RESUMEN
Atmospheric methane (CH4) acts as a key contributor to global warming. As CH4 is a short-lived climate forcer (12 years atmospheric lifespan), its mitigation represents the most promising means to address climate change in the short term. Enteric CH4 (the biosynthesized CH4 from the rumen of ruminants) represents 5.1% of total global greenhouse gas (GHG) emissions, 23% of emissions from agriculture, and 27.2% of global CH4 emissions. Therefore, it is imperative to investigate methanogenesis inhibitors and their underlying modes of action. We hereby elucidate the detailed biophysical and thermodynamic interplay between anti-methanogenic molecules and cofactor F430 of methyl coenzyme M reductase and interpret the stoichiometric ratios and binding affinities of sixteen inhibitor molecules. We leverage this as prior in a graph neural network to first functionally cluster these sixteen known inhibitors among ~54,000 bovine metabolites. We subsequently demonstrate a protocol to identify precursors to and putative inhibitors for methanogenesis, based on Tanimoto chemical similarity and membrane permeability predictions. This work lays the foundation for computational and de novo design of inhibitor molecules that retain/ reject one or more biochemical properties of known inhibitors discussed in this study.
RESUMEN
Gut microbiota has been implicated in many disorders including Autism Spectrum Disorder (ASD). ASD is a neurodevelopmental brain disorder affecting individuals leading to restricted and repetitive pattern of behaviour and disruption of communication and social interactions. Altered microbiome and the presence or absence of key species capable of affecting specific responses in levels of their fermentation products are reflected in the urinary metabolite profile of patients. The aim of our study is to develop an improvised method for the detection and quantification of urinary p-cresol levels which could serve as an indicator for GI microbial dysbiosis. The p-cresol analysis was achieved using HPLC by a reverse phase C18 column with mobile phase composition of Acetonitrile/water/formic acid (10:90:0.05, v/v/v) in an isocratic mode of elution with a flow rate of 1.0 mL/min. The mass analysis of p-cresol was performed using LC-MS [Triple Quadrupole Liquid Chromatography Mass Spectrometer] in negative ESI mode with electron multiplier detector. p-cresol was eluted at a retention time of approximately 3.4 min. The standard calibration curves had a superior regression coefficient of greater than 0.99 (R2 > 0.99) and were linear over a range from 0.0005 mg/mL to 0.015 mg/mL. The method was validated by analysis of six replicates with 0.08% relative standard deviation and method detection and quantification limits were 20 ng/mL and 50 ng/mL respectively. Further validation of method on real urine samples from two groups of children (Control population:< 10 years of age; 5M: 3F and ASD individuals: <10 years of age; All males) showed that detection was effective over a wide range of metabolite at levels as high as 149.73 µg/mL to as low as 0.897 µg/mL. This study reports a rapid, validated and sensitive method for the detection of p-cresol in urine samples.