Antibodies Set Boundaries Limiting Microbial Metabolite Penetration and the Resultant Mammalian Host Response.

Although the mammalian microbiota is well contained within the intestine, it profoundly shapes development and metabolism of almost every host organ. We questioned the range and depth of microbial metabolite penetration into the host, and how this is modulated by intestinal immunity. Chemically identical microbial and host metabolites were distinguished by stable isotope tracing from 13C-labeled live non-replicating Escherichia coli, differentiating 12C host isotopes with high-resolution mass spectrometry. Hundreds of endogenous microbial compounds penetrated 23 host tissues and fluids after intestinal exposure: subsequent 12C host metabolome signatures included lipidemia, reduced glycolysis, and inflammation. Penetrant bacterial metabolites from the small intestine were rapidly cleared into the urine, whereas induced antibodies curtailed microbial metabolite exposure by accelerating intestinal bacterial transit into the colon where metabolite transport mechanisms are limiting. Pervasive penetration of microbial molecules can cause extensive host tissue responses: these are limited by immune and non-immune intestinal mucosal adaptations to the microbiota.

Phytostabilization of trace elements and 13C isotope composition of Atriplex atacamensis Phil. cultivated in mine tailings treated with organic amendments.

Mining generates large quantities of mineral processing wastes that are typically stored in mine tailings (MT) ponds. Long-term exposure of the surrounding areas to the material from the tailings ponds has been reported to have adverse effects on both human health and the environment. The purpose of this study was to evaluate the ability of Atriplex atacamensis Phil. to phytostabilize metals (Cu, Fe, Mn, and Zn) and sulfur (S) when grown directly on mine tailings with and without compost (C) and humic substance (HS). The stress status of A. atacamensis Phil. was also evaluated through the 13C isotopic composition of bulk leaves. A 120-day greenhouse experiment was conducted and three treatments were evaluated: (i) MT without any amendments (control), (ii) MT + C (dose: 89 ton ha-1), and (iii) MT + HS (0.72 ton ha-1). Mine tailings material exhibited low salinity, alkaline pH, high extractable S-SO4 concentrations, and low fertility; total Fe, Mn, and Zn concentrations were within the reference range for mine tailings, but total Cu concentrations were high at 1860 ± 236 mg kg-1. The HS had higher pH, EC, CEC, and available concentrations of N, P, and K than compost, while S-SO4 concentrations were similar in both amendments. 13C NMR analysis showed that the HS contained more alkyl, aromatic, and phenolic groups, while the compost was dominated by O-alkyl and carboxyl groups. At the end of the experiment, the MT + C treatment achieved a significant decrease in Cu, Fe, and Mn concentrations in the roots and aboveground parts of A. atacamensis Phil. and an increase in Zn values in both tissues. Both amendments increased the sulfur content in the aboveground parts, while metal concentrations under the HS treatment proved similar to control. Furthermore, the δ13CV-PDB values obtained in this study indicate that the organic amendments did not cause additional physiological stress to the plants compared to the MT treatment. Overall, A. atacamensis Phil. was shown to have the ability to phytostabilize metals and sulfur, making it a potential candidate species for in situ evaluation of the phytostabilization process on mine tailings.

IDSL.IPA Characterizes the Organic Chemical Space in Untargeted LC/HRMS Data Sets.

Generating comprehensive and high-fidelity metabolomics data matrices from LC/HRMS data remains to be extremely challenging for population-scale large studies (n > 200). Here, we present a new data processing pipeline, the Intrinsic Peak Analysis (IDSL.IPA) R package (https://ipa.idsl.me), to generate such data matrices specifically for organic compounds. The IDSL.IPA pipeline incorporates (1) identifying potential 12C and 13C ion pairs in individual mass spectra; (2) detecting and characterizing chromatographic peaks using a new sensitive and versatile approach to perform mass correction, peak smoothing, baseline development for local noise measurement, and peak quality determination; (3) correcting retention time and cross-referencing peaks from multiple samples by a dynamic retention index marker approach; (4) annotating peaks using a reference database of m/z and retention time; and (5) accelerating data processing using a parallel computation of the peak detection and alignment steps for larger studies. This pipeline has been successfully evaluated for studies ranging from 200 to 1600 samples. By specifically isolating high quality and reliable signals pertaining to carbon-containing compounds in untargeted LC/HRMS data sets from larger studies, IDSL.IPA opens new opportunities for discovering new biological insights in the population-scale metabolomics and exposomics projects. The package is available in the R CRAN repository at https://cran.r-project.org/package=IDSL.IPA.

