NOD1 downregulates intestinal serotonin transporter and interacts with other pattern recognition receptors

Serotonin (5-HT) is an essential gastrointestinal modulator whose effects regulate the intestinal physiology. 5-HT effects depend on extracellular 5-HT bioavailability, which is controlled by the serotonin transporter (SERT) expressed in both the apical and basolateral membranes of enterocytes. SERT is a critical target for regulating 5-HT levels and consequently, modulating the intestinal physiology. The deregulation of innate immune receptors has been extensively studied in inflammatory bowel diseases (IBD), where an exacerbated defense response to commensal microbiota is observed. Interestingly, many innate immune receptors seem to affect the serotonergic system, demonstrating a new way in which microbiota could modulate the intestinal physiology. Therefore, our aim was to analyze the effects of NOD1 activation on SERT function, as well as NOD1's interaction with other immune receptors such as TLR2 and TLR4. Our results showed that NOD1 activation inhibits SERT activity and expression in Caco-2/TC7 cells through the extracellular signal-regulated kinase (ERK) signaling pathway. A negative feedback between 5-HT and NOD1 expression was also described. The results showed that TLR2 and TLR4 activation seems to regulate NOD1 expression in Caco-2/TC7 cells. To assess the extend of cross-talk between NOD1 and TLRs, NOD1 expression was measured in the intestinal tract (ileum and colon) of wild type mice and mice with individual knockouts of TLR2, and TLR4 as well as double knockout TLR2/TLR4 mice. Hence, we demonstrate that NOD1 acts on the serotonergic system decreasing SERT activity and molecular expression. Additionally, NOD1 expression seems to be modulated by 5-HT and other immune receptors as TLR2 and TLR4. This study could clarify the relation between both the intestinal serotonergic system and innate immune system, and their implications in intestinal inflammation

Modelling evaporation in electron‑beam physical vapour deposition of thermal barrier coatings

This work presents computational models of ingot evaporation for electron-beam physical vapour deposition (EB-PVD) that can be applied to the deposition and development of thermal barrier coatings (TBCs). TBCs are insulating coatings that protect aero-engine components from high temperatures, which can be above the component’s melting point. The development of advanced TBCs is fuelled by the need to improve engine efficiency by increasing the engine operating temperature. Rare-earth zirconates (REZ) have been proposed as the next-generation TBCs due to their low coefficient of thermal conductivity and resistance to molten calcium-magnesium alumina-silicates (CMAS). However, the evaporation of REZ has proven to be challenging, with some coatings displaying compositional segregation across their thickness. The computational models form part of a larger analytical model that spans the whole EB-PVD process. The computational models focus on ingot evaporation, have been implemented in MATLAB and include data from 6 oxides: ZrO2, Y2O3, Gd2O3, Er2O3, La2O3 and Yb2O3. Two models (2D and 3D) successfully evaluate the evaporation rates of constituent oxides from multiple-REZ ingots, which can be used to highlight incompatibilities and preferential evaporation of some of these oxides. A third model (local composition activated, LCA) successfully predicts the evaporation rate of the whole ingot and replicates the cyclic change in composition of the evaporated plume, which is manifested as changes in compositional segregation across the coating’s thickness. The models have been validated with experimental data from Cranfield University’s EB-PVD coaters, published vapour pressure calculations and evaporation rate formulas described in the literature

Systemic Exosomal Delivery of shRNA Minicircles Prevents Parkinsonian Pathology

The development of new therapies to slow down or halt the progression of Parkinson's disease is a health care priority. A key pathological feature is the presence of alpha-synuclein aggregates, and there is increasing evidence that alpha-synuclein propagation plays a central role in disease progression. Consequently, the downregulation of alpha-synuclein is a potential therapeutic target. As a chronic disease, the ideal treatment will be minimally invasive and effective in the long-term. Knockdown of gene expression has clear potential, and siRNAs specific to alpha-synuclein have been designed; however, the efficacy of siRNA treatment is limited by its short-term efficacy. To combat this, we designed shRNA minicircles (shRNA-MCs), with the potential for prolonged effectiveness, and used RVG-exosomes as the vehicle for specific delivery into the brain. We optimized this system using transgenic mice expressing GFP and demonstrated its ability to downregulate GFP protein expression in the brain for up to 6 weeks. RVG-exosomes were used to deliver anti-alpha-synuclein shRNA-MC therapy to the alpha-synuclein preformed-fibril-induced model of parkinsonism. This therapy decreased alpha-synuclein aggregation, reduced the loss of dopaminergic neurons, and improved the clinical symptoms. Our results confirm the therapeutic potential of shRNA-MCs delivered by RVG-exosomes for long-term treatment of neurodegenerative diseases

Global overview of the management of acute cholecystitis during the COVID-19 pandemic (CHOLECOVID study)

