MALDI-MS-specific signature peaks accurately distinguished patients with HC-HCC from those with HC-LC and CHC. The results indicate the potential of MALDI-MS and the selected peaks to improve HCC surveillance in patients with viral C cirrhosis and chronic hepatitis C.
Study question Does the analysis of the metabolites of the embryonic culture medium can predict the sex of the embryo? Summary answer The presence and quantity of some metabolites in the culture medium can predict the sex of the human embryos. What is known already Advances in analytical techniques for metabolomics have brought the possibility of better tools for the characterization of molecules. Embryonic metabolism can be used as a good indicator of viability, regardless of the morphology of the blastocysts, since differences were observed in the metabolic activities between the days of embryo development and in the rates of live births. Study design, size, duration 16 patients had their embryos biopsied between the months of January to July 2019 in a human reproduction laboratory. All cases had PGT-A indication and after the biopsy, the embryos were frozen. The culture medium samples were individually prepared for metabolites extraction according to the Bligh and Dyer protocol. Controlled ovarian stimulation and dose adjustments according to the response of each patient. The metabolomics analysis was performed by mass spectrometry. Participants/materials, setting, methods Follicular puncture were performed 35 hours after r-hCG. The eggs were kept in individual culture until the blastocyst stage. The blastocysts biopsy was performed (20). After the culture medium was sent to the 337 metabolites analysis by mass spectrometry. 14 molecules with the highest score on the PLS-Da was submitted to the ROC curves showing the power of metabolic analysis to predict the sex of euploid embryos. Besides, we performed the functional enrichment analysis. Main results and the role of chance After the genetic analysis by PGT-a, we obtain 20 euploid embryos, being 12 female embryos and 08 male embryos. Comparing the quantitative target metabolomic analysis of the 337 metabolites in the embryo culture medium, we observed the Asymmetric dimethylarginine, FAD, Malic Acid, Serotonin, increased in female embryos and Adenosine monophosphate, L-Alanine, L-Arginine, Cysteamine, DL-Dopa, Flavin Mononucleotide, Methionine sulfone, Nicotinic acid, L-Tyrosine, Uracil in male embryos. Through the ROC curve, we can verify AUC = 0.937. This result suggests that the metabolomic analysis of the culture medium is valid to be used as a complement of PGT-A to know embryo sex diagnostic. The functional enrichment analysis shows the Asymmetric dimethylarginine and Malic Sulfone metabolism as the principal function alter by female embryos. Limitations, reasons for caution Small number of samples Wider implications of the findings: Further studies are needed to validate these findings for the diagnostic of sex embryos Trial registration number N/A
Study question Does the metabolomic analysis of the embryonic culture medium predict the embryo aneuploidy? Summary answer The presence and quantity of some metabolites in the culture medium can select euploid embryos for transfer. What is known already Advances in analytical techniques for metabolomics have brought the possibility of better tools for the characterization of molecules. Embryonic metabolism can be used as a good indicator of viability, regardless of the morphology of the blastocysts, since differences were observed in the metabolic activities between the days of embryo development and in the rates of live births. Study design, size, duration 17 patients had their embryos biopsied between January to July 2019 in a human reproduction laboratory. All cases had PGT-A indication and after the biopsy, the embryos were frozen. The culture medium samples were individually prepared for metabolites extraction according to the Bligh and Dyer protocol. Controlled ovarian stimulation and dose adjustments according to the response of each patient. The metabolomics analysis was performed by mass spectrometry. Participants/materials, setting, methods Ovum pick up will be performed 35 hours after r-hCG administration. The embryos were kept in individual 50ul drops until the blastocyst stage. The biopsy was performed in 26 blastocysts. The samples were sent to the 337 metabolites analysis by mass spectrometry. 15 molecules with the highest score on the PLS-Da was submitted the ROC curves to illustrate the power of the metabolic ploidy analysis. Besides, we performed the functional enrichment analysis for each group. Main results and the role of chance After the genetic analysis by PGT-a, 10 aneuploid embryos and 16 euploid embryos were found. Comparing the quantitative target metabolomic analysis of the 337 metabolites in the embryo culture medium, we observed the L-Alanine, Cytosine, Guanosine monophosphate, Homocysteine, Hypoxanthine, and Xanthine hiperrepresented in the aneuploid embryos, and the Citrulline, L-Glutamic acid, Kynurenine, L-Leucine, Methionine, Ornithine, L-Phenylalanine, L-Tyrosine, L-Valine were hiperrepresented in the euploid embryos. Through the ROC curve, we can verify AUC = 0.987. This result suggests that the analysis of euploid embryos through the metabolomic analysis of the culture medium is valid to be used as a noninvasive aneuploid diagnostic. The functional enrichment analysis shows the urea cycle and the glycine and serine metabolism as the principal function alter by aneuploid. Limitations, reasons for caution Small number of samples and not validate sample group. Wider implications of the findings: Further studies are needed to validate these findings for the diagnostic of embryo euploidy. Trial registration number N/A
