Metabotropic glutamate receptor blockade reduces preservation damage in livers from donors after cardiac death

We previously demonstrated that the blockade of mGluR5 by 2-methyl-6(phenylethynyl)pyridine (MPEP) reduces both cold and warm ischemia/reperfusion injury. Here we evaluated whether MPEP reduces the hepatic preservation injury in rat livers from cardiac-death-donors (DCDs). Livers from DCD rats were isolated after an in situ warm ischemia (30 min) and preserved for 22 h at 4 °C with UW solution. Next, 10 mg/Kg MPEP or vehicle were administered 30 min before the portal clamping and added to the UW solution (3 µM). LDH released during washout was quantified. Liver samples were collected for iNOS, eNOS, NO, TNF-α, ICAM-1, caspase-3 and caspase-9 protein expression and nuclear factor-erythroid-2-related factor-2 (Nrf2) gene analysis. Lower LDH levels were detected in control grafts versus DCD groups. An increase in eNOS and NO content occurred after MPEP treatment; iNOS and TNF-α content was unchanged. ICAM-1 expression was reduced in the MPEP-treated livers as well as the levels of caspase-3 and caspase-9. Nrf2, oxidative stress-sensitive gene, was recovered to control value by MPEP. These results suggest that MPEP can be used to reclaim DCD livers subjected to an additional period of cold ischemia during hypothermic storage. MPEP protects against apoptosis and increased eNOS, whose overexpression has been previously demonstrated to be protective in hepatic ischemia/reperfusion damage

Transient expression of reck under hepatic ischemia/reperfusion conditions is associated with mapk signaling pathways

In this study, we demonstrated the involvement of matrix metalloproteinases (MMPs) in hepatic ischemia/reperfusion (I/R) injury. Our aim is to evaluate the impact of reperfusion on I/R-related changes in RECK, an MMP modulator, and mitogen-activated protein kinase (MAPKs) pathways (ERK, p38, and JNK). Male Wistar rats were either subjected to 60 min partial-hepatic ischemia or sham-operated. After a 60 min or 120 min reperfusion, liver samples were collected for analysis of MMP-2 and MMP-9 by zymography and RECK, TIMP-1, and TIMP-2 content, MAPKs activation (ERK1/2, JNK1/2, and p38), as well as iNOS and eNOS by Western blot. Serum enzymes AST, ALT, and alkaline-phosphatase were quantified. A transitory decrease in hepatic RECK and TIMPs was associated with a transitory increase in both MMP-2 and MMP-9 activity and a robust activation of ERK1/2, JNK1/2, and p38 were detected at 60 min reperfusion. Hepatic expression of iNOS was maximally upregulated at 120 min reperfusion. An increase in eNOS was detected at 120 min reperfusion. I/R evoked significant hepatic injury in a time-dependent manner. These findings provide new insights into the underlying molecular mechanisms of reperfusion in inducing hepatic injury: a transitory decrease in RECK and TIMPs and increases in both MAPK and MMP activity suggest their role as triggering factors of the organ dysfunction

Effect of chitosan essential oil films on the storage-keeping quality of pork meat products