9-cis-ß-Apo-10'-carotenal is the precursor of strigolactones in planta.

MAIN CONCLUSION: 13C-isotope feeding experiments demonstrate that the apocarotenoid 9-cis-ß-apo-10'-carotenal is the precursor of several strigolactones in rice, providing a direct, in planta evidence for its role in strigolactone biosynthesis. Strigolactones (SLs) are plant hormone that regulates plant architecture and mediates rhizospheric communications. Previous in vitro studies using heterogously produced enzymes unraveled the conversion of all-trans-ß-carotene via the intermediate 9-cis-ß-apo-10'-carotenal into the SL precursor carlactone. However, a direct evidence for the formation of SLs from 9-cis-ß-apo-10'-carotenal is still missing. To provide this evidence, we supplied rice seedlings with 13C-labeled 9-cis-ß-apo-10'-carotenal and analyzed their SLs by LC-MS. Our results show that 9-cis-ß-apo-10'-carotenal is the SL precursor in planta and reveal, for the first time, the application of labeled long-chain apocarotenoids as a promising approach to investigate apocarotenoid metabolism and the genesis of carotenoid-derived growth regulators and signaling molecules.

Metabolic flux analysis of the non-transitory starch tradeoff for lipid production in mature tobacco leaves.

The metabolic plasticity of tobacco leaves has been demonstrated via the generation of transgenic plants that can accumulate over 30% dry weight as triacylglycerols. In investigating the changes in carbon partitioning in these high lipid-producing (HLP) leaves, foliar lipids accumulated stepwise over development. Interestingly, non-transient starch was observed to accumulate with plant age in WT but not HLP leaves, with a drop in foliar starch concurrent with an increase in lipid content. The metabolic carbon tradeoff between starch and lipid was studied using 13CO2-labeling experiments and isotopically nonstationary metabolic flux analysis, not previously applied to the mature leaves of a crop. Fatty acid synthesis was investigated through assessment of acyl-acyl carrier proteins using a recently derived quantification method that was extended to accommodate isotopic labeling. Analysis of labeling patterns and flux modeling indicated the continued production of unlabeled starch, sucrose cycling, and a significant contribution of NADP-malic enzyme to plastidic pyruvate production for the production of lipids in HLP leaves, with the latter verified by enzyme activity assays. The results suggest an inherent capacity for a developmentally regulated carbon sink in tobacco leaves and may in part explain the uniquely successful leaf lipid engineering efforts in this crop.

Genome-based selection and application of food-grade microbes for chickpea milk fermentation towards increased L-lysine content, elimination of indigestible sugars, and improved flavour.

BACKGROUND: Plant-based milk alternatives are more popular than ever, and chickpea-based milks are among the most commercially relevant products. Unfortunately, limited nutritional value because of low levels of the essential amino acid L-lysine, low digestibility and unpleasant taste are challenges that must be addressed to improve product quality and meet consumer expectations. RESULTS: Using in-silico screening and food safety classifications, 31 strains were selected as potential L-lysine producers from approximately 2,500 potential candidates. Beneficially, 30% of the isolates significantly accumulated amino acids (up to 1.4 mM) during chickpea milk fermentation, increasing the natural level by up to 43%. The best-performing strains, B. amyloliquefaciens NCC 156 and L. paracasei subsp. paracasei NCC 2511, were tested further. De novo lysine biosynthesis was demonstrated in both strains by 13C metabolic pathway analysis. Spiking small amounts of citrate into the fermentation significantly activated L-lysine biosynthesis in NCC 156 and stimulated growth. Both microbes revealed additional benefits in eliminating indigestible sugars such as stachyose and raffinose and converting off-flavour aldehydes into the corresponding alcohols and acids with fruity and sweet notes. CONCLUSIONS: B. amyloliquefaciens NCC 156 and L. paracasei subsp. paracasei NCC 2511 emerged as multi-benefit microbes for chickpea milk fermentation with strong potential for industrial processing of the plant material. Given the high number of L-lysine-producing isolates identified in silico, this concept appears promising to support strain selection for food fermentation.

