An aPKC-Exocyst Complex Controls Paxillin Phosphorylation and Migration through Localised JNK1 Activation

Atypical protein kinase C (aPKC) isoforms have been implicated in cell polarisation and migration through association with Cdc42 and Par6. In distinct migratory models, the Exocyst complex has been shown to be involved in secretory events and migration. By RNA interference (RNAi) we show that the polarised delivery of the Exocyst to the leading edge of migrating NRK cells is dependent upon aPKCs. Reciprocally we demonstrate that aPKC localisation at the leading edge is dependent upon the Exocyst. The basis of this inter-dependence derives from two-hybrid, mass spectrometry, and co-immunoprecipitation studies, which demonstrate the existence of an aPKC–Exocyst interaction mediated by Kibra. Using RNAi and small molecule inhibitors, the aPKCs, Kibra, and the Exocyst are shown to be required for NRK cell migration and it is further demonstrated that they are necessary for the localized activation of JNK at the leading edge. The migration associated control of JNK by aPKCs determines JNK phosphorylation of the plasma membrane substrate Paxillin, but not the phosphorylation of the nuclear JNK substrate, c-jun. This plasma membrane localized JNK cascade serves to control the stability of focal adhesion complexes, regulating migration. The study integrates the polarising behaviour of aPKCs with the pro-migratory properties of the Exocyst complex, defining a higher order complex associated with the localised activation of JNK at the leading edge of migrating cells that determines migration rate.     

Chemistry matters: biological activity of Eucalyptus essential oils on mosquito larval mortality

The mosquito Aedes aegypti L. (Diptera: Culicidae) is a vector of arboviral diseases such as dengue fever. Currently, the main approach to mosquito control is the application of synthetic insecticides, which can lead to negative environmental impacts and insecticide resistance in mosquito populations. As such, there has been increased interest in developing alternative methods for control of vector populations such as utilizing plant compounds that act as larvicides. The aim of this work is to evaluate the effectiveness of Eucalyptus sp. (Myrtaceae) essential oils for control of Ae. aegypti larvae. The essential oils of seven Eucalyptus species and hybrids were extracted by hydrodistillation and analyzed by gas chromatography coupled to mass spectrometry. The essential oils were further diluted in water with acetone (0.40%) at the following concentrations: 100, 50, 25, and 10 μg ml⁻¹. Mortality trials were conducted in plastic containers with a solution of ultrapure water and 200 μl of diluted oil for a total volume of 50 ml per treatment. The experiments for each Eucalyptus species/hybrid and concentration were performed in triplicate, using a control containing only water and acetone. Twenty larvae were added to each container and mortality was recorded at 1, 2, 4, and 24 h. The Eucalyptus essential oils showed larvicidal activity in most of the evaluated concentrations, mainly at 50 and 100 μg ml⁻¹. Eucalyptus benthamii Maiden & Cambage and the hybrid Urograndis displayed the highest larvicidal potential (100% at 24 h) in the 100 μg ml⁻¹ treatment. Larval mortality of Ae. aegypti showed a positive correlation with the compounds γ-, o-cymol, o-cymene, terpineol, 3-dodecylfuran-2,5-dione, α-pinene, globulol, and ledol. The most abundant compounds identified in the essential oils were 1,8-cineole and α-pinene. These results highlight the potential of using Eucalyptus essential oils for the isolation of natural larvicidal products.     

Efficient in vitro adipocyte model of long-term lipolysis: A tool to study the behavior of lipophilic compounds

The triglycerides (TGs) stored in the white adipose tissue are mobilized during periods of negative energy balance. To date, there is no in vitro model of adipocytes imitating a long period of negative energy balance in which triglycerides are highly mobilized. Such model would allow studying the mobilization of TGs and lipophilic compounds trapped within the adipose tissue (e.g., pollutants and vitamins). The present study aims at developing a performing long-term in vitro lipolysis in adipocytes, resulting in a significant decrease of TG stores. Lipolysis was induced on differentiated rat adipocytes by a lipolytic medium with or without isoproterenol for 12 h. The condition with isoproterenol was duplicated, once with medium renewal every 3 h and once without medium renewal. Adding isoproterenol efficiently triggered lipolysis in a short time (3 h). However, a single stimulation by isoproterenol, without medium renewal, was not sufficient to reduce the TG content during a longer term (12 h). A reesterification of fatty acids occurred after a few hours of lipolysis, resulting in a novel increase of cellular lipids. Regular medium renewal combined with repeated isoproterenol stimulations led to almost emptied cells after 12 h. However, medium renewal without isoproterenol stimulation for 12 h was as efficient in terms of lipid mobilization. Our study demonstrates that, over a short-term period, isoproterenol is required to exert a significant lipolytic effect on adipocytes. During a long-term period, the presence of isoproterenol is no longer essential. Instead, medium renewal becomes the main factor involved in cell emptying. The efficiency of this protocol was demonstrated by visual tracking of the cells and by monitoring the dynamics of release of a lipophilic compound, PCB-153, from adipocytes during lipolysis.     

