Dual-specificity phosphatase 6 regulates CD4+ T-cell functions and restrains spontaneous colitis in IL-10-deficient mice.

Mitogen-activated protein kinase (MAPK) phosphatases are dual-specificity phosphatases (DUSPs) that dephosphorylate phosphothreonine and phosphotyrosine residues within MAPKs. DUSP6 preferentially dephosphorylates extracellular signal-regulated kinases 1 and 2 (ERK1/2) rendering them inactive. Here, we study the role of DUSP6 in CD4(+) T-cell function, differentiation, and inflammatory profile in the colon. Upon T-cell receptor (TCR) stimulation, DUSP6 knockout (Dusp6(-/-)) CD4(+) T cells showed increased ERK1/2 activation, proliferation, T helper 1 differentiation, and interferon-γ production, as well as a marked decrease in survival, interleukin- 17A (IL-17A) secretion, and regulatory T-cell function. To analyze the role of DUSP6 in vivo, we employed the Il10(-/-) model of colitis and generated Il10(-/-)/Dusp6(-/-) double-knockout mice. Il10(-/-)/Dusp6(-/-) mice suffered from accelerated and exacerbated spontaneous colitis, which was prevented by ERK1/2 inhibition. ERK1/2 inhibition also augmented regulatory T-cell differentiation in vitro and in vivo in both C57Bl/6 and Dusp6(-/-) mice. In summary, DUSP6 regulates CD4(+) T-cell activation and differentiation by inhibiting the TCR-dependent ERK1/2 activation. DUSP6 might therefore be a potential intervention target for limiting aberrant T-cell responses in T-cell-mediated diseases, such as inflammatory bowel disease.

Impacts of irrigation network sectoring as an energy saving measure on olive grove production.

In recent years, improving water use efficiency has been one of the most important challenges for the agricultural sector. However, such improvements have led to the installation of pressurized irrigation systems which generally require more energy to operate, especially in plantations on sloping and mountainous lands. Thus, the reduction of energy use in these systems has also become a major issue. Irrigation network sectoring has been proposed as one of the most effective energy saving measures. Typically, however, the potential benefits of this management strategy have been evaluated by means of theoretical approaches in networks that were originally designed to supply water on demand and not after water application in real irrigation districts designed following sectoring strategies. In this work, this measure is applied to an irrigation district devoted to olive grove production in a mountainous area that was designed according to this management strategy. With this aim, the WEBSO (Water and Energy Based Sectoring Operation) algorithm, which was developed in a previous work, has been modified in order to take into account the specific characteristics of the irrigation district and its actual management, as well as to analyze sensitivity to several irrigation water depths in terms of both energy demand and yields. An economic analysis of the potential benefits of this management strategy is also carried out. The results show that this measure has lead to a nearly 30% reduction in energy consumption, while increasing farmers' profits by 13% compared to traditional on-demand operations.

[Bacterial DNA in patients with cirrhosis and sterile ascites. Its role as a marker of bacterial translocation and prognostic tool]. / ADN bacterano en pacientes con cirrosis y ascitis estéril. Papel como marcador de translocación bacteriana y herramienta pronóstica.

During the last decade, we have witnessed an increase in the amount of data related with the presence of bacterial translocation in experimental models of cirrhosis. However, clinical studies have been limited by the lack of non-invasive methods to study this phenomenon. Over the past years, the research developed in our laboratory has been focused on the detection of bacterial DNA in serum and ascitic fluid of patients with cirrhosis and sterile ascites, the clinical and immunological implications of such finding. Initially, by means of a polymerase chain reaction (PCR)-based method and automated nucleotide sequencing, we were able to detect and identify the presence of fragments of bacterial DNA in the mentioned patients with culture-negative, non-neutrocytic ascites. Since then, we have accumulated a core of data suggesting that the presence of bacterial DNA may have an important role not only as a marker of bacterial translocation, but also as a short-term prognostic factor. Here, we discuss the past, present and future of this line of investigation.