Edible films based on chitosan were prepared, with and without basil or thyme essential oils, with the aim of assessing their protective ability against lipid oxidation and their antimicrobial activity. Chitosan films had good oxygenbarrier properties, which were worsened by essential oil addition, especially when the film equilibrium moisture content increased. Due to the oxygen-barrier effect, all the films effectively protected pork fat from oxidation, in comparison to unprotected samples. In spite of the worsening of the oxygen-barrier properties, the films with essential oils were more effective than those of pure chitosan, which points to the chemical action of specific antioxidant compounds of the oils. Films were effective to control microbial growth in minced pork meat, although the incorporation of essential oils did not improve their antimicrobial activity. Throughout the storage, the films led to colour changes in minced pork meat associated with the conversion of myoglobin into metmyoglobin due to the reduction of the oxygen availability.The authors acknowledge the financial support provided by the Universitat Politecnica de Valencia (PAID-06-09-2834), Generalitat Valenciana (GV/2010/082) and Ministerio de Educacion y Ciencia (AGL2010-20694). Author J. Bonilla is deeply grateful to Generalitat Valenciana for a Santiago Grisolia Grant.Bonilla Lagos, MJ.; Vargas, M.; Atarés Huerta, LM.; Chiralt Boix, MA. (2014). Effect of chitosan essential oil films on the storage-keeping quality of pork meat products. Food and Bioprocess Technology. 7(8):2443-2450. https://doi.org/10.1007/s11947-014-1329-3S2443245078ASTM D3985. (1995). Standard test method for oxygen gas transmission rate through plastic films and sheeting using a coulometric sensor. West Conshohocken: American Society for Testing and Materials.Atarés, L., Pérez-Masiá, R., & Chiralt, A. (2011). The role of some antioxidants in the HPMC film properties and lipid protection in coated toasted almonds. Journal of Food Engineering, 104, 649–656.Aureli, P., Costantini, A., & Zolea, S. (1992). Antimicrobial activity of some plant essential oils against Listeria monocytogenes. Journal of Food Protection, 55, 344–348.Baranauskiene, R., Venskutoni, S. P. R., Viskelis, P., & Dambrauskiene, E. (2003). Influence of nitrogen fertilizers on the yield and composition of thyme (Thymus vulgaris). Journal of Agricultural and Food Chemistry, 51, 7751–7758.Bonilla, J., Atarés, L., Vargas, M., & Chiralt, A. (2012a). Edible films and coatings to prevent the detrimental effect of oxygen on food quality: possibilities and limitations. Journal of Food Engineering, 110, 208–213.Bonilla, J., Atarés, L., Vargas, M., & Chiralt, A. (2012b). Effect of essential oils and homogenization conditions on properties of chitosan-based films. Food Hydrocolloids, 26, 9–16.Burt, S. (2004). Essential oils: their antibacterial properties and potential applications in foods—a review. International Journal of Food Microbiology, 94, 223–253.Burt, S. A., & Reinders, R. D. (2003). Antibacterial activity of selected plant essential oils against Escherichia coli O157:H7. Letters in Applied Microbiology, 36, 162–167.Caner, C., Vergano, P. J., & Wiles, J. L. (1998). Chitosan film mechanical and permeation properties as affected by acid, plasticizer and storage. Journal of Food Science, 63, 1049–1053.Casariego, A., Souza, B. W. S., Cerqueira, M. A., Teixeira, J. A., Cruz, L., Díaz, R., et al. (2009). Chitosan/clay ‘films properties as affected by biopolymer and clay micro/nanoparticles’ concentrations. Food Hydrocolloids, 23, 1895–1902.Devlieghere, F., Vermeiren, L., & Debevere, J. (2004). New preservation technologies: possibilities and limitations. International Dairy Journal, 14, 273–285.Di Pasqua, R., Hoskins, N., Betts, G., & Mauriello, G. (2006). Changes in membrane fatty acids composition of microbial cells induced by addiction of thymol, carvacrol, limonene, cinnamaldehyde and eugenol in the growing media. Journal of Agricultural and Food Chemistry, 54, 2745–2749.Di