A Continuous Battle for Host-Derived Glycans Between a Mucus Specialist and a Glycan Generalist in vitro and in vivo

The human gastrointestinal tract is colonized by a diverse microbial community, which plays a crucial role in human health. In the gut, a protective mucus layer that consists of glycan structures separates the bacteria from the host epithelial cells. These host-derived glycans are utilized by bacteria that have adapted to this specific compound in the gastrointestinal tract. Our study investigated the close interaction between two distinct gut microbiota members known to use mucus glycans, the generalist Bacteroides thetaiotaomicron and the specialist Akkermansia muciniphila in vitro and in vivo. The in vitro study, in which mucin was the only nutrient source, indicated that B. thetaiotaomicron significantly upregulated genes coding for Glycoside Hydrolases (GHs) and mucin degradation activity when cultured in the presence of A. muciniphila. Furthermore, B. thetaiotaomicron significantly upregulated the expression of a gene encoding for membrane attack complex/perforin (MACPF) domain in co-culture. The transcriptome analysis also indicated that A. muciniphila was less affected by the environmental changes and was able to sustain its abundance in the presence of B. thetaiotaomicron while increasing the expression of LPS core biosynthesis activity encoding genes (O-antigen ligase, Lipid A and Glycosyl transferases) as well as ABC transporters. Using germ-free mice colonized with B. thetaiotaomicron and/or A. muciniphila, we observed a more general glycan degrading profile in B. thetaiotaomicron while the expression profile of A. muciniphila was not significantly affected when colonizing together, indicating that two different nutritional niches were established in mice gut. Thus, our results indicate that a mucin degrading generalist adapts to its changing environment, depending on available carbohydrates while a mucin degrading specialist adapts by coping with competing microorganism through upregulation of defense related genes.Peer reviewe

Microbial Metabolic Networks at the Mucus Layer Lead to Diet-Independent Butyrate and Vitamin B12 Production by Intestinal Symbionts

Peer reviewe

Deciphering the trophic interaction between Akkermansia muciniphila and the butyrogenic gut commensal Anaerostipes caccae using a metatranscriptomic approach

Host glycans are paramount in regulating the symbiotic relationship between humans and their gut bacteria. The constant flux of host-secreted mucin at the mucosal layer creates a steady niche for bacterial colonization. Mucin degradation by keystone species subsequently shapes the microbial community. This study investigated the transcriptional response during mucin-driven trophic interaction between the specialised mucin-degrader Akkermansia muciniphila and a butyrogenic gut commensal Anaerostipes caccae. A. muciniphila monocultures and co-cultures with non-mucolytic A. caccae from the Lachnospiraceae family were grown anaerobically in minimal media supplemented with mucin. We analysed for growth, metabolites (HPLC analysis), microbial composition (quantitative reverse transcription PCR), and transcriptional response (RNA-seq). Mucin degradation by A. muciniphila supported the growth of A. caccae and concomitant butyrate production predominantly via the acetyl-CoA pathway. Differential expression analysis (DESeq 2) showed the presence of A. caccae induced changes in the A. muciniphila transcriptional response with increased expression of mucin degradation genes and reduced expression of ribosomal genes. Two putative operons that encode for uncharacterised proteins and an efflux system, and several two-component systems were also differentially regulated. This indicated A. muciniphila changed its transcriptional regulation in response to A. caccae. This study provides insight to understand the mucin-driven microbial ecology using metatranscriptomics. Our findings show that the expression of mucolytic enzymes by A. muciniphila increases upon the presence of a community member. This could indicate its role as a keystone species that supports the microbial community in the mucosal environment by increasing the availability of mucin sugars.Peer reviewe

Akkermansia muciniphila uses human milk oligosaccharides to thrive in the early life conditions in vitro

Akkermansia muciniphila is a well-studied anaerobic bacterium specialized in mucus degradation and associated with human health. Because of the structural resemblance of mucus glycans and free human milk oligosaccharides (HMOs), we studied the ability of A. muciniphila to utilize human milk oligosaccharides. We found that A. muciniphila was able to grow on human milk and degrade HMOs. Analyses of the proteome of A. muciniphila indicated that key-glycan degrading enzymes were expressed when the bacterium was grown on human milk. Our results display the functionality of the key-glycan degrading enzymes (alpha -l-fucosidases, beta -galactosidases, exo-alpha -sialidases and beta -acetylhexosaminidases) to degrade the HMO-structures 2 ' -FL, LNT, lactose, and LNT2. The hydrolysation of the host-derived glycan structures allows A. muciniphila to promote syntrophy with other beneficial bacteria, contributing in that way to a microbial ecological network in the gut. Thus, the capacity of A. muciniphila to utilize human milk will enable its survival in the early life intestine and colonization of the mucosal layer in early life, warranting later life mucosal and metabolic health.Peer reviewe