Study question Can oral contraceptives (OCs) influence the metabolism of endometrial stem cells (EnMSC)? Summary answer Altered metabolomic profiling in culture media may be an effect of OC hormonal properties on EnMSC metabolism. What is known already The reconstitution of the endometrial tissue relies on the presence of EnMSC, which reside in the perivascular space in the endometrium. This process can be affected by OCs by decreasing the secretory potential of the glands. In case of high dose OCs, hyperplasia of endometrial vessels and stroma atrophy of the glands may occur. However, the precise effect of OCs on the production and metabolism of EnMSCs remains unknown. Study design, size, duration This was a prospective study that included samples of menstrual blood from 5 volunteers. The study was conducted for one year and received approval from Ethics in Research Committee. Written informed consent was obtained from all participants. Participants/materials, setting, methods Five women with regular menstrual cycles were included in two groups according to the use of OCs: OC (n = 2) and non-OC (n = 3). Samples were collected by the menstrual cup on the second day of the menstrual cycle. The culture medium of EnMSC was collected from each passage. Quantitative metabolomics was performed by multiple reaction monitoring, followed by liquid chromatography mass spectrometry. Data was analyzed by PLS-DA and T-test. Potential biomarkers were assessed by ROC curve. Main results and the role of chance The cells were characterized as mesenchymal stem cells with a range of 47,4% to 85% of positive markers. From 186 metabolites quantified in the culture media of OC and non-OC groups, 15 metabolites were proposed as of high discrimination between groups by the PLS-DA, which also demonstrated a total groups separation on the statistical model. The ROC curve showed that 4 out of 15 metabolites presented more than 80% of sensitivity. These metabolites are Alanine, Phosphatidylcholine (PC) aa C30:0, Glycine and PC aa C32:2, whose concentrations were higher in the OC group than in non-OC group. The Students’ T-Test analysis confirmed that the metabolites with higher discrimination between groups presented significant p values. These metabolites have been associated with several metabolic processes, including energy production and alteration of molecular pathways related to thrombosis and cancer. Limitations, reasons for caution This study was conducted with a small number of participants and further studies are necessary to confirm the findings. Wider implications of the findings The use of OCs could affect endometrial characteristics that are crucial for reproductive success, such as endometrial receptivity. This study provides a preliminary insight into the EnMSCs response to OCs based on specific metabolite signatures, which may contribute to the development of future new reproductive therapies. Trial registration number Foundation for Coordination of Higher Education Personnel (CAPES – Brazil).
In cattle, uterine luminal fluid (ULF) is the main source of molecules that support embryo development and survival during the peri-implantation period. Overarching hypothesis was that peri-estrus changes in ULF volume through accumulation and resorption mechanisms influence ULF composition during the estrous cycle and early pregnancy. Objectives were (1) to characterize individual temporal and spatial changes in ULF volume, endometrial and luteal vascularity, endometrial and luteal size, and progesterone (P4) concentrations during the peri-estrus period in beef heifers and, (2) associate such changes with the metabolite composition in the ULF, four days after estrus. Fourteen Bos indicus heifers that presented a PGF2α responsive CL received 500 µg PGF2α analog i.m. and were examined daily by rectal B-mode and pulse-wave color-Doppler ultrasonography until the fifth day after estrus (estrus = d 0). Plasma P4 was measured daily. On d 4, the uterine body was sampled using a cytology brush for targeted metabolomic analysis by mass spectrometry. Multivariate analyses clustered heifers according to ovarian, uterine, and hormonal variables in clusters A (n = 5) and B (n = 8 heifers). Individual metabolite concentrations were compared between clusters A and B by univariate analysis using t-test after FDR correction. Concentrations of Pro, Ala, Leu, Gly, Val, Lys, Ile, Phe, Asp, Orn, Tyr, Arg, Trp, Suc, Cit, ADMA, the sum of essential Amino Acids (AA), sum of non-essential AA, sum of aromatic AA, and total AA were greater in cluster A (FDR ≤ 0.05). ULF volume dynamics and associated uterine, ovarian, and hormonal variables during the peri-estrus period presented a concerted variation among heifers, which was associated with the ULF composition four days after estrus. Potential implications for embryo receptivity and reproductive outcomes are the focus of the current investigation.
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