A precise and rapid isotopomic analysis of small quantities of cholesterol at natural abundance by optimized 1H-13C 2D NMR.

Cholesterol, the principal zoosterol, is a key metabolite linked to several health complications. Studies have shown its potential as a metabolic biomarker for predicting various diseases and determining food origin. However, the existing INEPT (insensitive nuclei enhanced by polarization transfer) 13C position-specific isotope analysis method of cholesterol by NMR was not suitable for very precise analysis of small quantities due to its long acquisition time and therefore is restricted to products rich in cholesterol. In this work, a symmetric and adiabatic heteronuclear single quantum coherence (HSQC) 2D NMR sequence was developed for the high-precision (few permil) analysis of small quantities of cholesterol. Adiabatic pulses were incremented for improving precision and sensitivity. Moreover, several strategies such as the use of non-uniform sampling, linear prediction, and variable recycling time were optimized to reduce the acquisition time. The number of increments and spectral range were also adjusted. The method was developed on a system with a cryogenically cooled probe and was not tested on a room-temperature system. Our new approach allowed analyzing as low as 5 mg of cholesterol in 31 min with a long-term repeatability lower than 2‰ on the 24 non-quaternary carbon atoms of the molecule comparing to 16.2 h for the same quantity using the existing INEPT method. This result makes conceivable the isotope analysis of matrices low in cholesterol. Graphical abstract.

Anaerobic oxidation of methane coupled to sulfate reduction: Consortium characteristics and application in co-removal of H2S and methane.

Anaerobic sludge from a sewage treatment plant was used to acclimatize microbial colonies capable of anaerobic oxidation of methane (AOM) coupled to sulfate reduction. Clone libraries and fluorescence in situ hybridization were used to investigate the microbial population. Sulfate-reducing bacteria (SRB) (e.g., Desulfotomaculum arcticum and Desulfobulbus propionicus) and anaerobic methanotrophic archaea (ANME) (e.g., Methanosaeta sp. and Methanolinea sp.) coexisted in the enrichment. The archaeal and bacterial cells were randomly or evenly distributed throughout the consortia. Accompanied by sulfate reduction, methane was oxidized anaerobically by the consortia of methane-oxidizing archaea and SRB. Moreover, CH4 and SO42- were consumed by methanotrophs and sulfate reducers with CO2 and H2S as products. The H3CSH produced by methanotrophy was an intermediate product during the process. The methanotrophic enrichment was inoculated in a down-flow biofilter for the treatment of methane and H2S from a landfill site. On average, 93.33% of H2S and 10.71% of methane was successfully reduced in the biofilter. This study tries to provide effective method for the synergistic treatment of waste gas containing sulfur compounds and CH4.

Microbial mechanisms of carbon priming effects revealed during the interaction of crop residue and nutrient inputs in contrasting soils.

