A number of approaches could be used

A number of approaches could be used to further improve the efficiency of our IFN-γ release assay: addition to the stimulated blood culture of recombinant bovine IL-12 and IL-2 (Jungersen et al., 2012) and IL-7 alone or in combination with IL-12 (Plain et al., 2012) and neutralization of IL-10 to enhance production of IFN-γ by Th1 cells (Buza et al., 2004; Mikkelsen et al., 2012).
Finally, it would be interesting to test these Map specific antigens in antigen cocktails, as it was demonstrated for the diagnosis of bovine tuberculosis, using IFN-γ whole-blood assays (Aagaard et al., 2010) and serological multiplex immunoassays (Whelan et al., 2008).

K. Dernivoix was supported by the Belgian Federal Public Service of Health (Convention RT-07/6 PARATUB 2008-2012). V. Roupie held a FRIA scholarship and was further supported by the Belgian Federal Public Service of Health (Convention RT-07/6 PARATUB, 2008-2012). S. Viart held a FRIA scholarship. This work was partially funded through a grant from the Walloon Region (J.-J.L. and R.W.). We are very grateful to ARSIA, DGZ, veterinarians and farmers for their assistance, and to D. Flament and M. Den Haerynck for their excellent technical assistance.

Salmonella has emerged as one of the most common pathogens and the leading cause of foodborne illnesses worldwidely (Zhang et al., 2017). S. enteritidis can cause symptoms of food poisoning, such as fever, diarrhea, and vomiting, following intake of contaminated poultry products, and can cause death of poultry, resulting in serious economic losses (Lin et al., 2002; Zhang et al., 2016). In some cases, Avasimibe have been overused to prevent and cure S. enteritidis infection, leading to increasing drug resistance to S. enteritidis and serious impact on food safety (Shen et al., 2015). Given the limitations of available methods for control of S. enteritidis, research into the genetic mechanism of innate immunity and its potential to control the spread and prevalence of S. enteritidis has attracted increasing attention in recent years, especially the difference in resistance to S. enteritidis in different breeds of chickens.
Toll-like receptors are a family of innate immunity receptorsPlease add the reference \”(Akira et al., 2001)\”.. These receptors can identify pathogen-associated molecular patterns, and trigger signal transduction resulting in the release of inflammatory mediators including cytokines, producing a series of immune and pathological reactions that result in immune protection (Akira and Takeda, 2004). Toll-like receptor 4 (TLR4) is one of the most important Toll-like receptors for identification of Salmonella in mammals (Weiss et al., 2004). It is also the main transmembrane receptor for transduction of endotoxin signals, and lipopolysaccharide (LPS) is the major pathogen-associated molecular pattern (PAMP) of Salmonella (Mansell et al., 2001; Norata et al., 2005). TLR4 mediates the MyD88 and TRIF signalling pathways. The MyD88 signalling pathway includes the genes MyD88, NF-κB, as well as lipopolysaccharide-induced TNF-α factor (LITAF) and IL-8 (CXCLi2) (Keestra and van Putten, 2008; Larson et al., 2008; Chaussé et al., 2011). The MyD88 signalling pathway plays a key role in the regulation of innate and adaptive immunity (Zhou et al., 2005; Loures et al., 2011). The TRIF signalling pathway mainly includes TRIF, IRF3 and INFβ. This pathway is also known as the MyD88-independent signalling pathway, and it mainly mediates involvement of type I IFN in the immune response (Yamamoto et al., 2003; Ramakrishnan et al., 2015).
DNA methylation is an important component of epigenetic regulation, and it plays a key role in gene expression and regulation (Brown et al., 2012). Members of the Toll-like receptor family, especially TLR4 and TLR2, have been intensively investigated for their role in promoter region methylation and inflammatory reactions (De Oliveira et al., 2011; Diesel et al., 2012; Benakanakere et al., 2015). Few reports have been published on the relationship between inflammation and TLR4 methylation caused by stimulation of Salmonella in mice (Gou et al., 2012). However, the effects of TLR4 methylation on the expression of genes in the MyD88 and TRIF signalling pathways in poultry, especially chickens following Salmonella infection have not been reported. In addition, no reports have been published on DNA methylation of TRIF, a key gene in the TRIF signalling pathway. Gou et al. (2012), studying S. enteritidis-resistant and −susceptible groups of the SPF chickens, report that expression of TLR4 in susceptible chickens may be caused by excessive methylation (Gou et al., 2012).Li et al. (2010) find that the MyD88 pathway plays an important role in infection of Salmonella Pullorum in chickens (Li et al., 2010). Karnati et al. (2015) show that LPS can stimulate activation of the MyD88 and TRIF signalling pathways in peripheral blood of chickens. Sadeyen et al. (2006) report that the innate immunity molecules, including TLR4, are constitutively extremely expressed in resistant adult birds and thus suggesting they play an important role in the host to fight against Salmonella colonization. Redmond et al. (2009) compare mRNA expression of TLR4 and genes encoding inflammatory-related cytokines after S. enteritidis infection in three breeds of chickens and found significant differences between Fayoumi chicken strains and white feather chickens and among others. Recently, Herrmann et al. (2015) show that TLR4 and MyD88 gene expressions differ significantly at different times post-stimulation in response to LPS stimulation and three chicken lines have different expression levels of TLR4 in response to LPS. These reveal that there are genetic differences in this immune pathway. Under stimulation with S. enteritidis, differences in genetic background may lead to differences in methylation levels of TLR4 and the expression of its downstream genes. In the present study, we investigate the effects of S. enteritidis infection on Tibetan chickens and DaHeng chickens, and methylation of TLR4 and TRIF, as well as the expression differences of genes and proteins in the MyD88 and TRIF signalling pathway, to determine the genetic mechanism underlying S. enteritidis resistance in both two breeds of chickens.

