Armenia and Belarus: caught between the EU's and Russia's conditionalities?

This article looks into Armenia's and Belarus’ engagement with the European Union's (EU) and Russia's conditionalities, the two EU Eastern Partnership (EaP) countries that are also members of the Russia-led Eurasian Economic Union (EAEU). While paying attention to political, economic (including energy and technical) as well as security dimensions of the EU's and Russia's approaches, as proposed in the present special section, the article demonstrates that the conditionalities extended by the EU and Russia to the two countries in question have differed. In their turn, Armenia and Belarus have reacted differently to Russia's and the EU's conditionalities. Against the backdrop of the changing significance ascribed to both the EU's and Russia's policies towards their common neighbourhood since the 1990s, the present contribution identifies and analyses factors that account for the diverging positions of Armenia and Belarus, including the type of regime, the geopolitical considerations, the stakes in the economic and energy spheres and the predisposition to integration. The article shows that in the resulting complex context, Armenia and Belarus have been able to influence the shape and content of the EU's and Russia's conditionalities, although in a different way and to a different extent.Ministry of Education and Science (UID/CPO/ 00758/2013

Foaming and air-water interfacial characteristics of solutions containing both gluten hydrolysate and egg white protein

Enzymatically hydrolyzed wheat gluten can be a viable alternative for traditional animal-based foam stabilizing proteins in food systems. Gluten hydrolysates (GHs) can be considered for (partially) replacing surface-active food proteins such as those of egg white (EW). We here studied the foaming and air-water (A-W) interfacial characteristics of mixed GH + EW protein solutions. GH solutions had much higher (P < 0.05) foaming capacities than EW solutions, while the latter had much higher (P < 0.05) foam stability than the former. When only one sixth of EW proteins was replaced by GHs, the foaming capacity of the mixtures was as high as or higher than that of the GH solutions. Furthermore, when half of the EW protein was replaced by GH, the mixtures still had high foam stability. It thus seems that both GH and EW proteins contribute positively to the foaming characteristics of the mixtures. However, measurements of the early stages of diffusion to and adsorption at the interface, plus measurements of surface dilatational moduli at the interface, both suggested that the adsorbed protein film consists primarily of GHs rather than of EW proteins. Nonetheless, FS was higher when EW proteins were present. Mixed GH + EW solutions have a higher resistance to coalescence than GH solutions. Therefore, it is hypothesized that EW proteins form a secondary protein layer below the A-W interface which is maintained by interactions with adsorbed GH constituents, thereby providing bubbles with an additional resistance to coalescence

The European Union, Russia and the Eastern region: The analytics of government for sustainable cohabitation

This article applies the Foucauldian premise of governmentality and the analytics of government framework to demonstrate how exclusive modalities of power – of the European Union (EU) and Russia – and their competing rationalities relate, intersect and become, counter-intuitively, inextricable in their exercise of governance over the eastern neighbourhood. This particular approach focuses on power as a process to gauge the prospects for compatibility and cohabitation between the EU and Russia. Using original primary evidence, this article contends that cohabitation between these two exclusive power modalities is possible and even inevitable, if they were to legitimise their influence over the contested eastern region. It also exposes a fundamental flaw in the existing power systems, as demonstrated so vividly in the case of Ukraine – that is, a neglect for the essential value of freedom in fostering subjection to one’s authority, and the role of ‘the other’ in shaping the EU–Russian power relations in the contested regio

Tailored ß-Cyclodextrin Blocks the Translocation Pores of Binary Exotoxins from C. Botulinum and C. Perfringens and Protects Cells from Intoxication

International audienceBackgroundClostridium botulinum C2 toxin and Clostridium perfringens iota toxin are binary exotoxins, which ADP-ribosylate actin in the cytosol of mammalian cells and thereby destroy the cytoskeleton. C2 and iota toxin consists of two individual proteins, an enzymatic active (A-) component and a separate receptor binding and translocation (B-) component. The latter forms a complex with the A-component on the surface of target cells and after receptor-mediated endocytosis, it mediates the translocation of the A-component from acidified endosomal vesicles into the cytosol. To this end, the B-components form heptameric pores in endosomal membranes, which serve as translocation channels for the A-components.Here we demonstrate that a 7-fold symmetrical positively charged ß-cyclodextrin derivative, per-6-S-(3-aminomethyl)benzylthio-ß-cyclodextrin, protects cultured cells from intoxication with C2 and iota toxins in a concentration-dependent manner starting at low micromolar concentrations. We discovered that the compound inhibited the pH-dependent membrane translocation of the A-components of both toxins in intact cells. Consistently, the compound strongly blocked transmembrane channels formed by the B-components of C2 and iota toxin in planar lipid bilayers in vitro. With C2 toxin, we consecutively ruled out all other possible inhibitory mechanisms showing that the compound did not interfere with the binding of the toxin to the cells or with the enzyme activity of the A-component.Conclusions/SignificanceThe described ß-cyclodextrin derivative was previously identified as one of the most potent inhibitors of the binary lethal toxin of Bacillus anthracis both in vitro and in vivo, implying that it might represent a broad-spectrum inhibitor of binary pore-forming exotoxins from pathogenic bacteria