Agronomic practices such as crop residue return and additional nutrient supply are recommended to increase soil organic carbon (SOC) in arable farmlands. However, changes in the priming effect (PE) on native SOC mineralization in response to integrated inputs of residue and nutrients are not fully known. This knowledge gap along with a lack of understanding of microbial mechanisms hinders the ability to constrain models and to reduce the uncertainty to predict carbon (C) sequestration potential. Using a 13 C-labeled wheat residue, this 126-day incubation study examined the dominant microbial mechanisms that underpin the PE response to inputs of wheat residue and nutrients (nitrogen, phosphorus and sulfur) in two contrasting soils. The residue input caused positive PE through "co-metabolism," supported by increased microbial biomass, C and nitrogen (N) extracellular enzyme activities (EEAs), and gene abundance of certain microbial taxa (Eubacteria, ß-Proteobacteria, Acidobacteria, and Fungi). The residue input could have induced nutrient limitation, causing an increase in the PE via "microbial nutrient mining" of native soil organic matter, as suggested by the low C-to-nutrient stoichiometry of EEAs. At the high residue, exogenous nutrient supply (cf. no-nutrient) initially decreased positive PE by alleviating nutrient mining, which was supported by the low gene abundance of Eubacteria and Fungi. However, after an initial decrease in PE at the high residue with nutrients, the PE increased to the same magnitude as without nutrients over time. This suggests the dominance of "microbial stoichiometry decomposition," supported by higher microbial biomass and EEAs, while Eubacteria and Fungi increased over time, at the high residue with nutrients cf. no-nutrient in both soils. Our study provides novel evidence that different microbial mechanisms operate simultaneously depending on organic C and nutrient availability in a residue-amended soil. Our results have consequences for SOC modeling and integrated nutrient management employed to increase SOC in arable farmlands.

Metabolism of fructose in B-cells: A 13C NMR spectroscopy based stable isotope tracer study.

Earlier studies on glucose metabolism in B-cells suggested an active TCA cycle in both naïve B cells and differentiated IgA plasma cells. Glycolysis was shown to be more active in IgA plasma cells than naïve B-cells. There have been no reports on the metabolism of fructose in B-cells. Fructose is a major sugar present in the western diet. Thus, we have investigated the metabolism of fructose in B-cells including the effect of glucose on the metabolism of fructose. In this study, using 13C NMR spectroscopy and [U-13C]fructose and [U-13C]glucose as stable 13C isotope tracers, we investigated the metabolic fate of fructose and glucose in B-cells. B-cells showed mitochondrial oxidation of fructose when administered alone, but showed diminished oxidation of fructose in the presence of glucose. On the other hand, fructose did not significantly affect the mitochondrial metabolism of glucose.

Incomplete Wood-Ljungdahl pathway facilitates one-carbon metabolism in organohalide-respiring Dehalococcoides mccartyi.

The acetyl-CoA "Wood-Ljungdahl" pathway couples the folate-mediated one-carbon (C1) metabolism to either CO2 reduction or acetate oxidation via acetyl-CoA. This pathway is distributed in diverse anaerobes and is used for both energy conservation and assimilation of C1 compounds. Genome annotations for all sequenced strains of Dehalococcoides mccartyi, an important bacterium involved in the bioremediation of chlorinated solvents, reveal homologous genes encoding an incomplete Wood-Ljungdahl pathway. Because this pathway lacks key enzymes for both C1 metabolism and CO2 reduction, its cellular functions remain elusive. Here we used D. mccartyi strain 195 as a model organism to investigate the metabolic function of this pathway and its impacts on the growth of strain 195. Surprisingly, this pathway cleaves acetyl-CoA to donate a methyl group for production of methyl-tetrahydrofolate (CH3-THF) for methionine biosynthesis, representing an unconventional strategy for generating CH3-THF in organisms without methylene-tetrahydrofolate reductase. Carbon monoxide (CO) was found to accumulate as an obligate by-product from the acetyl-CoA cleavage because of the lack of a CO dehydrogenase in strain 195. CO accumulation inhibits the sustainable growth and dechlorination of strain 195 maintained in pure cultures, but can be prevented by CO-metabolizing anaerobes that coexist with D. mccartyi, resulting in an unusual syntrophic association. We also found that this pathway incorporates exogenous formate to support serine biosynthesis. This study of the incomplete Wood-Ljungdahl pathway in D. mccartyi indicates a unique bacterial C1 metabolism that is critical for D. mccartyi growth and interactions in dechlorinating communities and may play a role in other anaerobic communities.

Monitoring CO2 emissions to gain a dynamic view of carbon allocation to arbuscular mycorrhizal fungi.