Although we did not find a relationship between expression of

Although we did not find a relationship between expression of Bcl-2 proteins and resistance of PDAC cell lines to VSV-ΔM51-mediated oncolysis, further studies are also needed to determine the influence of these or other regulators downstream of caspase 8 and 9 such as inhibitors of apoptosis (IAPs) (Hamacher et al., 2008). Our future studies will also examine the role of other types of cell death in VSV infected PDACs, particularly autophagy as VSV has already been reported to induce autophagy in different types of cancer Avasimibe (Schache et al., 2009).

Materials and methods

We thank Eric Hastie and Marcela Cataldi for critical comments on the manuscript. This work was supported by Grant1R15CA167517-01 (to V.Z.G.) from the National Institute of Health, USA (National Cancer Institute).

Rabies is an important zoonotic disease that is estimated to kill at least 60 000 people worldwide each year (Martinez, 2000). It is caused by the rabies virus, a non-segmented, negative-strand lyssavirus in the Rhabdoviridae family. The majority of rabies deaths occurs from rabid dog bites in developing countries where medical resources are limited and/or a lack of awareness about the risk of RABV exposure exists (Knobel et al., 2005).
PEP readily prevents development of clinical rabies and death when given soon after exposure to a rabid animal (Franka et al., 2009). Unfortunately, because many do not have access to proper medical care, whether for financial or logistical reasons, or may not recognize a potential RABV exposure, prompt administration of rabies PEP to at-risk individuals is not always achieved (Joseph et al., 2013). In these cases RABV enters the brain, clinical rabies develops and the infected individuals almost invariably die. Therefore, there is a great need to decrease RABV infection in dogs (and other reservoirs) and to develop new therapeutics to successfully extend the window of successful post-exposure treatment from days to perhaps weeks or months. Live-attenuated vaccines are promising candidates to fulfill both these needs. They can be produced economically and induce long-lasting immunity with a single dose. Furthermore, superinfection with attenuated RABV has successfully protected animals exposed to lethal RABV.
Early superinfection experiments in dogs showed protection from intramuscular (i.m.) injection of a lethal RABV by intrathecal injection of an attenuated RABV (Baer et al., 1975). Importantly, inactivated RABV could not protect dogs from death in these experiments. Since that time, other animal model experiments have demonstrated the ability of live-attenuated RABV to protect against infection with lethal wild-type RABV (Franka et al., 2009; Faber et al., 2009; Li et al., 2012; Schutsky et al., 2013). These data indicate that treatment with attenuated RABV has the potential to protect individuals who would otherwise succumb to rabies.
While questions of reverse mutations and safety persist, the effectiveness of GAS RABV variants, with two attenuating mutations in their glycoproteins, has been well established (Faber et al., 2009; Li et al., 2012; Schutsky et al., 2013). Much of the success of the GAS variants has been attributed to rapid induction of virus neutralizing antibody (VNA) (Faber et al., 2009; Schutsky et al., 2013). In order to be an effective late PET vector, the virus should initiate an immune response much faster than the typical time that is required to induce a robust VNA response. We recently showed that the addition of IFNγ to a pathogenic RABV backbone highly attenuates the virus and enhances the innate immune response, specifically through induction of type I interferons (Barkhouse et al., 2014). Therefore, we constructed two new IFNγ-expressing GAS vectors, GASγ and GASγGAS, to determine if IFNγ expression by a highly attenuated vaccine vector could increase its safety profile and/or its efficacy as pre- and post-exposure treatment.

