Requirement for sphingosine kinase 1 in mediating phase 1 of the hypotensive response to anandamide in the anaesthetised mouse

In the isolated rat carotid artery, the endocannabinoid anandamide induces endothelium-dependent relaxation via activation of the enzyme sphingosine kinase (SK). This generates sphingosine-1-phosphate (S1P) which can be released from the cell and activates S1P receptors on the endothelium. In anaesthetised mice, anandamide has a well-characterised triphasic effect on blood pressure but the contribution of SK and S1P receptors in mediating changes in blood pressure has never been studied. Therefore, we assessed this in the current study. The peak hypotensive response to 1 and 10 mg/kg anandamide was measured in control C57BL/6 mice and in mice pretreated with selective inhibitors of SK1 (BML-258, also known as SK1-I) or SK2 ((R)-FTY720 methylether (ROMe), a dual SK1/2 inhibitor (SKi) or an S1P1 receptor antagonist (W146). Vasodilator responses to S1P were also studied in isolated mouse aortic rings. The hypotensive response to anandamide was significantly attenuated by BML-258 but not by ROMe. Antagonising S1P1 receptors with W146 completely blocked the fall in systolic but not diastolic blood pressure in response to anandamide. S1P induced vasodilation in denuded aortic rings was blocked by W146 but caused no vasodilation in endothelium-intact rings. This study provides evidence that the SK1/S1P regulatory-axis is necessary for the rapid hypotension induced by anandamide. Generation of S1P in response to anandamide likely activates S1P1 to reduce total peripheral resistance and lower mean arterial pressure. These findings have important implications in our understanding of the hypotensive and cardiovascular actions of cannabinoids

Analysis, assessment, and improvement of fertilizer distribution in pressure irrigation systems

[EN] The transformation of traditional irrigation systems into pressure irrigation networks allows water users associations to use central fertigation systems. For efficient fertigation management, however, it is essential to obtain uniform distribution of the injected fertilizer through the system and to understand the hydraulic processes that take place in the central fertigation system. This will allow users to implement strategies that improve fertilizer distribution. In this work we develop a new methodology to improve fertilizer distribution uniformity and apply it to a case study. The results show how fertilizer distribution can be improved by means of proper scheduling of irrigation deliveries. The best results are obtained when fertigating sectors operate without non-fertigating sectors and there are not intermediate irrigations without fertilizer, achieving an improvement of the fertilizer distribution of 10.5%. In addition, this work highlights the difficulties of obtaining uniform distribution of fertilizer in a centralized irrigation system when there are users that do not want to make use of it.Jiménez Bello, MA.; Martínez Alzamora, F.; Bou Soler, V.; Bartolín Ayala, HJ. (2010). Analysis, assessment, and improvement of fertilizer distribution in pressure irrigation systems. Irrigation Science. 29(1):45-53. doi:10.1007/s00271-010-0215-7S4553291Arviza J y Balbastre I (2002a) “Redes de Riego a presión. Consideraciones sobre la fertirrigación colectiva”. Revista Levante Agrícola nº 359. 1º trimestre. Págs. 70–81. Ediciones y promociones LAV SL. ValenciaArviza J y Balbastre I (2002b) “Redes de Riego a presión. Consideraciones sobre la fertirrigación colectiva. Parte II”. Revista Levante Agrícola nº 360. 2º trimestre. Págs. 133–139. Ediciones y promociones LAV SL. ValenciaArviza J, Martínez F, y Balbastre I (2002) Análisis de la distribución de fertilizantes en sistemas colectivos de riego a presión. XX Congreso Nacional de Riegos. Ciudad Real. EspañaBracy RP, Parish RL, Rosendale RM (2003) Fertigation uniformity affected by injector type. Horttechnology 13:103–105Goldberg DE (1989) Genetic algorithms in search, optimization and machine learning. Addison-Wesley, ReadingJiménez MA, Martínez F, Arviza J, Manzano J (2006) Herramientas para el uso racional del agua con el apoyo de un GIS (HuraGIS). XXIV. Congreso nacional de riegos. Lugo (Spain). (Jun 2006)Jiménez MA, Martínez F, Arviza J, Manzano J (2008) Optimización de la sectorización de redes de riego a presión mediante algoritmos genéticos XXVI. Congreso nacional de riegos. Huesca. (Jun 2008)Jiusheng L, Yibin M, Bei L (2007) Field evaluation of fertigation uniformity as affected by injector type and manufacturing variability of emitters. Irrigation Sci 25:117–125Kalyanmoy D (2001) Multi-objective optimization using evolutionary algorithms. Willey, EnglandRossman LA (2000) Epanet 2, Users Manual. Water Supply and Water Resources Division. National Risk Management Research Laboratory, CincinnatiSavic D, Walters G (1997) Genetic algorithms for least-cost design of water distribution networks. J Water Resour Plann Manag 123(2):67–77 (March/April 1997

