P3044 Characterization of exosomal immune-related micrornas in colostrum and milk from average, low and high immune responder cows
Notice bibliographique
Résumé
The expression of bioactive host defense proteins in bovine colostrum and milk is regulated by miRNAs. MiRNAs are abundant in milk at different lactation stages, often enclosed within exosomes that protect them from degradation and likely allow for cell-to-cell communication. Notably, immune-related miRNA are highly expressed in milk, particularly in colostrum, with potential to regulate gut immunity of the newborn calf and humans. To date, only few studies have evaluated the expression profile of miRNAs in cow’s milk. An interesting observation with these studies was the considerable variation in the total and identified individual miRNAs. These discrepancies were explained by differences in the RNA isolation methods and the background of the cows used in the studies. While high immune responder (HIR) cows have superior colostrum quality and concentrations of bioactive proteins compared with average (A) and low (L) responders, the expression profiles of colostrum and milk miRNAs remain to be elucidated. The central hypothesis of this study is that colostrum and milk-derived exosomes containing immune-related miRNAs are differentially expressed in A and L responders compared with HIR. The objectives of the study are to: (1) establish a reliable protocol for the fractionation of colostrum and milk for further purification and isolation of exosomes-containing miRNAs, and (2) compare and validate the expression profile of colostrum and milk exosomal miRNAs from A, L and HIR. Exosomes were isolated by differential ultracentrifugation and their presence was confirmed by immunogold labeling electron microscopy. Six RNA extraction methods were compared and RNA quantity and quality were assessed by Qubit Fluorometer and bioanalyzer. The total exosomes RNA/protein isolation kit (Invitrogen) had high RNA yield and the least ribosomal contamination. Milk samples from average responders that passed the library generation quality control (four out of 10 samples) were subjected to next-generation sequencing using the Illumina mirRNA-HiSeq2500 and bioinformatics analysis. A set of identified and novel miRNAs is being selected for validation by PCR array. This study provides platform for optimal colostrum and milk exosomal miRNAs identification that is necessary for down-stream applications. These include, but are not limited to, the discovery of novel miRNAs that have the potential to be utilized as biomarkers for milk quality control or for better understanding of the molecular mechanisms that underlie the regulatory role of miRNAs in calves and humans with emphasis on the health benefit of milk.
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Comment cette classification a été obtenuedéplier
Prédiction distillée sur la base complète
Imitation des enseignantsNi prévalence calibrée, ni vérité terrain. Validation humaine à venir. Apprise à partir de 10 348 étiquettes directes de Codex et de 10 348 étiquettes directes de Gemma. Le mode candidate est l'union des têtes enseignantes seuillées; le consensus est leur intersection. Ces sorties portent le statut machine_predicted_unvalidated et ne sont ni des étiquettes humaines ni des étiquettes directes de modèles de pointe.
Scores Codex et Gemma par catégorie
| Catégorie | Codex | Gemma |
|---|---|---|
| Métarecherche | 0,001 | 0,000 |
| Méta-épidémiologie (sens strict) | 0,000 | 0,000 |
| Méta-épidémiologie (sens large) | 0,000 | 0,000 |
| Bibliométrie | 0,000 | 0,000 |
| Études des sciences et des technologies | 0,000 | 0,001 |
| Communication savante | 0,000 | 0,000 |
| Science ouverte | 0,000 | 0,000 |
| Intégrité de la recherche | 0,000 | 0,000 |
| Charge utile insuffisante (le modèle a refusé de juger) | 0,000 | 0,000 |
Scores machine (provisoires)
Les deux têtes enseignantes du modèle étudiant, lues sur ce travail. Un score ordonne la base pour la relecture; il n'affirme jamais une catégorie, et le statut de validation accompagne chaque rangée tel quel.
Scores de référence d'un modèle non mature (critères de maturité non atteints, 7 itérations). Un score ordonne; il n'affirme jamais une catégorie.
score_only:v0-immature-baseline · tel quel depuis la passe de notation : score_only signifie que le nombre peut ordonner les travaux, et qu'aucune étiquette de catégorie n'en découleClassification
machine, non validéePrédiction automatique; un appel candidat d’une seule tête enseignante, pas un consensus.
Le détail, modèle par modèle et score par score, se trouve en fin de page sous « Comment cette classification a été obtenue ».