Operation and Utilisation of the High Flux Reactor: Annual Report 2013

The High Flux Reactor (HFR) at Petten is managed by the Institute for Energy and Transport (IET) of the European Commission's Joint Research Centre (JRC) and operated by the Nuclear Research and consultancy Group (NRG) which is also the licence holder and responsible for its commercial activities. The High Flux Reactor (HFR) operates at 45 MW and is of the tank-in-pool type, light water cooled and moderated. It is one of the most powerful multi-purpose materials testing reactors in the world and one of the world's leaders in target irradiation for the production of medical radioisotopes.JRC.F.4-Innovative Technologies for Nuclear Reactor Safet

Operation and Utilisation of the High Flux Reactor - Annual Report 2012

The High Flux Reactor (HFR) at Petten is managed by the Institute for Energy and Transport (IET) of the European Commission's Joint Research Centre (JRC) and operated by the Nuclear Research and consultancy Group (NRG) which is also the licence holder and responsible for its commercial activities. The High Flux Reactor (HFR) operates at 45 MW and is of the tank-in-pool type, light water cooled and moderated. It is one of the most powerful multi-purpose materials testing reactors in the world and one of the world's leaders in target irradiation for the production of medical radioisotopes.JRC.F.4-Nuclear Reactor Integrity Assessment and Knowledge Managemen

Operation and Utilisation of the High Flux Reactor: Annual Report 2015

The High Flux Reactor (HFR) at Petten is managed by the Institute for Energy and Transport (IET) of the European Commission's Joint Research Centre (JRC) and operated by the Nuclear Research and consultancy Group (NRG) which is also the licence holder and responsible for its commercial activities. The High Flux Reactor (HFR) operates at 45 MW and is of the tank-in-pool type, light water cooled and moderated. It is one of the most powerful multi-purpose materials testing reactors in the world and one of the world's leaders in target irradiation for the production of medical radioisotopes.JRC.G.I.4-Nuclear Reactor Safety and Emergency Preparednes

Measurement of residual stresses in surrogate coated nuclear fuel particles using ring-core focussed ion beam digital image correlation

Coated fuel particles, most commonly tri-structural isotropic (TRISO), are intended for application in several designs of advanced nuclear reactors. A complete understanding of the residual stresses and local properties of these particles through their entire lifecycle is required to inform fuel element manufacturing, reactor operation, accident scenarios, and reprocessing. However, there is very little experimental data available in the literature on the magnitude of residual stresses in the individual coating layers of these particles. This work applies ring-core focussed ion beam milling combined with digital image correlation analysis (FIB-DIC) to cross-sections of TRISO and pyrolytic carbon coatings in surrogate coated fuel particles to evaluate the residual stresses. Tensile residual hoop stresses are identified in both pyrolytic carbon layers, while silicon carbide experiences a compressive residual hoop stress. Note that these residual stresses, which were not accounted for in the models reported in open literature, have magnitudes comparable to the stresses predicted to arise in real fuel particles during service. A 2D linear-elastic continuum-based finite element analysis has been conducted to investigate the stress relaxation phenomena caused by sectioning stressed coatings on spherical particles. The FIB-DIC method established here is independent of radiation defects and can be applied to irradiated TRISO particles to retrieve first-hand information regarding the residual stress evolution during service

Clustered mutations in the <i>GRIK2</i> kainate receptor subunit gene underlie diverse neurodevelopmental disorders

Kainate receptors (KARs) are glutamate-gated cation channels with diverse roles in the central nervous system. Bi-allelic loss of function of the KAR-encoding gene GRIK2 causes a nonsyndromic neurodevelopmental disorder (NDD) with intellectual disability and developmental delay as core features. The extent to which mono-allelic variants in GRIK2 also underlie NDDs is less understood because only a single individual has been reported previously. Here, we describe an additional eleven individuals with heterozygous de novo variants in GRIK2 causative for neurodevelopmental deficits that include intellectual disability. Five children harbored recurrent de novo variants (three encoding p.Thr660Lys and two p.Thr660Arg), and four children and one adult were homozygous for a previously reported variant (c.1969G&gt;A [p.Ala657Thr]). Individuals with shared variants had some overlapping behavioral and neurological dysfunction, suggesting that the GRIK2 variants are likely pathogenic. Analogous mutations introduced into recombinant GluK2 KAR subunits at sites within the M3 transmembrane domain (encoding p.Ala657Thr, p.Thr660Lys, and p.Thr660Arg) and the M3-S2 linker domain (encoding p.Ile668Thr) had complex effects on functional properties and membrane localization of homomeric and heteromeric KARs. Both p.Thr660Lys and p.Thr660Arg mutant KARs exhibited markedly slowed gating kinetics, similar to p.Ala657Thr-containing receptors. Moreover, we observed emerging genotype-phenotype correlations, including the presence of severe epilepsy in individuals with the p.Thr660Lys variant and hypomyelination in individuals with either the p.Thr660Lys or p.Thr660Arg variant. Collectively, these results demonstrate that human GRIK2 variants predicted to alter channel function are causative for early childhood development disorders and further emphasize the importance of clarifying the role of KARs in early nervous system development.</p