Hyperinsulinemic-euglycemic Clamps in Conscious, Unrestrained Mice

Type 2 diabetes is characterized by a defect in insulin action. The hyperinsulinemic-euglycemic clamp, or insulin clamp, is widely considered the "gold standard" method for assessing insulin action in vivo. During an insulin clamp, hyperinsulinemia is achieved by a constant insulin infusion. Euglycemia is maintained via a concomitant glucose infusion at a variable rate. This variable glucose infusion rate (GIR) is determined by measuring blood glucose at brief intervals throughout the experiment and adjusting the GIR accordingly. The GIR is indicative of whole-body insulin action, as mice with enhanced insulin action require a greater GIR. The insulin clamp can incorporate administration of isotopic 2[14C]deoxyglucose to assess tissue-specific glucose uptake and [3-3H]glucose to assess the ability of insulin to suppress the rate of endogenous glucose appearance (endoRa), a marker of hepatic glucose production, and to stimulate the rate of whole-body glucose disappearance (Rd)

Simultaneous bilateral hip replacement reveals superior outcome and fewer complications than two-stage procedures: a prospective study including 1819 patients and 5801 follow-ups from a total joint replacement registry

<p>Abstract</p> <p>Background</p> <p>Total joint replacements represent a considerable part of day-to-day orthopaedic routine and a substantial proportion of patients undergoing unilateral total hip arthroplasty require a contralateral treatment after the first operation. This report compares complications and functional outcome of simultaneous versus early and delayed two-stage bilateral THA over a five-year follow-up period.</p> <p>Methods</p> <p>The study is a post hoc analysis of prospectively collected data in the framework of the European IDES hip registry. The database query resulted in 1819 patients with 5801 follow-ups treated with bilateral THA between 1965 and 2002. According to the timing of the two operations the sample was divided into three groups: I) 247 patients with simultaneous bilateral THA, II) 737 patients with two-stage bilateral THA within six months, III) 835 patients with two-stage bilateral THA between six months and five years.</p> <p>Results</p> <p>Whereas postoperative hip pain and flexion did not differ between the groups, the best walking capacity was observed in group I and the worst in group III. The rate of intraoperative complications in the first group was comparable to that of the second. The frequency of postoperative local and systemic complication in group I was the lowest of the three groups. The highest rate of complications was observed in group III.</p> <p>Conclusions</p> <p>From the point of view of possible intra- and postoperative complications, one-stage bilateral THA is equally safe or safer than two-stage interventions. Additionally, from an outcome perspective the one-stage procedure can be considered to be advantageous.</p

Small-molecule allosteric activators of PDE4 long form cyclic AMP phosphodiesterases

Cyclic AMP (cAMP) phosphodiesterase-4 (PDE4) enzymes degrade cAMP and underpin the compartmentalization of cAMP signaling through their targeting to particular protein complexes and intracellular locales. We describe the discovery and characterization of a small-molecule compound that allosterically activates PDE4 long isoforms. This PDE4-specific activator displays reversible, noncompetitive kinetics of activation (increased Vmax with unchanged Km), phenocopies the ability of protein kinase A (PKA) to activate PDE4 long isoforms endogenously, and requires a dimeric enzyme assembly, as adopted by long, but not by short (monomeric), PDE4 isoforms. Abnormally elevated levels of cAMP provide a critical driver of the underpinning molecular pathology of autosomal dominant polycystic kidney disease (ADPKD) by promoting cyst formation that, ultimately, culminates in renal failure. Using both animal and human cell models of ADPKD, including ADPKD patient-derived primary cell cultures, we demonstrate that treatment with the prototypical PDE4 activator compound lowers intracellular cAMP levels, restrains cAMP-mediated signaling events, and profoundly inhibits cyst formation. PDE4 activator compounds thus have potential as therapeutics for treating disease driven by elevated cAMP signaling as well as providing a tool for evaluating the action of long PDE4 isoforms in regulating cAMP-mediated cellular processes