Pierro, P., Sorrentino, A., Mariniello, L., Giosafatto, C. V. L., & Porta, R. (2011). Chitosan/whey protein film as active coating to extend Ricotta cheese shelf-life. LWT--Food Science and Technology, 44, 2324–2327.Fabra, M. J., Talens, P., Gavara, R., & Chiralt, A. (2012). Barrier properties of sodium caseinate films as affected by lipid composition and moisture content. Journal of Food Engineering, 109(3), 372–379.Gaysinsky, S., Davidson, P. M., Bruce, B. D., & Weiss, J. (2005). Growth inhibition of E. Coli O157:H7 and Listeria monocytogenes by carvacrol and eugenol encapsulated in surfactant micelles. Journal of Food Protection, 68, 2559–2566.Govaris, A., Botsoglou, E., Sergelidis, D., & Chatzopoulou, P. D. (2011). Antibacterial activity of oregano and thyme essential oils against Listeria monocytogenes and Escherichia coli O157:H7 in feta cheese packaged under modified atmosphere. LWT - Food Science and Technology, 44, 1240–1244.Han, J. H., & Gennadios, A. (2005). Edible films and coatings: a review. In J. H. Han (Ed.), Innovations in Food Packaging (pp. 39–262). Oxford: Elsevier Academic.Kim, J., Marshall, M. R., & Wei, C. I. (1995). Antibacterial activity of some essential oil components against five foodborne pathogens. Journal of Agricultural and Food Chemistry, 43, 2839–2845.Labuza, T. P. (1980). The effect of water activity on reaction kinetics of food deterioration. Food Technology, 34, 36–41.Mancini, R. A., & Hunt, M. C. (2005). Current research in meat color. Meat Science, 71, 100–121.Moure, A., Cruz, J. M., Franco, D., Dominguez, J. M., Sineiro, J., Dominguez, H., et al. (2001). Natural antioxidants from residual sources. Food Chemistry, 72, 145–171.Rao, M. S., Chander, R., & Sharma, A. (2005). Development of shelf-stable intermediate moisture meat products using active edible chitosan coating and irradiation. Journal of Food Science, 70, 325–331.Salame, M. (1986). Barrier polymers. In M. Bakker (Ed.), The Wiley encyclopedia of packaging technology (pp. 48–54). New York: Wiley.Sánchez-González, L., González-Martínez, C., Chiralt, A., & Cháfer, M. (2010). Physical and antimicrobial properties of chitosan–tea tree essential oil composite films. Journal of Food Engineering, 98, 443–452.Sánchez-González, L., Vargas, M., González-Martínez, C., Chiralt, A., & Cháfer, M. (2011a). Use of essential oils in bioactive edible coatings. Food Engineering Reviews, 3, 1–16.Sánchez-González, L., Cháfer, M., Hernández, M., Chiralt, A., & González-Martínez, C. (2011b). Antimicrobial activity of polysaccharide films containing essential oils. Food Control, 22, 1302–1310.Seydim, A. C., & Sarikus, G. (2006). Antimicrobial activity of whey protein based edible films incorporated with oregano, rosemary and garlic essential oils. Food Research International, 39, 639–644.Shan, B., Cai, Y. Z., Sun, M., & Corke, H. (2005). Antioxidant capacity of 26 spice extracts and characterization of their phenolic constituents. Journal of Agricultural and Food Chemistry, 53, 7749–7759.Singh, B., Falahee, M. B., & Adams, M. R. (2001). Synergistic inhibition of Listeria monocytogenes by nisin and garlic extract. Food Microbioliology, 18, 133–139.Vargas, M., Albors, A., Chiralt, A., & González-Martínez, C. (2006). Quality of cold-stored strawberries as affected by chitosan–oleic acid edible coatings. Postharvest Biology and Technology, 41, 164–171.Vargas, M., Albors, A., Chiralt, A., & González-Martínez, C. (2009). Characterization of chitosan–oleic acid composite films. Food Hydrocolloids, 23, 536–547.Vargas, M., Albors, A., & Chiralt, A. (2011). Application of chitosan-sunflower oil edible films to pork meat hamburgers. Procedia Food Science, 1, 39–43.Wan, J., Wilcock, A., & Coventry, M. J. (1998). The effect of essential oils of basil on the growth of Aeromonas hydrophila and Pseudomonas fluorescens. Journal of Applied Microbiology, 84, 152–158.Zivanovic, S., Chi, S., & Draughon, F. (2005). Antimicrobial activity of chitosan films enriched with essential oils. Journal of Food Science, 70, 45–51

