Toll-like receptor 9 modifies intestinal serotonergic system.

Introduction: Toll-like receptor 9 (TLR9) is expressed in intestinal epithelial cells, which recognize microbiota developing different responses 1. Several studies have shown that TLR9 seems to be involved in Inflammatory Bowel Diseases (IBD) due to an inappropriate defensive response against microorganisms 2. Moreover, intestinal serotonergic system is also altered in IBD, where extracellular serotonin (5–HT) levels are increased 3. 5-HT bioavailability is mainly regulated by the serotonin transporter (SERT), expressed in enterocytes 4. Aims & Methods: The aim of the present study was to analyse whether TLR9 activation affects SERT expression and activity, and expression of other elements from the serotonergic system (TPH1, TPH2 and 5-HT receptors). Human enterocyte-like Caco-2 cells, and ileum and colon from TLR9-/- mice and Dextran Sulphate Sodium (DSS) mouse colitis model were used as experimental models. mRNA expression was determined by RT-qPCR, and protein expression by western blot..

Toll-like receptor 9 activation affects intestinal serotonin transporter activity and expression in Caco-2 cells

Background: Toll-like Receptor 9 (TLR9) is expressed mainly in the endosomal membrane of intestinal cells and mediates intestinal host-microbiota interaction. Serotonin (5-HT) is an intestinal neuromodulator involved in the intestinal immunity and homeostasis. In addition, a high level of 5-HT has been described in intestinal inflammation. 5-HT intestinal availability is mainly regulated by the serotonin transporter (SERT) expressed in enterocytes. Aim: The interaction of TLR9 with serotoninergic system remains known. Therefore, the aim of the present study was to assess the effects of TLR9 activation on SERT activity and expression. Methods: Caco-2 cells and colon from wild type (WT) and TLR9 C57BL/10 mice were used in this study. SERT activity (5-HT uptake) in Caco-2 cells and SERT expression (RT-qPCR and western blotting) in both Caco-2 cells and colon from WT and TLR9 mice, were analyzed. TLR9 mRNA and protein levels were also measured in Caco-2 cells. Results: TLR9 activation in Caco-2 cells reduced SERT activity in a MyD88 independent-way. SERT mRNA and protein level in both cell lysate and brush border membrane, were also diminished. SERT protein expression in colon of TLR9 mice resulted augmented compared with WT mice. Interestingly, activation of TLR9 in Caco-2 cells diminished TLR9 mRNA and protein in the cell lysate; however, TLR9 protein in brush border resulted increased. Conclusions: The results of this work highlight the role of TLR9 as a mediator intestinal homeostasis and/or intestinal inflammation by regulating intestinal serotoninergic system

3D VMAT Verification Based on Monte Carlo Log File Simulation with Experimental Feedback from Film Dosimetry.

A model based on a specific phantom, called QuAArC, has been designed for the evaluation of planning and verification systems of complex radiotherapy treatments, such as volumetric modulated arc therapy (VMAT). This model uses the high accuracy provided by the Monte Carlo (MC) simulation of log files and allows the experimental feedback from the high spatial resolution of films hosted in QuAArC. This cylindrical phantom was specifically designed to host films rolled at different radial distances able to take into account the entrance fluence and the 3D dose distribution. Ionization chamber measurements are also included in the feedback process for absolute dose considerations. In this way, automated MC simulation of treatment log files is implemented to calculate the actual delivery geometries, while the monitor units are experimentally adjusted to reconstruct the dose-volume histogram (DVH) on the patient CT. Prostate and head and neck clinical cases, previously planned with Monaco and Pinnacle treatment planning systems and verified with two different commercial systems (Delta4 and COMPASS), were selected in order to test operational feasibility of the proposed model. The proper operation of the feedback procedure was proved through the achieved high agreement between reconstructed dose distributions and the film measure- ments (global gamma passing rates > 90% for the 2%/2 mm criteria). The necessary discre- tization level of the log file for dose calculation and the potential mismatching between calculated control points and detection grid in the verification process were discussed. Besides the effect of dose calculation accuracy of the analytic algorithm implemented in treatment planning systems for a dynamic technique, it was discussed the importance of the detection density level and its location in VMAT specific phantom to obtain a more reliable DVH in the patient CT. The proposed model also showed enough robustness and efficiency to be considered as a pre-treatment VMAT verification system.Ministerio de Ciencia y Tecnología SAF2011- 27116; IPT-2011-1480-900000

Impact of Using Different Levels of Threshold-Based Artefact Correction on the Quantification of Heart Rate Variability in Three Independent Human Cohorts

