 0.039]. The nocturnal total metanephrine concentration was positively and significantly associatedwith the percentage of sleeping time spent with oxygen saturation <90%(CT90). In the entire population and in subjects with T2DM, the multivariate regression analysis showed a direct interaction between the nocturnal concentration of urine metanephrines and the CT90. Conclusion: These findings suggest that the increase in sympathetic activity previously described in patients with T2DM could be mediated through nocturnal breathing disturbances. The diagnosis and treatment of SAHS may influence sympathetic activity disorders and may contribute to an improvement in T2DM and cardiovascular risk.This study was supported by grants from the Instituto de Salud Carlos III (Fondo de Investigación Sanitaria, PI12/00803 and PI15/00260), European Union (European Regional Development Fund, Fondo Europeo de Desarrollo Regional, Una manera de hacer Europa), the Fundación Sociedad Española Endocrinología y Nutrición, and Menarini Spain S.A. The funders had no role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript

Porcine Digestible Peptides (PDP) in Weanling Diets Regulates the Expression of Genes Involved in Gut Barrier Function, Immune Response and Nutrient Transport in Nursery Pigs

CRAG 10.3390/ani10122368This study was conducted to investigate the effects of dietary supplementation of porcine digestible peptides (PDP), spray-dried plasma (SDP), or a combination of both, on growth performance and the expression of genes related to intestinal function of weaned pigs. A total of 180 piglets (trial 1) and 198 piglets (trial 2) were used to evaluate the partial substitution of soybean ingredients with 2% SDP or 2% PDP (trial 1), and with 3% SDP or the combination of 1% SDP and 2% PDP (SDP-PDP; trial 2) during the pre-starter period (0-14 days). The gene expression of 56 genes was quantified in a qPCR platform in jejunum and ileum samples obtained from piglets 14 d after weaning (trial 2). Piglets fed SDP, PDP and SDP-PDP had a higher body weight (BW), average daily gain (ADG) and feed efficiency (G:F) than the soybean control on day 14 (p < 0.05). In addition, the combination of SDP and PDP upregulated ten genes in jejunum samples (p < 0.05) related to intestinal function. More research is needed to confirm that gene expression upregulation by PDP in combination with SDP has an impact on intestinal function and to elucidate its underlying mechanisms

Correlations between the NMR Lipoprotein Profile, APOE Genotype, and Cholesterol Efflux Capacity of Fasting Plasma from Cognitively Healthy Elderly Adults

Cholesterol efflux capacity (CEC) is of interest given its potential relationship with several important clinical conditions including Alzheimer's disease. The inactivation of the APOE locus in mouse models supports the idea that it is involved in determining the CEC. With that in mind, we examine the impact of the plasma metabolome profile and the APOE genotype on the CEC in cognitively healthy elderly subjects. The study subjects were 144 unrelated healthy individuals. The plasma CEC was determined by exposing cultured mouse macrophages treated with BODIPY-cholesterol to human plasma. The metabolome profile was determined using NMR techniques. Multiple regression was performed to identify the most important predictors of CEC, as well as the NMR features most strongly associated with the APOE genotype. Plasma 3-hydroxybutyrate was the variable most strongly correlated with the CEC (r = 0.365; p = 7.3 × 10-6). Male sex was associated with a stronger CEC (r = -0.326, p = 6.8 × 10-5). Most of the NMR particles associated with the CEC did not correlate with the APOE genotype. The NMR metabolomics results confirmed the APOE genotype to have a huge effect on the concentration of plasma lipoprotein particles as well as those of other molecules including omega-3 fatty acids. In conclusion, the CEC of human plasma was associated with ketone body concentration, sex, and (to a lesser extent) the other features of the plasma lipoprotein profile. The APOE genotype exerted only a weak effect on the CEC via the modulation of the lipoprotein profile. The APOE locus was associated with omega-3 fatty acid levels independent of the plasma cholesterol level.Peer reviewe