ADN bacteriano en pacientes con cirrosis y ascitis estéril. Papel como marcador de translocación bacteriana y herramienta pronóstica / Bacterial DNA in patients with cirrhosis and sterile ascites. Its role as a marker of bacterial translocation and prognostic tool

Durante la última década hemos presenciado un aumento de lacantidad de datos relativos a la presencia de translocación bacterianaen los modelos experimentales de cirrosis. Sin embargo, losestudios clínicos se han visto limitados por la falta de métodos noinvasivos para estudiar dicho fenómeno. En los últimos años, lasinvestigaciones realizadas en nuestro laboratorio se han centradoen la detección del ADN bacteriano en el suero y el líquido ascíticode los pacientes con cirrosis y ascitis estéril, y en las implicacionesclínicas que ello conlleva. Al principio, gracias a un métodobasado en la reacción en cadena de la polimerasa (PCR) y el secuenciamientoautomatizado de nucleótidos, pudimos detectar eidentificar la presencia de fragmentos de ADN bacteriano en dichospacientes con ascitis no neutrocítica y con cultivo negativo.Desde entonces hemos acumulado una serie de datos que indicanque la presencia de ADN bacteriano podría desempeñar un papelimportante no sólo como marcador de translocación bacteriana,sino también como factor pronóstico a corto plazo. Expondremosaquí el pasado, el presente y el futuro de esta línea de investigación

During the last decade, we have witnessed an increase in theamount of data related with the presence of bacterial translocationin experimental models of cirrhosis. However, clinical studies havebeen limited by the lack of non-invasive methods to study this phenomenon.Over the past years, the research developed in our laboratoryhas been focused on the detection of bacterial DNA inserum and ascitic fluid of patients with cirrhosis and sterile ascites,the clinical and immunological implications of such finding. Initially,by means of a polymerase chain reaction (PCR)-based methodand automated nucleotide sequencing, we were able to detect andidentify the presence of fragments of bacterial DNA in the mentionedpatients with culture-negative, non-neutrocytic ascites.Since then, we have accumulated a core of data suggesting thatthe presence of bacterial DNA may have an important role notonly as a marker of bacterial translocation, but also as a shorttermprognostic factor. Here, we discuss the past, present and futureof this line of investigation

Translocation of bacterial DNA from Gram-positive microorganisms is associated with a species-specific inflammatory response in serum and ascitic fluid of patients with cirrhosis.

Translocation of bacterial-DNA in patients with cirrhosis and ascites triggers an innate immune response. Identification of characteristics to which this response is sensitive is relevant from a clinical standpoint. The aim of this study has been to determine if the proinflammatory immune response established in vivo in cirrhotic patients with ascites as a consequence of bacterial-DNA translocation is related to the identified bacterial species and their frequency of cytosine-guanosine content in serum and ascitic fluid. Patients with advanced cirrhosis and ascites were included in the study and distributed into groups I and II according to the absence or presence of bacterial-DNA translocation, respectively. Serum and ascitic fluid levels of proinflammatory cytokines after normalization of bacterial-DNA concentration and the activated form of nuclear factor-kappa B in ascitic fluid pellets were measured by enzyme-linked immunosorbent assay techniques. Translocation of bacterial-DNA with higher cytosine-guanosine content induced the highest cytokine response, which was higher than that in patients without bacterial-DNA translocation. The activated form of nuclear factor-kappa B in ascitic fluid pellets of patients with bacterial-DNA translocation was greater in patients with higher bacterial-DNA cytosine-guanosine content, whereas the amount of total nuclear factor-kappa B remained unaltered. Bacterial-DNA translocation induces a marked immune reaction in vivo in patients with advanced cirrhosis and ascites which is related, among other factors, to the bacterial-DNA cytosine-guanosine content. Therefore, the host's immune response to bacterial-DNA translocation constitutes a species-specific phenomenon.

Influence of time and thermocycling on marginal sealing of several dentin adhesive systems.