The following are available online at https://www.mdpi.com/2077-0383/9/2/325/s1, Figure S1: Example of a visual inspection of a R-R signal to find possible artefacts or premature contractions across Kubios filters; Figures S2 and S3: Differences on the Heart Rate Variability (HRV) time- and frequency-domains parameters respectively without considering the Very Strong filter; Figure S4: Differences between cohorts on the SDNN using different Kubios filters; Figure S5: Differences between cohorts on the pNN50 using different Kubios filters; and Figure S6: Differences between cohorts on the HF using different Kubios filters.Juan M.A. Alcantara, Abel Plaza-Florido, Jairo H. Migueles, Guillermo Sanchez-Delgado and Francisco J. Amaro-Gahete are supported by the Spanish Ministry of Education, Culture and Sport (FPU15/04059, FPU16/02760, FPU15/02645, FPU13/04365 and FPU14/04172 respectively). Guillermo Sanchez-Delgado is supported by the University of Granada Plan Propio de Investigación 2018 (Programa Contratos-Puente and Programa Perfeccionamiento de Docotres). Francisco J. Amaro-Gahete is supported by the University of Granada Plan Propio de Investigación 2019 (Programa Contratos-Puente). Guillermo Sanchez-Delgado and Borja Martinez-Tellez are supported by individual postdoctoral grants from the Fundación Alfonso Martin Escudero.Heart rate variability (HRV) is a non-invasive indicator of autonomic nervous system function. HRV recordings show artefacts due to technical and/or biological issues. The Kubios software is one of the most used software to process HRV recordings, offering different levels of threshold-based artefact correction (i.e., Kubios filters). The aim of the study was to analyze the impact of different Kubios filters on the quantification of HRV derived parameters from short-term recordings in three independent human cohorts. A total of 312 participants were included: 107 children with overweight/obesity (10.0 ± 1.1 years, 58% men), 132 young adults (22.2 ± 2.2 years, 33% men) and 73 middle-aged adults (53.6 ± 5.2 years, 48% men). HRV was assessed using a heart rate monitor during 10–15 min, and the Kubios software was used for HRV data processing using all the Kubios filters available (i.e., 6). Repeated-measures analysis of variance indicated significant differences in HRV derived parameters in the time-domain (all p < 0.001) across the Kubios filters in all cohorts, moreover similar results were observed in the frequency-domain. When comparing two extreme Kubios filters, these statistical differences could be clinically relevant, e.g. more than 10 ms in the standard deviation of all normal R-R intervals (SDNN). In conclusion, the results of the present study suggest that the application of different Kubios filters had a significant impact on HRV derived parameters obtained from short-term recordings in both time and frequency-domains.The study was funded by the Spanish Ministry of Economy and Competitiveness (DEP2013-47540 and DEP2016-79512-R), the Fondo de Investigación Sanitaria del Instituto de Salud Carlos III (PI13/01393), European Union Development Funds, the Fundación Iberoamericana de Nutrición (FINUT), the Redes Temáticas de Investigación Cooperativa RETIC (Red SAMID RD16/0022), the University of Granada Plan Propio de Investigación 2016 (Excellence actions: Unit of Excellence on Exercise and Health [UCEES]), and the Junta de Andalucía, Consejería de Conocimiento, Investigación y Universidades (FEDER: ref. SOMM17/6107/UGR)

Estado de estrés oxidativo en el hígado graso experimental en ratones machos y su proyección sobre la funcionalidad hepática

