Distinct N and C cross-feeding networks in a synthetic mouse gut consortium

The complex interactions between gut microbiome and host or pathogen colonization resistance cannot solely be understood from community composition. Missing are causal relationships such as metabolic interactions among species to better understand what shapes the microbiome. Here, we focused on metabolic niches generated and occupied by the Oligo-Mouse-Microbiota consortium, a synthetic community composed of 12 members that is increasingly used as a model for the mouse gut microbiome. Combining mono-cultures and spent medium experiments with untargeted metabolomics uncovered broad metabolic diversity in the consortium, constituting a dense cross-feeding network with more than 100 pairwise interactions. Quantitative analysis of the cross-feeding network revealed distinct C and N food webs that highlight the two Bacteroidetes consortium members B. caecimuris and M. intestinale as primary suppliers of carbon, and a more diverse group as nitrogen providers. Cross-fed metabolites were mainly carboxylic acids, amino acids, and the so far not reported nucleobases. In particular the dicarboxylic acids malate and fumarate provided a strong physiological benefit to consumers, presumably as anaerobic electron acceptors. Isotopic tracer experiments validated the fate of a subset of cross-fed metabolites, in particular the conversion of the most abundant cross-fed compound succinate to butyrate. Thus, we show that this consortium is tailored to produce the anti-inflammatory metabolite butyrate. Overall, we provide evidence for metabolic niches generated and occupied by OMM members that lays a metabolic foundation to facilitate understanding of the more complex in vivo behavior of this consortium in the mouse gut.

RADIOCARBON AND ATMOSPHERIC 14CO2 PIONEER ATHOL RAFTER

ABSTRACT Direct atmospheric 14CO2 measurements began in New Zealand in 1954, initially to improve 14C as a dating tool, but quickly evolving into a method for understanding the carbon cycle. These early 14CO2 measurements immediately demonstrated the existence of an “Atom Bomb Effect,” as well as an “Industrial Effect.” These two gigantic tracer experiments have been utilized via 14CO2 measurements over the years to produce a wealth of knowledge in multiple research fields including atmospheric carbon cycle research, oceanography, soil science, and aging of post-bomb materials.

Nitrogen cycling in muddy sediments of Great Peconic Bay, USA: Seasonal N reaction balances and multi-year flux patterns

To better understand the capacity of sediments to serve as both source and sink of nitrogen (N) and to identify any evidence of evolving changes in sedimentary N cycling, N2 production, N remineralization, and N2 fixation were studied over a multi-year period (2010–2015) in bioturbated mud of Great Peconic Bay, a temperate northeastern U. S. estuary. Benthic fluxes and rates of organic matter remineralization were measured using in situ and ex situ incubations. Net annual NH+ 4, NO–3/NO–2, and N2–N fluxes (μ = 1.1, 0.03, and 1.2 mmol m –2d –1) were close to averages for comparable sedi- mentary environments from surveys of published field studies. Net N2 fluxes (by membrane inlet mass spectrometry) were influenced in different periods by temperature, oxygenation of sediment, pulsed Corg, and the activity of benthic macrofauna and benthic microalgae, although no single physical or biogeochemical variable showed a strong, direct relationship with net N2 fluxes over all sampling periods. In situ measurements sometimes showed more dynamic and higher amplitude diurnal N flux cycles than did ex situ incubations, suggesting ex situ incubations did not fully capture impacts of bioirrigation or benthic photosynthesis.15 N tracer experiments indicated anammox was < 7% of total N2 production. Acetylene reduction assays demonstrated C2 H4 production to depths ≥ 15 cm and suggested N2 fixation may have approached 25% of gross N2 production(3:1 C2 H4 : N2). Mass balances incorporating independently measured N remineralization estimates were consistent with measured levels of N2 fixation. Overall, complex balances of competing processes governed sedimentary N cycling seasonally, and N2 production dominated N2 fixation. Measured N2 fixation was consistent with constraints from N remineralization rates and net N fluxes except in episodic conditions (e. g., algal blooms). There was no indication of progressive changes in N cycling magnitudes or relative N reaction balances over the study period.