Lipid sample preparation Krill samples used for lipid

2.4. Lipid sample preparation
Krill samples used for lipid class analysis were freeze-dried to calculate dry mass, then extracted overnight using a modified Bligh and Dyer (1959) one-phase methanolachloroformawater extraction (2:1:0.8 v/v/v). The phases were separated by addition of chloroformawater (final solvent ratio, 1:1:0.9 v/v/v methanolachloroformawater). The total solvent extract (TSE) was concentrated using rotary evaporation at 40uC then stored in chloroform in 1.5uml glass vials with Teflon-lined caps at 20uC until analysis. Total lipid content was determined gravimetrically after evaporation of CHCl3 under a stream of nitrogen until dry.
For lipid class composition, an aliquot of the total lipid extract was analysed using an Iatroscan MK V TH10 thin-layer chromatography-flame ionisation detector (TLC-FID) analyser (Tokyo, Japan) (Ackman, 1981uanduVolkman and Nichols, 1991). The FID was calibrated for each lipid class (phosphatidylcholine, cholesterol, cholesteryl oleate, oleic acid, squalene, TAG (derived from fish oil), wax ester (derived from orange roughy, Hoplostethus atlanticus, oil) and diacylglyceryl ether (DAGE; derived from shark liver oil; 0.1a10uμg range)). Peaks were quantified using SIC-480 Scientific Software. TLC-FID results are generally reproducible to +/10% of individual class abundances (Volkman and Nichols, 1991).
An aliquot of the total lipid extract was trans-methylated to produce fatty Avasimibe methyl esters as described in Virtue et al. (1996). Samples were analysed by gas chromatography (GC) using an Agilent Technologies 7890B GC (Palo Alto, California, USA) filtered with an Equity™-1 fused silica capillary column (15um0.1umm i.d., 0.1uμm film thickness), an FID, a split/splitless injector and an Agilent Technologies 7683B Series auto sampler and injector. Helium was the carrier gas. Samples were injected in splitless mode at an oven temperature of 120uC. After injection, the oven temperature was raised to 270uC at 10uCumina1 and finally to 310uC at 5uCumina1. GC results are subject to an error of 5% of individual component area. GCamass spectrometric (GCaMS) analyses were performed on a Finnigan Thermoquest DSQ GCamass spectrometer fitted with an on-column injector using Thermoquest Xcalibur software (Austin, Texas, USA).
2.5. Instantaneous growth rate (IGR)
Krill were used for growth experiments using the IGR method (Virtue et al., 2010). Immediately after capture, live krill were maintained individually at 0.5uC. Jars were checked for moults at 24uh intervals. Growth increments (% growth per moult) were calculated by measuring the length of both left and right uropod exopodites on the fresh krill and their discarded exoskeletons using a digital microscope camera equipped with image analysis software. Growth increments (GI) were calculated using the formula:GI=100(LULMLM)+(RURMRM)2where LU is the total length of the animal;s left uropod, LM is the length of the moulted left uropod, RU is the length of the animal;s right uropod, and RM is the length of the moulted right uropod. Where one uropod was damaged, GI was calculated based on the undamaged side only. If both left and right uropods were damaged, the animal was excluded from the study.

Estimation of specific carbon emissions resulting from electrical consumption is

Estimation of specific carbon emissions resulting from electrical consumption is determined considering that 1 kWh = 800 g CO2[26]. Energy consumption of each process was measured using an electrical meter (Cost Control, La Crosse Technology, France).
3. Results and discussion
3.1. Comparison of extraction processes in terms of extraction yield
Extraction yields obtained by heat reflux extraction (HRE), maceration, ultrasound and microwave assisted extraction are presented in Fig. 3. Regarding HRE extractions, it Avasimibe can be identified that most of the extraction is achieved within 30 min. Increasing the duration of HRE up to 5 h does not lead to a drastic increase of the extraction yield (20% against 19% for 5 h and 30 min extraction duration respectively). Moreover, adding a preliminary step of homogenization prior to HRE does not improve significantly the yield (18.8 ± 0.2% and 19.0 ± 0.5% respectively). Maceration at 40 °C for 30 min results in a much lower extraction yield (10.0 ± 0.3%). These differences in yield between HRE and maceration are renin attributed to the temperature difference during extraction.