Quantification of carbon (C) fluxes in mycorrhizal plants is one of the important yet little explored tasks of mycorrhizal physiology and ecology. 13CO2 pulse-chase labelling experiments are increasingly being used to track the fate of C in these plant-microbial symbioses. Nevertheless, continuous monitoring of both the below- and aboveground CO2 emissions remains a challenge, although it is necessary to establish the full C budget of mycorrhizal plants. Here, a novel CO2 collection system is presented which allows assessment of gaseous CO2 emissions (including isotopic composition of their C) from both belowground and shoot compartments. This system then is used to quantify the allocation of recently fixed C in mycorrhizal versus nonmycorrhizal Medicago truncatula plants with comparable biomass and mineral nutrition. Using this system, we confirmed substantially greater belowground C drain in mycorrhizal versus nonmycorrhizal plants, with the belowground CO2 emissions showing large variation because of fluctuating environmental conditions in the glasshouse. Based on the assembled 13C budget, the C allocation to the mycorrhizal fungus was between 2.3% (increased 13C allocation to mycorrhizal substrate) and 2.9% (reduction of 13C allocation to mycorrhizal shoots) of the plant gross photosynthetic production. Although the C allocation to shoot respiration (measured during one night only) did not differ between the mycorrhizal and nonmycorrhizal plants under our experimental conditions, it presented a substantial part (∼10%) of the plant C budget, comparable to the amount of CO2 released belowground. These results advocate quantification of both above- and belowground CO2 emissions in future studies.

Sparse (13)C labelling for solid-state NMR studies of P. pastoris expressed eukaryotic seven-transmembrane proteins.

We demonstrate a novel sparse (13)C labelling approach for methylotrophic yeast P. pastoris expression system, towards solid-state NMR studies of eukaryotic membrane proteins. The labelling scheme was achieved by co-utilizing natural abundance methanol and specifically (13)C labelled glycerol as carbon sources in the expression medium. This strategy improves the spectral resolution by 1.5 fold, displays site-specific labelling patterns, and has advantages for collecting long-range distance restraints for structure determination of large eukaryotic membrane proteins by solid-state NMR.

The anticonvulsant actions of carisbamate associate with alterations in astrocyte glutamine metabolism in the lithium-pilocarpine epilepsy model.

As reported previously, in the lithium-pilocarpine model of temporal lobe epilepsy (TLE), carisbamate (CRS) produces strong neuroprotection, leads to milder absence-like seizures, and prevents behavioral impairments in a subpopulation of rats. To understand the metabolic basis of these effects, here we injected 90 mg/kg CRS or vehicle twice daily for 7 days starting 1 h after status epilepticus (SE) induction in rats. Two months later, we injected [1-13 C]glucose and [1,2-13 C]acetate followed by head microwave fixation after 15 min. 13 C incorporation into metabolites was analyzed using 13 C magnetic resonance spectroscopy. We found that SE reduced neuronal mitochondrial metabolism in the absence but not in the presence of CRS. Reduction in glutamate level was prevented by CRS and aspartate levels were similar to controls only in rats displaying absence-like seizures after treatment [CRS-absence-like epilepsy (ALE)]. Glutamine levels in CRS-ALE rats were higher compared to controls in hippocampal formation and limbic structures while unchanged in rats displaying motor spontaneous recurrent seizures after treatment (CRS-TLE). Astrocytic mitochondrial metabolism was reduced in CRS-TLE, and either enhanced or unaffected in CRS-ALE rats, which did not affect the transfer of glutamine from astrocytes to neurons. In conclusion, CRS prevents reduction in neuronal mitochondrial metabolism but its effect on astrocytes is likely key in determining outcome of treatment in this model. To understand the metabolic basis of the strong neuroprotection and reduction in seizure severity caused by carisbamate (CRS) in the lithium-pilocarpine (Li-Pilo) model of temporal lobe epilepsy (TLE), we injected CRS for 7 days starting 1 h after status epilepticus and 2 months later [1-13 C]glucose and [1,2-13 C]acetate. 13 C Magnetic resonance spectroscopy analysis was performed on brain extracts and we found that CRS prevented reduction in neuronal mitochondrial metabolism but its effect on astrocytes was likely key in determining outcome of treatment in this model. ALE = absence like epilepsy; acetyl CoA = acetyl coenzyme A; GS = glutamine synthetase; PAG = phosphate activated glutaminase; PC = pyruvate carboxylase; OAA = oxaloacetate; TCA cycle = tricarboxylic acid cycle.

