Immunomodulatory properties of the protein fraction from Phorphyra columbina

The phycobiliproteins from Rhodophyta, R-phycoerythrin (R-PE) and C-phycocyanin (C-PC), have been shown to exert immunomodulatory effects. This study evaluated the effects of a Phorphyra columbina protein fraction (PF) and R-PE and C-PC on rat primary splenocytes, macrophages, and T-lymphocytes in vitro. PF featured various protein species, including R-PE and C-PC. PF showed mitogenic effects on rat splenocytes and was nontoxic to cells except at 1 g L-1 protein. IL-10 secretion was enhanced by PF in rat splenocytes, macrophages, and especially T-lymphocytes, whereas it was markedly diminished by R-PE and C-PC. The production of pro-inflammatory cytokines by macrophages was inhibited. The effect of PF on IL-10 was evoked by JNK/p38 MAPK and NF-κB-dependent pathways in macrophages and T-lymphocytes. It was concluded that PF has immunomodulatory effects on macrophages and lymphocytes that appear to be predominantly anti-inflammatory via up-regulated IL-10 production and cannot be accounted for by R-PE and C-PC.Fil: Cian, Raúl Esteban. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación de Agroindustria. Instituto de Tecnología de Alimentos; Argentina. Universidad Nacional del Litoral. Facultad de Ingeniería Química; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: López Posadas, Rocío. Universidad de Granada; EspañaFil: Drago, Silvina Rosa. Universidad Nacional del Litoral. Facultad de Ingeniería Química; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Sánchez De Medina, Fermín. Universidad de Granada; EspañaFil: Martínez Augustin, Olga. Universidad de Granada; Españ

Efficacy and Safety of a Novel Submucosal Injection Solution for Resection of Gastrointestinal Lesions

Endoscopic mucosal resection (EMR) and endoscopic submucosal dissection (ESD) are minimally invasive and efficient techniques for the removal of gastrointestinal (GI) mucosal polyps. In both techniques, submucosal injection solutions are necessary for complete effectiveness and safety during the intervention to be obtained. The main objective of this study was to evaluate the efficacy and safety of a new sterile submucosal injection solution for EMR/ESD used within a clinical protocol in patients with intestinal polyps. We carried out a prospective study between 2016 and 2017 with patients who attended the Endoscopy Consultation—Digestive Department of Primary Hospital. Patients were selected for EMR/ESD after the application of clinical protocols. Thirty-six patients were selected (≥ 66 years with comorbidities and risk factors). Lesions were located mainly in the colon. Our solution presented an intestinal lift ≥ 60 min in EMR/ESD and a high expansion of tissue, optimum viscosity, and subsequent complete resorption. The genes S100A9 and TP53 presented an expression increase in the distal regions. TP53 and PCNA were the only genes whose expression was increased in polyp specimens vs. the surrounding tissue at the mRNA level. In EMR/ESD, our solution presented a prolonged effect at the intestinal level during all times of the intervention. Thus, our solution seems be an effective and safe alternative in cases of flat lesions in both techniques.Study co-financed by the Junta de Andalucia (PIN-0479-2016, CTS676, CTS235, CTS164), the Ministry of Economy and Competitivity, Spain (SAF2017-88457-R, AGL2017-85270-R), Nakafarma S.L and CIBERehd is funded by Instituto de Salud Carlos III, Spain. The sponsors had no role in the design, execution, interpretation, or writing of the study

Papel de la enzima fosfatasa alcalina no específica de tejido (TNAP) en el epitelio intestinal en la inflamación

Introducción: la fosfatasa alcalina (AP) es una familia de enzimas que ha sido relacionada con la protección frente a inflamación intestinal. Se ha descrito que una de sus isoformas, la fosfatasa alcalina intestinal (IAP), es capaz de desfosforilar diferentes antígenos bacterianos, de tal forma que la enzima regula el crecimiento de la microbiota e impide el paso de antígenos activos. En cuanto a la isoforma TNAP, se ha observado que su expresión se encuentra incrementada en la colitis experimental, no solo debido a la infiltración de células del sistema inmunológico, sino también por el incremento de expresión de esta enzima en las células del epitelio intestinal. Objetivo: conocer el papel de la TNAP en la inflamación intestinal. Métodos y resultados: se ha generado un modelo de ratón con deleción condicional inducible del gen que codifica TNAP (Alpl) en el epitelio intestinal (ratones AlplIEC-/-). El silenciamiento específico de TNAP en IECs en inflamación por DSS (7 días) supuso una pérdida mayor de peso en los ratones, sin observarse diferencias en el índice de actividad de la enfermedad (DAI). A nivel histológico se observó un mayor nivel de infiltración en la submucosa en el colon de los ratones sin TNAP. Los ratones AlplIEC-/- presentaron una expresión reducida de marcadores inflamatorios en el colon, como S100a8, Il6 y Tnf. Por el contrario, la deficiencia en TNAP en el epitelio intestinal supuso un aumento en la expresión de la fosfatasa alcalina intestinal global (Akp6) en el colon, sugiriendo que podría existir algún mecanismo de compensación. Además, la ausencia de TNAP en el epitelio intestinal provocó un aumento de expresión de genes relacionados en el mantenimiento de la función de barreara, como Muc4, Tjp1 y Tff3. Conclusión: los ratones AlplIEC-/- presentan un fenotipo mixto, con mayor infiltración y daño histológico pero menor expresión de marcadores inflamatorios en la colitis por DSS. Perspectivas futuras: se realizarán estudios de transcriptómica para conocer el efecto de la TNAP presente en el intestino sobre la función de barrera intestinal y sobre la microbiota