Social Representation of Dementia: An Analysis of 5,792 Consecutive Cases Evaluated in a Memory Clinic

Abstract. Background: Different interpretations of cognitive impairment and dementia due to differences in health structures, such as cultural differences could affect the diagnosis and treatment of the condition. it is reasonable to expect that the social and family impact of the disease and coping strategies will differ among societies. Objective: The general aim of this study is to understand the social representations of dementia, its associated practices, and the effects they imply. Methods: People diagnosed with clinical dementia and their families were assessed from 2005 to 2015 in the memory clinic of the Fundacio ́ ACE, Institut Catala` de Neurocie`ncies Aplicades in Barcelona, Spain. Results: 9,898 people were examined and 5,792 were diagnosed with dementia. For those with a caregiver (71%), the decision-making fell on the person with dementia in 16.2% of the cases; and for those without a caregiver, in 26.4% of the cases the family did not perceive the deficits as a disease, which led to multiple risk situations (74.6%). Conclusions: The recognition of dementia as part of aging is common among families. Consequently, risk situations may arise and diagnosis and access to treatment may be delayed. The incorporation of a social appraisal to the diagnostic process is a necessity to evaluate these situations

Dementia care in times of COVID-19: Experience at Fundació ACE in Barcelona, Spain

Background: Fundació ACE is a non-profit organization providing care based on a holistic model to persons with cognitive disorders and their families for 25 years in Barcelona, Spain. Delivering care to this vulnerable population amidst the COVID-19 pandemic has represented a major challenge to our institution. Objective: To share our experience in adapting our model of care to the new situation to ensure continuity of care. Methods: We detail the sequence of events and the actions taken within Fundació ACE to swiftly adapt our face-to-face model of care to one based on telemedicine consultations. We characterize individuals under follow-up by the Memory Unit from 2017 to 2019 and compare the number of weekly visits in 2020 performed before and after the lockdown was imposed. Results: The total number of individuals being actively followed by Fundació ACE Memory Unit grew from 6,928 in 2017 to 8,147 in 2019. Among those newly diagnosed in 2019, most patients had mild cognitive impairment or mild dementia (42% and 25%, respectively). Weekly visits dropped by 60% following the suspension of face-to-face activity. However, by April 24 we were able to perform 78% of the visits we averaged in the weeks before confinement began. Discussion: We have shown that Fundació ACE model of care has been able to successfully adapt to a health and social critical situation as COVID-19 pandemic. Overall, we were able to guarantee the continuity of care while preserving the safety of patients, families, and professionals. We also seized the opportunity to improve our model of care

Porcine Digestible Peptides (PDP) in Weanling Diets Regulates the Expression of Genes Involved in Gut Barrier Function, Immune Response and Nutrient Transport in Nursery Pigs

This study was conducted to investigate the effects of dietary supplementation of porcine digestible peptides (PDP), spray-dried plasma (SDP), or a combination of both, on growth performance and the expression of genes related to intestinal function of weaned pigs. A total of 180 piglets (trial 1) and 198 piglets (trial 2) were used to evaluate the partial substitution of soybean ingredients with 2% SDP or 2% PDP (trial 1), and with 3% SDP or the combination of 1% SDP and 2% PDP (SDP-PDP; trial 2) during the pre-starter period (0-14 days). The gene expression of 56 genes was quantified in a qPCR platform in jejunum and ileum samples obtained from piglets 14 d after weaning (trial 2). Piglets fed SDP, PDP and SDP-PDP had a higher body weight (BW), average daily gain (ADG) and feed efficiency (G:F) than the soybean control on day 14 (p < 0.05). In addition, the combination of SDP and PDP upregulated ten genes in jejunum samples (p < 0.05) related to intestinal function. More research is needed to confirm that gene expression upregulation by PDP in combination with SDP has an impact on intestinal function and to elucidate its underlying mechanisms