Acetate Kinase Isozymes Confer Robustness in Acetate Metabolism

Acetate kinase (ACK) (EC no: 2.7.2.1) interconverts acetyl-phosphate and acetate to either catabolize or synthesize acetyl-CoA dependent on the metabolic requirement. Among all ACK entries available in UniProt, we found that around 45% are multiple ACKs in some organisms including more than 300 species but surprisingly, little work has been done to clarify whether this has any significance. In an attempt to gain further insight we have studied the two ACKs (AckA1, AckA2) encoded by two neighboring genes conserved in Lactococcus lactis (L. lactis) by analyzing protein sequences, characterizing transcription structure, determining enzyme characteristics and effect on growth physiology. The results show that the two ACKs are most likely individually transcribed. AckA1 has a much higher turnover number and AckA2 has a much higher affinity for acetate in vitro. Consistently, growth experiments of mutant strains reveal that AckA1 has a higher capacity for acetate production which allows faster growth in an environment with high acetate concentration. Meanwhile, AckA2 is important for fast acetate-dependent growth at low concentration of acetate. The results demonstrate that the two ACKs have complementary physiological roles in L. lactis to maintain a robust acetate metabolism for fast growth at different extracellular acetate concentrations. The existence of ACK isozymes may reflect a common evolutionary strategy in bacteria in an environment with varying concentrations of acetate

The antimicrobial effects of the alginate oligomer OligoG CF-5/20 are independent of direct bacterial cell membrane disruption

Concerns about acquisition of antibiotic resistance have led to increasing demand for new antimicrobial therapies. OligoG CF-5/20 is an alginate oligosaccharide previously shown to have antimicrobial and antibiotic potentiating activity. We investigated the structural modification of the bacterial cell wall by OligoG CF-5/20 and its effect on membrane permeability. Binding of OligoG CF-5/20 to the bacterial cell surface was demonstrated in Gram-negative bacteria. Permeability assays revealed that OligoG CF-5/20 had virtually no membrane-perturbing effects. Lipopolysaccharide (LPS) surface charge and aggregation were unaltered in the presence of OligoG CF-5/20. Small angle neutron scattering and circular dichroism spectroscopy showed no substantial change to the structure of LPS in the presence of OligoG CF-5/20, however, isothermal titration calorimetry demonstrated a weak calcium-mediated interaction. Metabolomic analysis confirmed no change in cellular metabolic response to a range of osmolytes when treated with OligoG CF-5/20. This data shows that, although weak interactions occur between LPS and OligoG CF-5/20 in the presence of calcium, the antimicrobial effects of OligoG CF-5/20 are not related to the induction of structural alterations in the LPS or cell permeability. These results suggest a novel mechanism of action that may avoid the common route in acquisition of resistance via LPS structural modification

Enterobactin-Mediated Delivery of β-Lactam Antibiotics Enhances Antibacterial Activity against Pathogenic Escherichia coli

The design, synthesis, and characterization of enterobactin–antibiotic conjugates, hereafter Ent-Amp/Amx, where the β-lactam antibiotics ampicillin (Amp) and amoxicillin (Amx) are linked to a monofunctionalized enterobactin scaffold via a stable poly(ethylene glycol) linker are reported. Under conditions of iron limitation, these siderophore-modified antibiotics provide enhanced antibacterial activity against Escherichia coli strains, including uropathogenic E. coli CFT073 and UTI89, enterohemorrhagic E. coli O157:H7, and enterotoxigenic E. coli O78:H11, compared to the parent β-lactams. Studies with E. coli K-12 derivatives defective in ferric enterobactin transport reveal that the enhanced antibacterial activity observed for this strain requires the outer membrane ferric enterobactin transporter FepA. A remarkable 1000-fold decrease in minimum inhibitory concentration (MIC) value is observed for uropathogenic E. coli CFT073 relative to Amp/Amx, and time-kill kinetic studies demonstrate that Ent-Amp/Amx kill this strain more rapidly at 10-fold lower concentrations than the parent antibiotics. Moreover, Ent-Amp and Ent-Amx selectively kill E. coli CFT073 co-cultured with other bacterial species such as Staphylococcus aureus, and Ent-Amp exhibits low cytotoxicity against human T84 intestinal cells in both the apo and iron-bound forms. These studies demonstrate that the native enterobactin platform provides a means to effectively deliver antibacterial cargo across the outer membrane permeability barrier of Gram-negative pathogens utilizing enterobactin for iron acquisition.Pacific Southwest Regional Center of Excellence for Biodefense and Emerging Infectious DiseaseKinship Foundation. Searle Scholars ProgramMassachusetts Institute of Technology. Department of Chemistr

Expert consensus document: The International Scientific Association for Probiotics and Prebiotics (ISAPP) consensus statement on the definition and scope of prebiotics

In December 2016, a panel of experts in microbiology, nutrition and clinical research was convened by the International Scientific Association for Probiotics and Prebiotics to review the definition and scope of prebiotics. Consistent with the original embodiment of prebiotics, but aware of the latest scientific and clinical developments, the panel updated the definition of a prebiotic: a substrate that is selectively utilized by host microorganisms conferring a health benefit. This definition expands the concept of prebiotics to possibly include non-carbohydrate substances, applications to body sites other than the gastrointestinal tract, and diverse categories other than food. The requirement for selective microbiota-mediated mechanisms was retained. Beneficial health effects must be documented for a substance to be considered a prebiotic. The consensus definition applies also to prebiotics for use by animals, in which microbiota-focused strategies to maintain health and prevent disease is as relevant as for humans. Ultimately, the goal of this Consensus Statement is to engender appropriate use of the term ‘prebiotic’ by relevant stakeholders so that consistency and clarity can be achieved in research reports, product marketing and regulatory oversight of the category. To this end, we have reviewed several aspects of prebiotic science including its development, health benefits and legislation