Glutamine anaplerosis is required for amino acid biosynthesis in human meningiomas

Background We postulate that meningiomas undergo distinct metabolic reprogramming in tumorigenesis and unravelling their metabolic phenotypes provide new therapeutic insights. Glutamine catabolism is key to the growth and proliferation of tumors. Here, we investigated the metabolomics of freshly resected meningiomas and glutamine metabolism in patient-derived meningioma cells. Methods 1H NMR spectroscopy of tumor tissues from 33 meningioma patients was used to differentiate the metabolite profiles of grade-I and grade-II meningiomas. Glutamine metabolism was examined using 13C/ 15N glutamine tracer, in five patient-derived meningioma cells. Results Alanine, lactate, glutamate, glutamine, and glycine were predominantly elevated only in grade-II meningiomas by 74%, 76%, 35%, 75% and 33% respectively, with alanine, and glutamine being statistically significant (p ≤ 0.02). 13C/ 15N glutamine tracer experiments revealed that both grade-I and -II meningiomas actively metabolize glutamine to generate various key carbon intermediates including alanine and proline that are necessary for the tumor growth. Also, it is shown that glutaminase (GLS1) inhibitor, CB-839 is highly effective in downregulating glutamine metabolism and decreasing proliferation in meningioma cells. Conclusion Alanine and glutamine/glutamate are mainly elevated in grade-II meningiomas. Grade-I meningiomas possess relatively higher glutamine metabolism providing carbon/nitrogen for the biosynthesis of key nonessential amino acids. GLS1 inhibitor (CB-839) would be very effective in downregulating glutamine metabolic pathways in grade-I meningiomas leading to decreased cellular proliferation.

Cyanate is a low abundance but actively cycled nitrogen compound in soil

Cyanate can serve as a nitrogen and/or carbon source for different microorganisms and as an energy source for autotrophic ammonia oxidizers. However, the extent of cyanate availability and utilisation in terrestrial ecosystems and its role in biogeochemical cycles is poorly known. Here we analyse cyanate concentrations in soils across a range of soil types, land management practices and climates. Soil cyanate concentrations were three orders of magnitude lower than ammonium or nitrate. We determined cyanate consumption in a grassland and rice paddy soil using stable isotope tracer experiments. We find that cyanate turnover was rapid and dominated by biotic processes. We estimated that in-situ cyanate production rates were similar to those associated with urea fertilizer decomposition, a major source of cyanate in the environment. We provide evidence that cyanate is actively turned over in soils and represents a small but continuous nitrogen/energy source for soil microbes.

Function and Biomarkers of the Blood-Brain Barrier in a Neonatal Germinal Matrix Haemorrhage Model

Germinal matrix haemorrhage (GMH), caused by rupturing blood vessels in the germinal matrix, is a prevalent driver of preterm brain injuries and death. Our group recently developed a model simulating GMH using intrastriatal injections of collagenase in 5-day-old rats, which corresponds to the brain development of human preterm infants. This study aimed to define changes to the blood-brain barrier (BBB) and to evaluate BBB proteins as biomarkers in this GMH model. Regional BBB functions were investigated using blood to brain 14C-sucrose uptake as well as using biotinylated BBB tracers. Blood plasma and cerebrospinal fluids were collected at various times after GMH and analysed with ELISA for OCLN and CLDN5. The immunoreactivity of BBB proteins was assessed in brain sections. Tracer experiments showed that GMH produced a defined region surrounding the hematoma where many vessels lost their integrity. This region expanded for at least 6 h following GMH, thereafter resolution of both hematoma and re-establishment of BBB function occurred. The sucrose experiment indicated that regions somewhat more distant to the hematoma also exhibited BBB dysfunction; however, BBB function was normalised within 5 days of GMH. This shows that GMH leads to a temporal dysfunction in the BBB that may be important in pathological processes as well as in connection to therapeutic interventions. We detected an increase of tight-junction proteins in both CSF and plasma after GMH making them potential biomarkers for GMH.