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Microbiota biobanking using ice recrystalization inhibitor

ABSTRACT: This study use different ice recrystalization inhibitors (IRIs) for better storing live microbiota. We evaluated whether the addition of IRIs can improve the cultivibility of microbiome and maintain their resposnes to prebiotic kestose. Frozen or fresh microbiota were cultured with or without kestose for 24 hours, and microbiota samples were collected for metaproteomics analysis.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Human Feces Metagenome

SUBMITTER: Xu Zhang

LAB HEAD: Xu Zhang

PROVIDER: PXD034979 | Pride | 2023-02-07

REPOSITORIES: Pride

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An economic and robust TMT labeling approach for high throughput proteomic and metaproteomic analysis.

Creskey Marybeth M Li Leyuan L Ning Zhibin Z Fekete Emily Ef EE Mayne Janice J Walker Krystal K Ampaw Anna A Ben Robert R Zhang Xu X Figeys Daniel D

Proteomics 20230113 21-22

Multiplexed quantitative proteomics using tandem mass tag (TMT) is increasingly used in -omic study of complex samples. While TMT-based proteomics has the advantages of the higher quantitative accuracy, fewer missing values, and reduced instrument analysis time, it is limited by the additional reagent cost. In addition, current TMT labeling workflows involve repeated small volume pipetting of reagents in volatile solvents, which may increase the sample-to-sample variations and is not readily sui ...[more]

PMID: 36528842

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Effects of 5 drugs on the human gut microbiome.

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Project description:We systematically investigated the responses of five human gut microbiomes to 21 common sweeteners, using an approach combining high-throughput ex vivo microbiome culturing and metaproteomics to quantify functional changes in different taxa. Hierarchical clustering based on metaproteomic responses of individual microbiomes resulted in two clusters. The first cluster was composed of non-caloric artificial sweeteners (NAS) and two sugar alcohols with shorter carbon backbones (4-5 carbon atoms), and the second cluster was composed of sugar alcohols with longer carbon backbones. The metaproteomic functional responses of the second cluster were similar to the prebiotic fructooligosaccharides and kestose, indicating that these sugar alcohol-type sweeteners have potential prebiotic functions.

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Effect of Edurans secretome of human microbiome

Project description:A rapid ex vivo microbiome assay and metaproteomics approach was used for evaluating the impacts of E. durans derived secretome on a human gut microbiome cultured in vitro.

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Project description:The link between human gut microbiota (a complex group of microorganisms including not only bacteria but also fungi, viruses, etc.,) and the physiological state is nowadays unquestionable. Metaproteomic has emerged as a useful technique to characterize this microbial community, not just taxonomically, but also focusing on specific biological processes carried out by gut microbiota that may have an effect in the host health or pathological state. Cystic fibrosis is a genetic disease in which the microbiota of the respiratory tract determines the patient's survival and differences in composition of gut microbiota of cystic fibrosis patients respect to healthy infants have been reported. In order to characterize this host-microbiota inter-relation, we carried out the metaproteomic study of 30 stool samples from infants with cystic fibrosis.

2022-10-11 | PXD029284 | Pride

Study of human gut microbiota from healthy adults

Project description:The link between the gut microbiota of a human being (a complex group of microorganism including not only bacteria but also fungi, viruses, etc.,) that form an ecosystem in his gastrointestinal tract and his physiological state is nowadays unquestionable. Metaproteomics has emerged as a useful technique to characterize this microbial community, not just taxonomically, but also focusing on specific biological processes carried out by gut microbiota that may have an effect in the host health or pathological state. In order to characterize this host-microbiota inter-relation, we carried out the metaproteomic study of 6 stool samples from 6 healthy adults. A total of 37 080 peptide sequences and 10 686 protein groups were identified in this study. Regarding taxonomic information, we found a total of 247 taxa among 105 were species. Interesting contributions of microbiota metabolism to human host physiology has also been described.

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Evaluation of different protein extraction protocols to study human gut microbiota.

Project description:The link between the gut microbiota and the human physiological state has been demonstrated in recent years. High gut microbiota diversity has been linked to many beneficial functions necessary or human health, while dysbiosis has been correlated to different pathological states. In this context, the study of the gut microbiota results of high relevance been necessary the development of different techniques capable of characterizing this complex ecosystem. Metaproteomics has been proved useful in the characterization of complex protein samples becoming a suitable tool for the study of these microbial communities. However, due to the complexity of these samples, protein extraction protocols may affect metaproteomics results. In this context, we evaluated stool sample processing (SSP) and microbial cell disruption, assessing the impact of different protocol modifications in the number of peptides and proteins identified. We compared different stool processing conditions and microbial cell disruption methods in terms of protein and peptide identifications and taxonomic profiles.

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Deciphering Differential Gut Bacterial Drug Metabolism with Activity-Based Protein Profiling

Project description:Gut bacterial β-glucuronidases (GUS) promote the toxic side effects of therapeutics by reactivating drugs from their inactive glucuronide conjugates. It is increasingly clear that the interindividual variability of bacterial GUS-producing species in the gut microbiota contributes to differential drug responses. Indeed, the anticancer drug irinotecan exhibits variable clinical toxicity outcomes that have been linked to interindividual differences in the composition of the gut microbiota. However, identification of the specific GUS enzymes responsible for drug metabolism in the context of the complexity of the human fecal microbiota has not been achieved. Here we pinpoint the specific bacterial GUS enzymes that reactivate SN-38, the active metabolite of irinotecan, from complex human fecal microbiota samples with activity-based protein profiling (ABPP). We identify and quantify gut bacterial GUS enzymes from human feces with ABPP-enabled proteomics and then integrate this information with ex vivo kinetics to reveal the specific GUS enzymes responsible for the reactivation of SN-38. The same ABPP approach also reveals the molecular basis for differential gut bacterial GUS inhibition between human fecal samples. Taken together, this work provides an unprecedented pipeline to identify the specific bacterial GUS enzymes responsible for drug-induced GI toxicity from the complexity of human feces, which may serve as highly precise biomarkers of clinical outcomes for irinotecan and other therapeutics.

2019-12-03 | PXD014864 | Pride

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