Performances in cerebellar and neuromuscular transmission tests are correlated in migraine with aura

In previous studies, we described subclinical abnormalities of neuromuscular transmission and cerebellar functions in migraineurs. The aim of this study was to search if these two functions are correlated in the same patient. Thirteen migraineurs [five without aura (MO) and eight with aura (MA)] underwent both stimulation-SFEMG and 3D-movement analysis. Single fiber EMG (SFEMG) results were expressed as the “mean value of consecutive differences” (mean MCD). Precision of arm-reaching movements (measured with an infrared optoelectronic tracking system) was expressed as the average deviation in the horizontal plane. Median values of mean MCD and mean horizontal deviation were not different between MO and MA. However, in MA, but not in MO, both variables were positively correlated. Thus, we conclude that neuromuscular transmission and cerebellar functions are correlated in the same patient when affected by migraine with aura. We suggest that this correlation might be due to a common molecular abnormality

In Vitro Downregulation of Matrix Metalloproteinase-9 in Rat Glial Cells by CCR5 Antagonist Maraviroc: Therapeutic Implication for HIV Brain Infection

BACKGROUND: Matrix metalloproteinases (MMPs) released by glial cells are important mediators of neuroinflammation and neurologic damage in HIV infection. The use of antiretroviral drugs able to combat the detrimental effect of chronic inflammation and target the exaggerated MMP activity might represent an attractive therapeutic challenge. Recent studies suggest that CCR5 antagonist maraviroc (MVC) exerts immunomodulant and anti-inflammatory activity beyond its anti-HIV properties. We investigated the in vitro effect of MVC on the activity of MMPs in astrocyte and microglia cultures. METHODOLOGY/PRINCIPAL FINDINGS: Primary cultures of rat astrocytes and microglia were activated by exposure to phorbol myristate acetate (PMA) or lypopolysaccharide (LPS) and treated in vitro with MVC. Culture supernatants were subjected to gelatin zymography and quantitative determination of MMP-9 and MMP-2 was done by computerized scanning densitometry. MMP-9 levels were significantly elevated in culture supernatants from both LPS- and PMA-activated astrocytes and microglia in comparison to controls. The treatment with MVC significantly inhibited in a dose-dependent manner the levels and expression of MMP-9 in PMA-activated astrocytes (p<0,05) and, to a lesser extent, in PMA-activated microglia. By contrast, levels of MMP-2 did not significantly change, although a tendency to decrease was seen in PMA-activated astrocytes after treatment with MVC. The inhibition of levels and expression of MMP-9 in PMA-activated glial cells did not depend on cytotoxic effects of MVC. No inhibition of MMP-9 and MMP-2 were found in both LPS-activated astrocytes and microglia. CONCLUSIONS: The present in vitro study suggests that CCR5 antagonist compounds, through their ability to inhibit MMP-9 expression and levels, might have a great potential for the treatment of HIV-associated neurologic damage

Gabapentin as add-on to morphine for severe neuropathic or mixed pain in children from age 3 months to 18 years - Evaluation of the safety, pharmacokinetics, and efficacy of a new gabapentin liquid formulation: Study protocol for a randomized controlled trial

Background: Gabapentin has shown efficacy in the treatment of chronic neuropathic or mixed pain in adults. Although pediatric pain specialists have extensive experience with gabapentin for the treatment of neuropathic pain, its use is off-label. Its efficacy and safety in this context have never been shown. The aim of this trial is to compare gabapentin with placebo as add-on to morphine for the treatment of severe chronic mixed or neuropathic pain in children. This trial is part of the European Union Seventh Framework Programme project Gabapentin in Paediatric Pain (GAPP) to develop a pediatric use marketing authorization for a new gabapentin suspension. Methods/design: The GAPP-2 study is a randomized, double-blind, placebo-controlled, multicenter superiority phase II study in children with severe chronic neuropathic or mixed pain. Its primary objective is to evaluate the efficacy of a gabapentin liquid formulation as adjunctive therapy to morphine. Sixty-six eligible children 3 months to 18 years of age with severe pain (pain scores ≥ 7), stratified in three age groups, will be randomized to receive gabapentin (to an accumulating dose of 45 to 63 mg/kg/day, dependent on age) or placebo, both in addition to morphine, for 12 weeks. Randomization will be preceded by a short washout period, and treatment will be initiated by a titration period of 3 weeks. After the treatment period, medication will be tapered during 4 weeks. The primary endpoint is the average pain scores in the two treatment groups (average of two measures each day for 3 days before the end-of-study visit [V10] assessed by age-appropriate pain scales (Face, Legs, Activity, Cry, Consolability scale; Faces Pain Scale-Revised; Numeric Rating Scale). Secondary outcomes include percentage responders to treatment (subjects with 30% reduction in pain scale), number of episodes of breakthrough pain, number of rescue interventions, number of pain-free days, participant dropouts, quality of life (Pediatric Quality of Life Inventory), and acceptability of treatment. Outcomes will be measured at the end-of-study visit after 12 weeks of treatment at the optimal gabapentin dose. Groups will be compared on an intention-to-treat basis. Discussion: We hope to provide evidence that the combination of morphine and gabapentin will provide better analgesia than morphine alone and will be safe. We also aim to obtain confirmation of the recommended pediatric dose. Trial registration: EudractCT, 2014-004897-40. Registered on 7 September 2017. ClinicalTrials.gov, NCT03275012. Registered on 7 September 2017