Development and validation of a dynamic range-extended LC-MS/MS multi-analyte method for 11 different postmortem matrices for redistribution studies applying solvent calibration and additional (13)C isotope monitoring.

Postmortem redistribution (PMR) is one of numerous problems in postmortem toxicology making correct interpretation of measured drug concentrations difficult or even impossible. Time-dependent PMR in peripheral blood and especially in tissue samples is still under-explored. For further investigation, an easy applicable method for the simultaneous quantitation of over 80 forensically relevant compounds in 11 different postmortem matrices should be developed and validated overcoming the challenges of high inter-matrix and intra-matrix concentration variances. Biopsy samples (20 mg) or body fluids (20 µL) were spiked with an analyte mix and deuterated internal standards, extracted by liquid-liquid extraction, and analyzed by liquid chromatography tandem mass spectrometry (LC-MS/MS). For highest applicability, an easy solvent calibration was used. Furthermore, time-consuming dilution of high concentration samples showing detector saturation was circumvented by two overlapping calibration curves using (12)C isotope monitoring for low concentrations and (13)C isotopes for high concentration, respectively. The method was validated according to international guidelines with modifications. Matrix effects and extraction efficiency were strongly matrix and analyte dependent. In general, brain and adipose tissue produced the highest matrix effects, whereas cerebrospinal fluid showed the least matrix effects. Accuracy and precision results were rather matrix independent with some exceptions. Despite using an external solvent calibration, the accuracy requirements were fulfilled for 66 to 81 % of the 83 analytes. Depending on the matrix, 75-93 % of the analytes showed intra-day precisions at <20 %. (12)C and (13)C calibrations gave comparable results and proved to be a useful tool in expanding the dynamic range.

Triheptanoin partially restores levels of tricarboxylic acid cycle intermediates in the mouse pilocarpine model of epilepsy.

Triheptanoin, the triglyceride of heptanoate, is anticonvulsant in various epilepsy models. It is thought to improve energy metabolism in the epileptic brain by re-filling the tricarboxylic acid (TCA) cycle with C4-intermediates (anaplerosis). Here, we injected mice with [1,2-(13) C]glucose 3.5-4 weeks after pilocarpine-induced status epilepticus (SE) fed either a control or triheptanoin diet. Amounts of metabolites and incorporations of (13) C were determined in extracts of cerebral cortices and hippocampal formation and enzyme activity and mRNA expression were quantified. The percentage enrichment with two (13) C atoms in malate, citrate, succinate, and GABA was reduced in hippocampal formation of control-fed SE compared with control mice. Except for succinate, these reductions were not found in triheptanoin-fed SE mice, indicating that triheptanoin prevented a decrease of TCA cycle capacity. Compared to those on control diet, triheptanoin-fed SE mice showed few changes in most other metabolite levels and their (13) C labeling. Reduced pyruvate carboxylase mRNA and enzyme activity in forebrains and decreased [2,3-(13) C]aspartate amounts in cortex suggest a pyruvate carboxylation independent source of C-4 TCA cycle intermediates. Most likely anaplerosis was kept unchanged by carboxylation of propionyl-CoA derived from heptanoate. Further studies are proposed to fully understand triheptanoin's effects on neuroglial metabolism and interaction.

Strigolactone biosynthesis in rice can occur via a 9-cis-3-OH-10'-apo-ß-carotenal intermediate.