This study evaluated the in vitro microleakage of six dentin adhesive systems. Triangle-shaped Class V cavities with coronal margin in enamel and gingival margin in cementum or root dentin were cut in the buccal surfaces of 90 non-carious single-root human teeth. These teeth were randomly assigned into six groups (n = 15) for the evaluation of six different dentin adhesive systems: One Step, Prime & Bond 2.0, Syntac Single, Single Bond, Optibond Solo and Syntac Sprint. The preparations were restored with Degufill Ultra composite and polished using the Enhance system. Each group was randomly divided into three subgroups (n = 5): samples of the first subgroup were immersed in 2% methylene blue solution for seven days; those of the second subgroup remained in a similar solution for 31 days; those of the third subgroup were thermocycled 500x at 5-55 degrees C and immersed in 2% methylene blue for seven days. All 90 teeth were then embedded in methacrylate and bucco-lingually sectioned; the dye penetration was evaluated using an 0-4 ordinal scale. All of the dentin adhesive groups showed minimal leakage at the enamel margins with increased leakage at the gingival margins. Optibond Solo showed the best outcomes among the dentin adhesives tested.

Determination of bisphenol A and related aromatic compounds released from bis-GMA-based composites and sealants by high performance liquid chromatography.

Most of the composites and sealants used in dentistry are based on bisphenol A diglycidylether methacrylate (Bis-GMA). Reports revealed that in situ polymerization is not complete and that free monomers can be detected by different analytic methods. Concerns about the estrogenicity of bisphenol A (BPA) and other aromatic components leached from commercial products have been expressed. We studied biphenolic components eluted from seven composites and one sealant before and after in vitro polymerization using HPLC and gas chromatography/mass spectrometry and we investigated how pH modifications affect the leaching of these components. We found BPA (maximal amount 1.8 microg/mg dental material), its dimethacrylate derivative (Bis-DMA, 1.15 microg/mg), bisphenol A diglycidylether (6. 1 microg/mg), Bis-GMA (2.0 microg/mg), and ethoxylate and propoxylate of bisphenol A in media in which samples of different commercial products were maintained under controlled pH and temperature conditions. Our results confirm the leaching of estrogenic monomers into the environment by Bis-GMA-based composites and sealants in concentrations at which biologic effects have been demonstrated in in vivo experimental models. The main issue with implications for patient care and dentist responsibility is to further determine the clinical relevance of this estrogenic exposure.

Study of the shear bond strength of five one-component adhesives under simulated pulpal pressure.

Recently, several adhesive systems have been introduced that combine the primer and bonding resin in a single bottle. The purpose of this study was to evaluate the bonding efficiency of these one-component adhesives under conditions of simulated pulpal pressure and to determine the influence of storage time on the shear bond strength. One hundred caries-free human molars were embedded with epoxy resin in cylindrical rubber molds. Flat dentin surfaces at a level 1 mm above the pulpal chamber were obtained and used as the region for bonding. The specimens were randomly assigned to five groups (n = 20): (1) Syntac Single, (2) Prime & Bond 2.0, (3) One Step, (4) Single Bond, and (5) OptiBond Solo. Each bonding system was combined with the same composite resin (Herculite XRV). After resin polymerization, half of the samples from each group were tested at 1 week and the other half at 4 weeks. During the bonding procedure and storage time a pulpal pressure of 20 cm of serum was applied. Analysis of the data by one-way ANOVA testing showed that the shear bond strengths were significantly different (P < 0.001). OptiBond Solo and Single Bond presented the best results. As the storage time increased there was a significant decrease in the shear bond strength for all the adhesive systems used.

[Evaluation of in vitro adhesion of cariogenic microorganisms to enamel and dental filling materials]. / Evaluación de la adhesión in vitro de microorganismos cariogénicos en esmalte y materiales de obturación dental.

The adhesive ability of Lactobacillus, Streptococcus mutans and Actinomyces viscosus-naeslundii on enamel, amalgam and composite of microparticle and small-particle are studied "in vitro". The selective mediums used for the three micro-organisms are, respectively, Rogosa agar, M.S.B. and C.F.A.T. The lower adherence is showed by bacterias of Lactobacillus genus. S. mutans and A. viscosus-naeslundii show similar adherence properties between them. The greatest adherence was obtained in composites, being S. mutans the bacteria with a greatest level of adherence to the composites of small-particle, and A. viscosus-naeslundii the bacteria with more adherence to the ones of micro-particle. The adherence on amalgam was slightly lower than the adherence on enamel.