Background: This study provides a global overview of the management of patients with acute cholecystitis during the initial phase of the COVID-19 pandemic. Methods: CHOLECOVID is an international, multicentre, observational comparative study of patients admitted to hospital with acute cholecystitis during the COVID-19 pandemic. Data on management were collected for a 2-month study interval coincident with the WHO declaration of the SARS-CoV-2 pandemic and compared with an equivalent pre-pandemic time interval. Mediation analysis examined the influence of SARS-COV-2 infection on 30-day mortality. Results: This study collected data on 9783 patients with acute cholecystitis admitted to 247 hospitals across the world. The pandemic was associated with reduced availability of surgical workforce and operating facilities globally, a significant shift to worse severity of disease, and increased use of conservative management. There was a reduction (both absolute and proportionate) in the number of patients undergoing cholecystectomy from 3095 patients (56.2 per cent) pre-pandemic to 1998 patients (46.2 per cent) during the pandemic but there was no difference in 30-day all-cause mortality after cholecystectomy comparing the pre-pandemic interval with the pandemic (13 patients (0.4 per cent) pre-pandemic to 13 patients (0.6 per cent) pandemic; P = 0.355). In mediation analysis, an admission with acute cholecystitis during the pandemic was associated with a non-significant increased risk of death (OR 1.29, 95 per cent c.i. 0.93 to 1.79, P = 0.121). Conclusion: CHOLECOVID provides a unique overview of the treatment of patients with cholecystitis across the globe during the first months of the SARS-CoV-2 pandemic. The study highlights the need for system resilience in retention of elective surgical activity. Cholecystectomy was associated with a low risk of mortality and deferral of treatment results in an increase in avoidable morbidity that represents the non-COVID cost of this pandemic

Vaccination against hepatitis B virus in Spain: a cost-effectiveness analysis

A cost-effectiveness analysis was made to determine the effectiveness of the following strategies of mass immunization with the new recombinant vaccine against the hepatitis B virus in Spain: vaccination of-adolescents, newborns, both populations, and vaccination plus passive immunization of newborns of HBsAg positive mothers. Decision trees supported on Markov models with Monte Carlo simulation have been used for the calculation of costs of the disease, and a mathematical model of differential equations was used for the simulation of the potential effectiveness of vaccination. The costs considered were those associated with the vaccination and travel of subjects, diagnosis, and treatment of the disease. The results are presented as additional cost or saving per case of infection prevented In all assumptions, results showed that the most effective strategy for mass vaccination was the combination of vaccinating all adolescents together with active and passive immunization of children born to HBsAg positive mothers

NOD1 downregulates intestinal serotonin transporter and interacts with other pattern recognition receptors

Serotonin (5-HT) is an essential gastrointestinal modulator whose effects regulate the intestinal physiology. 5-HT effects depend on extracellular 5-HT bioavailability, which is controlled by the serotonin transporter (SERT) expressed in both the apical and basolateral membranes of enterocytes. SERT is a critical target for regulating 5-HT levels and consequently, modulating the intestinal physiology. The deregulation of innate immune receptors has been extensively studied in inflammatory bowel diseases (IBD), where an exacerbated defense response to commensal microbiota is observed. Interestingly, many innate immune receptors seem to affect the serotonergic system, demonstrating a new way in which microbiota could modulate the intestinal physiology. Therefore, our aim was to analyze the effects of NOD1 activation on SERT function, as well as NOD1's interaction with other immune receptors such as TLR2 and TLR4. Our results showed that NOD1 activation inhibits SERT activity and expression in Caco-2/TC7 cells through the extracellular signal-regulated kinase (ERK) signaling pathway. A negative feedback between 5-HT and NOD1 expression was also described. The results showed that TLR2 and TLR4 activation seems to regulate NOD1 expression in Caco-2/TC7 cells. To assess the extend of cross-talk between NOD1 and TLRs, NOD1 expression was measured in the intestinal tract (ileum and colon) of wild type mice and mice with individual knockouts of TLR2, and TLR4 as well as double knockout TLR2/TLR4 mice. Hence, we demonstrate that NOD1 acts on the serotonergic system decreasing SERT activity and molecular expression. Additionally, NOD1 expression seems to be modulated by 5-HT and other immune receptors as TLR2 and TLR4. This study could clarify the relation between both the intestinal serotonergic system and innate immune system, and their implications in intestinal inflammation

Systemic Exosomal Delivery of shRNA Minicircles Prevents Parkinsonian Pathology

The development of new therapies to slow-down or halt Parkinson’s disease progression is a healthcare priority. A key pathological feature is the presence of alpha-synuclein aggregates and there is increasing evidence that alpha-synuclein propagation plays a central role in disease progression. Consequently the down-regulation of alpha-synuclein is a potential therapeutic target. As a chronic disease, the ideal treatment will be minimally invasive and effective in the long-term. Knockdown of gene expression has clear potential and siRNAs specific to alpha-synuclein have been designed, however the efficacy of siRNA treatment is limited by its short-term efficacy. To combat this we designed shRNA minicircles (shRNA-MC), with the potential for prolonged effectiveness, and used RVG-exosomes as vehicle for specific delivery into the brain. We optimized this system using transgenic mice expressing green fluorescent protein (GFP) and demonstrated its ability to down-regulate GFP protein expression in the brain for up to 6 weeks. RVG exosomes were used to deliver anti alpha-synuclein shRNA-MC therapy to the alpha-synuclein preformed fibrils induced model of parkinsonism. This therapy decreased alpha-synuclein aggregation, reduced the loss of dopaminergic neurones and improved the clinical symptoms. Our results confirm the therapeutic potential of shRNA-MC delivered by RVG-exosomes for long-term treatment of neurodegenerative diseases