UPLC-Q-TOF/MS based Untargeted Metabolite and Lipid Analysis on Premature Ovarian Insufficiency Plasma Samples.

Background: Metabolomics is one of the main areas to understand cellular process at molecular level by analyzing metabolites. In recent years metabolomics has been emerged as key tool to understand molecular basis of disease, find diagnostic and prognostic biomarkers, and develop new treatment opportunities and drug molecules. Objective: In this study, an untargeted metabolite and lipid analysis were performed to identify potential biomarkers on premature ovarian insufficiency plasma samples. 43 POI subject plasma samples were compared with 32 healthy subject plasma samples. Methods: Plasma samples were pooled and extracted using chloroform:methanol:water (3:3:1 v/v/v) mixture. Agilent 6530 LC/MS Q-TOF instrument equipped with ESI source was used for analysis. A C18 column (Agilent Zorbax 1.8 μM, 50 x 2.1 mm) was used for separation of metabolites and lipids. XCMS, an “R software” based freeware program, was used for peak picking, grouping and comparing the findings. Isotopologue Parameter Optimization (IPO) software was used in order to optimize XCMS parameters. The analytical methodology and data mining process were validated according to the literature. Results: 83 metabolite peaks and 213 lipid peaks were found to be in semi-quantitatively and statistically different (fold change >1.5, p <0.05) between the POI plasma samples and control subjects. Conclusion: According to the results, two groups were successfully separated through principal component analysis. Among the peaks, phenyl alanine, decanoyl-L-carnitine, 1-palmitoyllysophosphatidylcholine and PC(O-16:0/2:0) were identified through auto MS/MS and matched with human metabolome database and proposed as plasma biomarker for POI and monitoring the patients in treatment period.

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Analysis of in vitro follicle development during the onset of premature ovarian insufficiency in a mouse model

Reproduction Fertility and Development ◽

10.1071/rd15524 ◽

2017 ◽

Vol 29 (8) ◽

pp. 1538 ◽

Cited By ~ 1

Author(s):

Heidy Kaune ◽

Sairah Sheikh ◽

Suzannah A. Williams

Keyword(s):

Mouse Model ◽

Premature Ovarian Insufficiency ◽

Controlled Environment ◽

Follicle Development ◽

Growth Morphology ◽

Follicle Growth ◽

Ovarian Insufficiency ◽

Female Mice ◽

And Control

Premature ovarian insufficiency (POI) occurs in 1% of women under 40 years of age and is predominantly idiopathic. In a transgenic mouse model of follicular POI, the Double Mutant (DM), female mice are fertile at 6 weeks of age, become infertile by 9 weeks and exhibit POI by 3 months. DM female mice generate oocytes lacking mucin O-glycans and complex N-glycans due to deletion of core 1 synthase, glycoprotein-N-acetylgalactosamine 3-β-galactosyltransferase 1 (C1galt1) and mannoside acetylglucosaminyltransferase 1 (Mgat1) respectively (DM, C1galt1F/FMgat1F/F:ZP3Cre; Control, C1galt1F/FMgat1F/F). To determine whether DM follicle development could be improved in a controlled environment, follicles from DM and Control mice were cultured individually and follicle growth, morphology, survival and antrum formation were evaluated. DM ovaries were more rigid than Control ovaries at 3, 6 and 9 weeks, which was exacerbated with age, resulting in a failure to isolate follicles from 9 week-old DM females. DM follicles had decreased survival compared with Control follicles from females at 3 and 6 weeks of age. Furthermore, survival rate of DM follicles decreased with age between 3 and 6 weeks. DM follicles at both 3 and 6 weeks had accelerated follicle growth and altered antrum formation during the first few days of culture but, after 6 days, follicles were equivalent in size to the Controls. In conclusion, a population of DM follicles retain the potential to develop in vitro, and therefore follicle culture offers a reliable method to generate antral follicles from preantral follicles after the onset of POI in these female mice.

