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Biomarkers for diagnosing the retained placenta in cows-does the analysis of serum proteome keep the secret?

ABSTRACT: Retained placenta (RP), a quite common disorder in dairy cows, shows a high negative impact on their health status and milk production. Therefore, the biomarkers indicating the risk for RP could yield benefits in dairy management. The study aimed to investigate the serum proteome as the source of the biomarkers for diagnosing the RP. Using the high-resolution liquid chromatography-tandem mass spectrometry, we compared the composition of serum proteome between 9 cows with the RP and 6 with the physiologic puerperium (PP). Among 651 identified proteins, seven had higher, and three showed lower abundance in RP than in the PP group. The differently abundant proteins participated in 15 pathways: six related to hemostasis, three involved in the lipoprotein metabolism, and the remaining ones associated with redox homeostasis, post-translational modification, scavenging, etc. Validation of the proteomic results, using ELISA and spectrophotometric assays, showed that the two identified proteins, haptoglobin, and lipopolysaccharide-binding protein, could represent reliable RP biomarkers. Our results confirmed that the analysis of serum proteome shed "the light" on the counterparty piece(s) of the RP pathogenesis "puzzle" and could be an efficient "tool" in mining for RP biomarkers.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Bos Taurus (bovine)

TISSUE(S): Blood Serum

SUBMITTER: Josipa Kuleš

LAB HEAD: Vladimir Mrljak

PROVIDER: PXD034963 | Pride | 2023-01-13

REPOSITORIES: Pride

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Project description:Post-partum uterine inflammation (endometritis) is associated with lower fertility at both the time of infection and after the inflammation has resolved. It was hypothesized that aberrant DNA methylation may be involved in the sub-fertility associated with post-partum uterine inflammation. The objective of this study was to characterize genome-wide DNA methylation and gene expression in the endometrium of dairy cows with sub-clinical endometritis. Endometrial tissues were obtained at 29 days post-partum (n=12) and Agilent two-colour microarrays were used to characterize transcription and DNA methylation profiles. Analyses revealed 1,856 probes to be differentially expressed in animals with subclinical endometritis (SUI, n=6) compared with control cows (NUI, n=6, P<0.05, Storey Multiple testing correction). No significant associations among DNA methylation and gene expression were detected. Further analysis of gene expression data using GeneGo Metacore and Gene Set Enrichment Analysis identified several pathways and processes enriched in the comparison. Several pathways that are involved in the innate immune response were enriched in SUI cows. Consistent with the presence of microorganisms in the uterus, there was enrichment for the Toll-like receptor (TLR) signaling pathway, including increased expression of the transcription factor NFKB1, the pro-inflammatory cytokines IL1A and IL1B, downstream chemokines, cytokines, and acute phase and antimicrobial proteins in the endometrium of SUI cows. Furthermore, the chemokine signaling pathway was enriched in SUI cows, with increased expression of genes that attract cells of the innate immune system. Increased expression of IL-8 and CXCL6, chemotactic factors for recruitment of neutrophils along with the immune cell surface marker PTPRC in SUI cows is consistent with the greater number of polymorphonuclear cells present in the uterus of these cows. Several antimicrobial peptides (LAP, TAP, DEFB1, DEFB10, DEFB103B, DEFB7) and acute phase proteins, including SAA3, LBP, and the complement gene CFB, had greater expression in SUI cows. Gene expression profiles in cows with subclinical endometritis in this study indicate that the immune response is activated, potentially resulting in a local pro-inflammatory environment in the uterus. If this period of inflammation is prolonged, it could result in tissue damage or failure to complete involution of the uterus, which may create a sub-optimal environment for future pregnancy. Agilent two-colour microarrays were used to characterize DNA methylation profiles in cows with subclinical endometritis (SUI, n=6) compared to control cows (NUI, n=6). Endometrial tissues (caruncular, intercaruncular) were obtained at 29 days post-partum.

			Action	DRS
	checksum.txt	Txt
	cow1-_2__PeptideGroups.txt	Txt
	cow1-_2__ProteinGroups.txt	Txt
	cow1-_2__Proteins.txt	Txt
	cow1.mgf	Mgf

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Biomarkers for diagnosing the retained placenta in cows-does the analysis of serum proteome keep the secret?

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