En medicina veterinaria, el rol del estrés oxidativo (EO) en la producción y reproducción animal ha adquirido relevancia debido al deterioro de ambas funciones en animales con hepatoesteatosis o hígado graso (HG). Este estudio tuvo el objetivo de determinar si en el HG experimental causado por etionina en ratones machos NMRI adultos, se establecía un estado de EO hepático y se alteraba la función del hígado. Se utilizaron dos grupos de 10 animales: uno control y otro tratado con DL-etionina por vía intraperitoneal en dosis de 7,5 mg/20 g de peso corporal. La magnitud y características del HG fueron determinadas histológicamente y la cuantía del depósito graso hepático se estableció por la concentración de triglicéridos, análisis que corroboraron la generación de hepatoesteatosis en los machos inyectados con DL-etionina. Los productos de la degradación lipoperoxidativa, dienos conjugados (DC) y malondialdehído (MDA), indicadores de EO celular, fueron cuantificados por espectrofotometría, mediante su concentración en el homogeneizado hepático. El daño en la funcionalidad hepática se cuantificó por los niveles plasmáticos de la alanina aminotransferasa (ALT) y de la aspartato aminotransferasa (AST), mediante kits comerciales. La inducción de HG causó una elevación significativa de los DC: de 231,18 ± 15,53 µmoles/mg proteínas a 297,45 ± 23,10 mmoles/mg proteínas (P<0,05), así como del MDA: de 364,91 ± 17,73 nmoles/mg proteínas a 852,91 ± 55,26 nmoles/mg proteínas (P<0,001). En los ratones con HG, la actividad plasmática de las aminotransferasas aumentó significativamente: ALT de 59,40 ± 5,16 U/l a 169,86 ± 18,78 U/l (P<0,001) y AST de 158,35 ± 13,54 U/l a 241,93 ± 10,14 U/l (P<0,05). Estos resultados muestran que en el HG inducido por etionina en ratones machos NMRI se produce un estado de EO que podría ser responsable de la alteración en la funcionalidad hepática. In veterinary medicine the role of oxidative stress (OS) in animal production and reproduction has become important due to the alteration of both functions in animals with hepatosteatosis or fatty liver (FL). This study aimed to determine whether a state of hepatic OS was established in the experimental FL caused by ethionine in adult male mice NMRI, and whether liver function was altered. Two groups of 10 animals were used: one control and another treated with DL-ethionine intraperitoneally at 7.5 mg/20 g body weight. The magnitude and characteristics of FL were determined histologically and the amount of liver fat depot was established by the concentration of triglycerides. Both analysis corroborated generation of hepatosteatosis in males injected with DL-ethionine. Products from lipoperoxidative degradation (conjugated dienes (CD) and malondialdehyde (MDA), indicators of cellular OS, were quantified spectrophotometrically by its concentration in liver homogenate. Damage to liver function was measured by plasmatic levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST), using commercial kits. The induction of FL caused a significant rise in CD from 231.18 ± 15.53 µmoles/mg protein to 297.45 ± 23.10 mmoles/mg protein (P<0.05), as well as the MDA: from 364.91 ± 17.73 nmoles/mg protein to 852.91 ± 55.26 nmoles/mg protein (P<0.001). In mice with FL aminotransferases plasmatic activity increased significantly: ALT of 59.40 ± 5.16 U/l from 169.86 ± 18.78 U/l (P<0.001) and AST from 158.35 ± 13.54 U/l to 241.93 ± 10.14 U/l (P<0.05). These results show that in ethionine-induced FL in male NMRI mice a state of OS is induced that could be responsible for the alteration in liver function

LDL particle size and composition and incident cardiovascular disease in a South-European population: The Hortega-Liposcale Follow-up Study.

The association of low-density lipoprotein (LDL) particle composition with cardiovascular risk has not been explored before. The aim was to evaluate the relationship between baseline LDL particle size and composition (proportions of large, medium and small LDL particles over their sum expressed as small-LDL %, medium-LDL % and large-LDL %) and incident cardiovascular disease in a population-based study. Methods: Direct measurement of LDL particles was performed using a two-dimensional NMR-technique (Liposcale®). LDL cholesterol was assessed using both standard photometrical methods and the Liposcale® technique in a representative sample of 1162 adult men and women from Spain. Results: The geometric mean of total LDL particle concentration in the study sample was 827.2 mg/dL (95% CI 814.7, 839.8). During a mean follow-up of 12.4 ± 3.3 years, a total of 159 events occurred. Medium LDL particles were positively associated with all cardiovascular disease, coronary heart disease (CHD) and stroke after adjustment for traditional risk factors and treatment. Regarding LDL particle composition, the multivariable adjusted hazard ratios for CHD for a 5% increase in medium and small LDL % by a corresponding decrease of large LDL % were 1.93 (1.55, 2.39) and 1.41 (1.14, 1.74), respectively. Conclusions: Medium LDL particles were associated with incident cardiovascular disease. LDL particles showed the strongest association with cardiovascular events when the particle composition, rather than the total concentration, was investigated. A change in baseline composition of LDL particles from large to medium and small LDL particles was associated with an increased cardiovascular risk, especially for CHD

ERP and four dimensions of absorptive capacity: lessons from a developing country

Enterprise resource planning systems can grant crucial strategic, operational and information-based benefits to adopting firms when implemented successfully. However, a failed implementation can often result in financial losses rather than profits. Until now, the research on the failures and successes were focused on implementations in large manufacturing and service organizations firms located in western countries, particularly in USA. Nevertheless, IT has gained intense diffusion to developing countries through declining hardware costs and increasing benefits that merits attention as much as developed countries. The aim of this study is to examine the implications of knowledge transfer in a developing country, Turkey, as a paradigm in the knowledge society with a focus on the implementation activities that foster successful installations. We suggest that absorptive capacity is an important characteristic of a firm that explains the success level of such a knowledge transfer.Publicad