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Metabolomics-Based Study of Clinical and Animal Plasma Samples in Coronary Heart Disease with Blood Stasis Syndrome

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2012/638723 ◽

2012 ◽

Vol 2012 ◽

pp. 1-6 ◽

Cited By ~ 6

Author(s):

Huihui Zhao ◽

Jianxin Chen ◽

Qi Shi ◽

Xueling Ma ◽

Yi Yang ◽

...

Keyword(s):

Coronary Heart Disease ◽

Heart Disease ◽

Blood Stasis ◽

Principal Component ◽

Blood Stasis Syndrome ◽

Control Group ◽

Swine Model ◽

Plasma Samples ◽

Healthy Control ◽

And Control

The aim of this study is to explore a bridge connecting the mechanism basis and macro syndromes of coronary heart disease with experimental animal models. GC-MS technique was used to detect the metabolites of plasma samples in mini swine models with myocardial infarction (MI) and patients with unstable angina (UA). 30 metabolites were detected in the plasma samples of more than 50 percent of model group and control group in swine, while 37 metabolites were found in the plasma samples of UA patients and healthy control group. 21 metabolites in the plasma samples of swine model and 20 metabolites in patients with UA were found of significant value. Among which, 8 shared metabolites were found of low level expression in both swine model and UA patients. Independent Student’st-test, principal component analysis (PCA), and hierarchicalcluster analysis (HCA) were orderly applied to comprehend inner rules of variables in the data. The 8 shared metabolites could take place of the 21 or 20 metabolites in classification of swine model with MI and UA patients, which could be considered as a bridge connecting the mechanism basis and macrosyndromes of swine model with MI and UA patients.

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How polymorphic markers contribute to genetic diseases in different populations? The study of inhibin A for premature ovarian insufficiency

Einstein (São Paulo) ◽

10.1590/s1679-45082017ao4052 ◽

2017 ◽

Vol 15 (3) ◽

pp. 269-272 ◽

Cited By ~ 2

Author(s):

Denise Maria Christofolini ◽

Emerson Barchi Cordts ◽

Fernando Santos-Pinheiro ◽

Erika Azuma Kayaki ◽

Mayla Cristina Fernandes Dornas ◽

...

Keyword(s):

Genetic Diseases ◽

Premature Ovarian Insufficiency ◽

Polymorphic Markers ◽

Ovarian Insufficiency ◽

Exon 2 ◽

Pcr Rflp ◽

Different Populations ◽

Study Participants ◽

And Control ◽

Edta Tube

ABSTRACT Objective To verify the incidence of the G679A mutation in exon 2 of the gene inhibin alpha (INHA), in women with secondary amenorrhea and diagnosis of premature ovarian insufficiency, and in controls. Methods A 5mL sample of peripheral blood was collected from all study participants in an EDTA tube and was used for DNA extraction. For the patient group, 5mL of blood were also collected in a tube containing heparin for karyotype, and 5mL were collected in a dry tube for follicle stimulant hormone dosage. All patient and control samples were initially submitted to analysis of the G679A variant in exon 2 of the INHA gene by PCR-RFLP technique. Samples from patients with premature ovarian insufficiency after PCR-RFLP were submitted to Sanger sequencing of the encoding exons 2 and 3. Sequencing was performed on ABI 3500 GeneticAnalyzer equipment and the results were evaluated by SeqA and Variant Reporter software. Results Samples of 70 women with premature ovarian insufficiency and 97 fertile controls were evaluated. The G769A variant was found in only one patient in the Premature Ovarian Insufficiency Group and in no control, and it appears to be rare in Brazilian patients with premature ovarian insufficiency. This polymorphism was previously associated to premature ovarian insufficiency in several populations worldwide. Conclusion There is genetic heterogeneity regarding the INHA gene in different populations, and among the causes of premature ovarian insufficiency.

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