Fructooligosacharides Reduce Pseudomonas aeruginosa PAO1 Pathogenicity through Distinct Mechanisms

Pseudomonas aeruginosa is ubiquitously present in the environment and acts as an opportunistic pathogen on humans, animals and plants. We report here the effects of the prebiotic polysaccharide inulin and its hydrolysed form FOS on this bacterium. FOS was found to inhibit bacterial growth of strain PAO1, while inulin did not affect growth rate or yield in a significant manner. Inulin stimulated biofilm formation, whereas a dramatic reduction of the biofilm formation was observed in the presence of FOS. Similar opposing effects were observed for bacterial motility, where FOS inhibited the swarming and twitching behaviour whereas inulin caused its stimulation. In co-cultures with eukaryotic cells (macrophages) FOS and, to a lesser extent, inulin reduced the secretion of the inflammatory cytokines IL-6, IL-10 and TNF- a . Western blot experiments indicated that the effects mediated by FOS in macrophages are associated with a decreased activation of the NF- k B pathway. Since FOS and inulin stimulate pathway activation in the absence of bacteria, the FOS mediated effect is likely to be of indirect nature, such as via a reduction of bacterial virulence. Further, this modulatory effect is observed also with the highly virulent ptxS mutated strain. Co-culture experiments of P. aeruginosa with IEC18 eukaryotic cells showed that FOS reduces the concentration of the major virulence factor, exotoxin A, suggesting that this is a possible mechanism for the reduction of pathogenicity. The potential of these compounds as components of antibacterial and anti-inflammatory cocktails is discussed.The authors acknowledge financial support from FEDER funds and Fondo Social Europeo through grants from the Spanish Ministry of Economy and Competitiveness (grants SAF2011-22922, SAF2011-22812) the Andalusian regional government Junta de Andalucía (grant CVI-7335) and the Centre of Networked Biomedical Research on Hepatic and Digestive Diseases (CIBERehd) which is funded by the Carlos III Health Institute and the Ramón Areces Foundation, Spain

Reducing the environmental impact of surgery on a global scale: systematic review and co-prioritization with healthcare workers in 132 countries

Abstract Background Healthcare cannot achieve net-zero carbon without addressing operating theatres. The aim of this study was to prioritize feasible interventions to reduce the environmental impact of operating theatres. Methods This study adopted a four-phase Delphi consensus co-prioritization methodology. In phase 1, a systematic review of published interventions and global consultation of perioperative healthcare professionals were used to longlist interventions. In phase 2, iterative thematic analysis consolidated comparable interventions into a shortlist. In phase 3, the shortlist was co-prioritized based on patient and clinician views on acceptability, feasibility, and safety. In phase 4, ranked lists of interventions were presented by their relevance to high-income countries and low–middle-income countries. Results In phase 1, 43 interventions were identified, which had low uptake in practice according to 3042 professionals globally. In phase 2, a shortlist of 15 intervention domains was generated. In phase 3, interventions were deemed acceptable for more than 90 per cent of patients except for reducing general anaesthesia (84 per cent) and re-sterilization of ‘single-use’ consumables (86 per cent). In phase 4, the top three shortlisted interventions for high-income countries were: introducing recycling; reducing use of anaesthetic gases; and appropriate clinical waste processing. In phase 4, the top three shortlisted interventions for low–middle-income countries were: introducing reusable surgical devices; reducing use of consumables; and reducing the use of general anaesthesia. Conclusion This is a step toward environmentally sustainable operating environments with actionable interventions applicable to both high– and low–middle–income countries