Strigolactones (SLs) play a crucial role in regulating plant architecture and mediating rhizosphere interactions. They are synthesized from all-trans-ß-carotene converted into the intermediate carlactone (CL) via the intermediate 9-cis-ß-apo-10'-carotenal. Recent studies indicate that plants can also synthesize 3-OH-CL from all-trans-ß-zeaxanthin via the intermediate 9-cis-3-OH-ß-apo-10'-carotenal. However, the question of whether plants can form bioactive SLs from 9-cis-3-OH-ß-apo-10'-carotenal remains elusive. In this study, we supplied the 13 C-labeled 9-cis-3-OH-ß-apo-10'-carotenal to rice seedlings and monitored the synthesis of SLs using liquid chromatography-mass spectrometry (LC-MS) and Striga bioassay. We further validated the biological activity of 9-cis-3-OH-ß-apo-10'-carotenal-derived SLs using the ccd7/d17 SL-deficient mutant, which demonstrated increased Striga seed-germinating activity and partial rescue of tiller numbers and plant height. Our results establish 9-cis-3-OH-ß-apo-10'-carotenal as a significant SL biosynthetic intermediate with implications for understanding plant hormonal functions and potential applications in agriculture.

Tracing the path of disruption: 13C isotope applications in traumatic brain injury-induced metabolic dysfunction.

Cerebral metabolic dysfunction is a critical pathological hallmark observed in the aftermath of traumatic brain injury (TBI), as extensively documented in clinical investigations and experimental models. An in-depth understanding of the bioenergetic disturbances that occur following TBI promises to reveal novel therapeutic targets, paving the way for the timely development of interventions to improve patient outcomes. The 13C isotope tracing technique represents a robust methodological advance, harnessing biochemical quantification to delineate the metabolic trajectories of isotopically labeled substrates. This nuanced approach enables real-time mapping of metabolic fluxes, providing a window into the cellular energetic state and elucidating the perturbations in key metabolic circuits. By applying this sophisticated tool, researchers can dissect the complexities of bioenergetic networks within the central nervous system, offering insights into the metabolic derangements specific to TBI pathology. Embraced by both animal studies and clinical research, 13C isotope tracing has bolstered our understanding of TBI-induced metabolic dysregulation. This review synthesizes current applications of isotope tracing and its transformative potential in evaluating and addressing the metabolic sequelae of TBI.

Selective uneven enrichment of soil organic carbon among different-sized sediments under a rain-induced overland flow: 13C stable isotope evidence.

Soil organic carbon (SOC) enrichment varies among sediments of different sizes during rain-induced overland flow erosion. This selective transport of SOC is complex in conjunction with the exposure of labile and stable organic carbon (OC), accompanied by heterogeneous aggregate disintegration under raindrop effects. Utilizing the variations in δ13C values of SOC fractions, we traced this selective transport, linking it to aggregate-wrapped SOC changes during erosion. A modified soil pan facilitated the simultaneous monitoring of splash and sheet erosion via artificially simulated rainfall, with control over the intensity and slope. Aggregate composition, SOC distribution, and δ13C values in the erosion samples were analyzed. The results indicated that distinct sorting existed within the aggregate fragments. Along with SOC variation among different sediment sizes, the proportions of clay and fine silt within sediment aggregates increased as a function of slope and rainfall intensity, whereas particulate OC within aggregates decreased. The SOC enrichment ratios (ERocs) and δ13C values in splash-eroded sediments were positively correlated with those in sheet-eroded sediments. The ERocs in splash-eroded sediments were lower than those in sheet-eroded sediments, but δ13C values were the opposite. Moreover, δ13C values of SOC enriched in sediment particles of all sizes from aggregate stripping were lower than those of the original soil. This indicates that raindrop hits promote heavy C loss during sheet erosion, which is different for mineral-associated and particulate OC. As the slope and rainfall intensity increased, δ13C values for all sediment sizes decreased over the course of erosion. Interestingly, the highest δ13C values were observed under a rainfall intensity of 60 mm h-1, whereas the highest SOC concentrations were noted on a 5° slope. These observations suggest divergent mechanisms affect δ13C values and SOC concentrations in eroded sediments. All these results verified that selective sorting existed for the light SOC fraction. Finally, the internal selective transport of one SOC fraction may explain the enhanced mineralization and reaggregation capacity of the deposited sediments.