Uterine Fluid Extracellular Vesicles Proteome Is Altered During the Estrous Cycle

Piibor, Johanna; Waldmann, Andres; Dissanayake, Keerthie; Andronowska, Aneta; Ivask, Marilin; Prasadani, Madhusha; Kavak, Ants; Kodithuwakku, Suranga; Fazeli, Alireza

doi:10.1016/j.mcpro.2023.100642

Cited by 5 publications

(12 citation statements)

References 67 publications

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“…The UF-EV enrichment was performed as described previously [ 43 ]. The samples were concentrated at 4000 g in 4 °C using Amicon ® Ultra 15 mL centrifugal filters (10 kDa cut-off, Merck Millipore Ltd., Darmstadt, Germany) to 500 µL.…”

Section: Methodsmentioning

confidence: 99%

“…The characterization of UF-EVs was performed following previously published protocol [ 43 ] in accordance with the International Society for Extracellular Vesicles guidelines [ 44 ]. Briefly, particle concentration and size were measured with an NTA ZetaView ® device (PMX 110 V3.0 instrument by Particle Metrix GmbH, Inning am Ammersee, Germany) coupled with ZetaView NTA software (v3.0).…”

Section: Methodsmentioning

confidence: 99%

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Investigation of Uterine Fluid Extracellular Vesicles’ Proteomic Profiles Provides Novel Diagnostic Biomarkers of Bovine Endometritis

Piibor,

Waldmann,

Prasadani

et al. 2024

Biomolecules

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Cow uterine infections pose a challenge in dairy farming, resulting in reproductive disorders. Uterine fluid extracellular vesicles (UF-EVs) play a key role in cell-to-cell communication in the uterus, potentially holding the signs of aetiology for endometritis. We used mass spectrometry-based quantitative shotgun proteomics to compare UF-EV proteomic profiles in healthy cows (H), cows with subclinical (SE) or clinical endometritis (CLE) sampled at 28–35 days postpartum. Functional analysis was performed on embryo cultures with the exposure to different EV types. A total of 248 UF-EV proteins exhibited differential enrichment between the groups. Interestingly, in SE, EV protein signature suggests a slight suppression of inflammatory response compared to CLE-UF-EVs, clustering closer with healthy cows’ profile. Furthermore, CLE-UF-EVs proteomic profile highlighted pathways associated with cell apoptosis and active inflammation aimed at pathogen elimination. In SE-UF-EVs, the regulation of normal physiological status was aberrant, showing cell damage and endometrial repair at the same time. Serine peptidase HtrA1 (HTRA1) emerged as a potential biomarker for SE. Supplementation of CLE- and SE-derived UF-EVs reduced the embryo developmental rates and quality. Therefore, further research is warranted to elucidate the precise aetiology of SE in cattle, and HTRA1 should be further explored as a potential diagnostic biomarker.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Investigation of Uterine Fluid Extracellular Vesicles’ Proteomic Profiles Provides Novel Diagnostic Biomarkers of Bovine Endometritis

Piibor,

Waldmann,

Prasadani

et al. 2024

Biomolecules

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“…The standards were suspended in particle-free Milli-Q water, whereas the EV samples were diluted in PBS for analyses. The particle size distribution and number were counted at 11 frames per cycle, under a sensitivity of 72 and a shutter value of 100 [32].…”

Section: Nanoparticle Tracking Analysis (Nta)mentioning

confidence: 99%

“…Western blot analysis was performed on the milk EVs enriched with TFF and whey sample, according to the method described elsewhere [32]. Briefly, EVs were lysed and proteins were extracted using RIPA buffer (Thermo Fisher Scientific, Waltham, MA, USA), with a protease inhibitor cocktail (cat.…”

Section: Sds-page and Western Blottingmentioning

confidence: 99%

Harnessing Nature’s Defence: The Antimicrobial Efficacy of Pasteurised Cattle Milk-Derived Extracellular Vesicles on Staphylococcus aureus ATCC 25923

Sapugahawatte,

Godakumara,

Mäesaar

et al. 2024

IJMS

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Increasing antimicrobial resistance (AMR) challenges conventional antibiotics, prompting the search for alternatives. Extracellular vesicles (EVs) from pasteurised cattle milk offer promise, due to their unique properties. This study investigates their efficacy against five pathogenic bacteria, including Staphylococcus aureus ATCC 25923, aiming to combat AMR and to develop new therapies. EVs were characterised and tested using various methods. Co-culture experiments with S. aureus showed significant growth inhibition, with colony-forming units decreasing from 2.4 × 105 CFU/mL (single dose) to 7.4 × 104 CFU/mL (triple doses) after 12 h. Milk EVs extended lag time (6 to 9 h) and increased generation time (2.8 to 4.8 h) dose-dependently, compared to controls. In conclusion, milk EVs exhibit dose-dependent inhibition against S. aureus, prolonging lag and generation times. Despite limitations, this suggests their potential in addressing AMR.

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“…Exosomes have become a focal point in biomedical research due to their crucial influence on various biological functions and their ability to modulate microenvironments 21,22 . Exosomes, classified as membrane-bound nanovesicles with sizes ranging from 30 to 200 nm 23 , are released from diverse cell types and bodily fluids, including blood 24 , saliva 25 , breast milk 26 , urine 27 , tears 28 , and vaginal discharge 29 . These small vesicles transport bioactive compounds like proteins, nucleic acids, and lipids 30,31 .…”

Section: Mainmentioning

confidence: 99%

Exosomal miRNAs from maternal vaginal discharge as biomarkers for preterm labor: non-invasive liquid biopsy

Kim,

Park,

Lee

et al. 2024

Preprint

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Preterm labor is a serious issue that can lead to preterm birth, posing significant risks to both the mother and the neonate. Despite the high incidence of approximately 15 million preterm births worldwide per year, there is a lack of sufficient strategies for predicting and preventing preterm labor. Here, we found that exosomal miRNAs in maternal vaginal discharge can serve as biomarkers for early diagnosis of life-threatening conditions in both the mother and neonate. Our non-invasive biopsy of vaginal discharge using a swab allows us to isolate enriched exosomes via an advanced microfluidic platform called BEST (Biologically intact Exosome Separation Technology). We have identified specific miRNAs differentially expressed in mothers with preterm labor compared to those with full-term mothers. These miRNAs included hsa-miR-206, which was up-regulated in preterm labor, and hsa-miR-3674, hsa-miR-365a-5p, and hsa-miR-193b-3p, which were down-regulated. We believe our discovery of unique miRNAs as biomarkers can aid in early detection and effective treatment of preterm labor, potentially revolutionizing global healthcare.

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Uterine Fluid Extracellular Vesicles Proteome Is Altered During the Estrous Cycle

Cited by 5 publications

References 67 publications

Investigation of Uterine Fluid Extracellular Vesicles’ Proteomic Profiles Provides Novel Diagnostic Biomarkers of Bovine Endometritis

Investigation of Uterine Fluid Extracellular Vesicles’ Proteomic Profiles Provides Novel Diagnostic Biomarkers of Bovine Endometritis

Harnessing Nature’s Defence: The Antimicrobial Efficacy of Pasteurised Cattle Milk-Derived Extracellular Vesicles on Staphylococcus aureus ATCC 25923

Exosomal miRNAs from maternal vaginal discharge as biomarkers for preterm labor: non-invasive liquid biopsy

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