Disturbances in metabolic, transport and structural genes in experimental colonic inflammation in the rat: a longitudinal genomic analysis

Abstract Background Trinitrobenzenesulphonic acid (TNBS) induced rat colitis is one of the most widely used models of inflammatory bowel disease (IBD), a condition whose aetiology and pathophysiology are incompletely understood. We have characterized this model at the genomic level using a longitudinal approach. Six control rats were compared with colitic animals at 2, 5, 7 and 14 days after TNBS administration (n = 3). The Affymetrix Rat Expression Array 230 2.0 system was used. Results TNBS-induced colitis had a profound impact on the gene expression profile, which was maximal 5 and 7 days post-induction. Most genes were affected at more than one time point. They were related to a number of biological functions, not only inflammation/immunity but also transport, metabolism, signal transduction, tissue remodeling and angiogenesis. Gene changes generally correlated with the severity of colitis. The results were successfully validated in a subset of genes by real-time PCR. Conclusion The TNBS model of rat colitis has been described in detail at the transcriptome level. The changes observed correlate with pathophysiological disturbances such as tissue remodelling and alterations in ion transport, which are characteristic of both this model and IBD.</p

Experimental acute pancreatitis is enhanced in mice with tissue nonspecific alkaline phoshatase haplodeficiency due to modulation of neutrophils and acinar cells.

Tissue nonspecific alkaline phosphatase (TNAP) has a well established role in bone homeostasis and in hepatic/biliary conditions. In addition, TNAP is expressed in the inflamed intestine and is relevant to T and B lymphocyte function. TNAP KO mice are only viable for a few days, but TNAP+/- haplodeficient mice are viable. Acute pancreatitis was induced by repeated caerulein injection in WT and TNAP+/- mice. TNAP+/- mice presented an increased expression of Cxcl2, Ccl2, Selplg (P-selectin ligand), Il6 and Il1b in the pancreas. Freshly isolated acinar cells showed a dramatic upregulation of Cxcl1, Cxcl2, Ccl2, Il6, Selpg or Bax in both pancreatitis groups. TNAP+/- cells displayed a 2-fold higher expression of Cxcl2, and a smaller increase in Il6. These findings could be partly replicated by in vitro treatment of primary acinar cells with caerulein. Furthermore, the proinflammatory effect on acinar cells could be partially reproduced in wild type cells treated with the TNAP inhibitor levamisole. TNAP mRNA levels were also markedly upregulated by pancreatitis in acinar cells. Neutrophil infiltration (MRP8+ cells) and activation (IL-6 and TNF production in LPS treated primary neutrophils) were increased in TNAP+/- vs WT mice. Neutrophil depletion greatly attenuated inflammation, indicating that this cell type is mainly responsible for the higher inflammatory status of TNAP+/- mice. In conclusion, our results show that altered TNAP expression results in heightened pancreatic inflammation, which may be explained by an augmented response of neutrophils and by a higher sensitivity of acinar cells to caerulein injury

Papel biológico de genes ubicados en una isla genómica en Pseudomonas aeruginosa

Pseudomonas aeruginosa es un patógeno humano oportunista Gram negativo omnipresente que no se considera parte de la microbiota intestinal comensal humana. Sin embargo, el agotamiento de la microbiota intestinal (disbiosis) después del tratamiento con antibióticos facilita la colonización del tracto intestinal por P. aeruginosa resistente a múltiples fármacos. Una posible alternativa a los tratamientos convencionales se basa en el uso de alimentos funcionales con actividad prebiótica. El efecto bifidogénico de fructooligosacáridos (FOS) está bien establecido, se ha demostrado que promueve el crecimiento de bacterias intestinales beneficiosas específicas como las bifidobacterias. Estudios previos del patógeno oportunista nosocomial Pseudomonas aeruginosa PAO1 han demostrado que los FOS reducen el crecimiento y la formación de biopelículas, debido a una disminución en la motilidad y la secreción de exotoxina. Sin embargo, la base transcripcional de estas alteraciones fenotípicas sigue sin estar clara. Para abordar esta pregunta, se realizó un análisis de secuencia de ARN (RNAseq) que nos permitió detectar la presencia de una isla genómica formada por 15 genes que fueron reprimidos en presencia de FOS. Anteriormente, se demostró mediante el análisis funcional de mutantes isogénicos, que los genes PA0643, PA0644 y PA0646, ubicados en esta isla génica, codifican proteínas involucradas en el crecimiento, la formación de biopelículas, la motilidad. En este contexto, este trabajo plasmo la implicación de estos genes en la modulación de la respuesta inflamatoria