Dog food production using curcumin as antioxidant: effects of intake on animal growth,health and feed conservation

ABSTRACT

The objectives of this study were to produce dog food containing curcumin replacing synthetic antioxidants, to evaluate its beneficial effects on animal growth and health. Curcumin (100 mg/kg) was added after the extrusion process along with the other micronutrients. The final concentration of curcumin was 32.9 mg/kg. The control feed was composed of the same ingredients without curcumin. After a storage of 6 months, feed composition and pH did not differ; however, the feed with curcumin showed lower protein oxidation, lipid peroxidation and higher total antioxidant capacity. After 2 months of feed production, 12 young Beagle dogs received either curcumin-containing food (n = 6) or the control diet (n = 6). The animals were fed twice a day using individual kennels. Blood samples were taken on d 1, 35 and 42. During the first 30 d of the study, the animals had natural infectious diseases that were controlled with anti-protozoals and antibiotics. Greater numbers of red blood cells were observed in dogs fed with curcumin (d 35 and 45), and there were greater numbers of white blood cells as a consequence of increased neutrophils on d 42. At the end of the experiment, a significant reduction in the number of lymphocytes was observed in dogs that ingested curcumin (d 42), suggesting an anti-inflammatory effect, manifested as a decrease in globulin levels. In the final 15 d of the experiment, the animals were clinical healthy. Higher serum levels of glucose, urea, triglycerides and cholesterol were observed in dogs fed with curcumin. Curcumin increased the activity of several antioxidant enzymes in addition to non-protein thiols and the total antioxidant capacity in the serum, consequently reducing levels of oxygen reactive species. Curcumin supplementation of dogs did not favour growth or weight gain. Neverthless, it was concluded that curcumin improved animal health, with emphasis on the stimulation of the antioxidant system and evidence of an anti-inflammatory effect.     

Differential time course of liver and kidney glucose-6 phosphatase activity during long-term fasting in rat correlates with differential time course of messenger RNA level

We have studied the role of Glc6Pase mRNA abundance in the time course of Glc6Pase activity in liver and kidney during long-term fasting in rat. Refered to the mRNA level in the fed state, Glc6Pase mRNA abundance was increased by 3.5 ± 0.5 and 3.7 ± 0.5 times (mean ± S.E.M., n = 5) in the 24 h and 48 h-fasted liver, respectively. Then, the liver Glc6Pase mRNA was decreased to the level of the fed liver after 72 and 96 h of fasting (1.0 ± 0.3 and 1.4 ± 0.3). In the kidney, Glc6Pase mRNA abundance was increased by 2.7 ± 1.0 and 5 ± 1.2 times at 24 and 48 h of fasting, respectively. Then, it plateaued at the level of the 48 h fasted kidney after 72 h and 96 h of fasting (4.5 ± 1.0 and 4.3 ± 1.0). After 24 and 48 h-refeeding, the abundance of Glc6Pase mRNA in 48 h-fasted rats was decreased to the level found in the liver and kidney of fed rats. The time course of the activity of Glc6Pase catalytic subunit during fasting and refeeding was strikingly parallel to the time course of Glc6Pase mRNA level in respective tissues. These data strongly suggest that the differential expression of Glc6Pase activity in liver and kidney in the course of fasting may be accounted for by the respective time course of mRNA abundance in both organs.Abbreviations Glc6Pase Glucose-6 phosphatase - GNG Gluconeogenesis     

Apparent influences of host-plant distribution on the structure and the genetic variability of local populations of the Purple Clay (Diarsia brunnea)

Diarsia brunnea (Lepidoptera, Heterocera, Noctuidae, Denis and Schiffermuller, 1775) is an abundant oligophagous moth occurring in the French Pyrenees. No or little influence of the forest type was found on population densities. In order to study the genetic structure of two separate moth populations in a natural forest and in a plantation, genomic fingerprinting with ISSR markers (Inter Simple Sequence Repeats) was used. The goal was to search for potential spatial structuring which could be influenced by differences in forest type.     

Essential oil of Lippia alba as a sedative and anesthetic for the sea urchin Echinometra lucunter (Linnaeus, 1758)

The sea urchin, Echinometra lucunter, is consumed in restaurants in Europe and Asia for high prices. This study evaluated the use of the essential oil of Lippia alba (EOL) as a sedative and anesthetic in this sea urchin. Different EOL concentrations were tested (50, 100 and 150 μL L^?1), in addition to a group that received ethanol and a control group. After the anesthesia and recovery from the different treatments, the coelomic fluid, gonads and intestine were collected for the analysis of oxidative stress parameters. The highest concentration tested (150 μL L^?1) was determined to be the optimal concentration for anesthesia. Results suggest that EOL improved the response to oxidative stress, since a lower level of thiobarbituric acid-reactive substances and greater superoxide dismutase and catalase activities were observed in the coelomic fluid and E. lucunter gonads, making the EOL a promising anesthetic for sea urchins.     

In the QTL region surrounding porcine MHC, gene order is conserved with human genome

On the porcine genome, the region surrounding the Major Histocompatibility Complex, also called Swine Leukocyte Antigens (SLA), is of particular interest not only owing to itq role in the control of immune response, but also because of its influence on many traits such as growth, fatness, and meat quality. To help in the identification of responsible genes, detailed comparative maps of the MHC region in mammalian species and powerful mapping tools allowing accurate ordering of genes and markers in this region are needed. In this report, we describe the use of the recently developed IMpRH radiation hybrid panel, to construct a higher density radiation hybrid map of swine Sscr 7p-q12, containing 23 additional loci. Our results show that the gene order is conserved between the two MHC-containing regions, even if an inversion is observed above the QTL region in the region containing DEK, SCA1, and EDN1 genes. The framework map produced shows that the IMpRH panel permits the ordering of genes and markers in the three MHC classes and would thus allow accurate localization of ESTs and candidate genes.