RESUMEN
Gestational Diabetes Mellitus (GDM) causes severe short- and long-term complications for the mother, fetus and neonate, including type 2-diabetes (T2DM) later in life. In this pilot study, GC-Q/MS analysis was applied for plasma metabolomics fingerprinting of 24 healthy and 24 women with GDM at different stages of gestation (second and third trimester) and postpartum (one and three months). Multivariate (unsupervised and supervised) statistical analysis was performed to investigate variance in the data, identify outliers and for unbiased assessment of data quality. Plasma fingerprints allowed for the discrimination of GDM pregnant women from controls both in the 2nd and 3rd trimesters of gestation. However, metabolic profiles tended to be similar after delivery. Follow up of these women revealed that 4 of them developed T2DM within 2 years postpartum. Multivariate PLS-DA models limited to women with GDM showed clear separation 3 months postpartum. In the 2nd trimester of gestation there was also a clear separation between GDM women that were normoglycemic after pregnancy and those with recognized postpartum T2DM. Metabolites that had the strongest discriminative power between these groups in the 2nd trimester of gestation were 2-hydroxybutyrate, 3-hydroxybutyrate, and stearic acid. We have described, that early GDM comprises metabotypes that are associated with the risk of future complications, including postpartum T2DM. In this pilot study, we provide evidence that 2-hydroxybutyrate and 3-hydroxybutyrate may be considered as future prognostic biomarkers to predict the onset of diabetic complications in women with gestational diabetes after delivery.
Asunto(s)
Diabetes Gestacional , Ácido 3-Hidroxibutírico , Biomarcadores , Femenino , Cromatografía de Gases y Espectrometría de Masas , Humanos , Estudios Longitudinales , Proyectos Piloto , Embarazo , PronósticoRESUMEN
Gestational Diabetes Mellitus (GDM) is defined as glucose intolerance with onset or first recognition during pregnancy. It is affecting approximately up to 14% of all pregnancies with an increasing tendency. GDM has been related to relevant short-term and long-term health complications for both mother and offspring. Recent studies strongly emphasized the role of several essential amino acids in the pathogenesis of obesity and highlighted their strong correlation with insulin resistance, but there are no references related to modifications in D-AAs in biological fluids. As D-AA elimination proceeds mainly by renal excretion, urine was the selected sample to evaluate the alterations in free D-AAs ratio in a GDM study. Only 1 mL of first void urine or standard solution was required for purification, by using a Discovery DSC-SCX SPE cartridge (500 mg/3 mL) and derivatization into their N(O)-pentafluoropropionyl amino acid 2-propyl esters. Enantiomeric separation was carried out by GC-MS on a Chirasil-L-Val N-propionyl-L-valine-tert-butylamide polysiloxane fused-silica capillary column (25 m×0.25 mm I.D., 0.12 µm film thickness, Agilent Technologies, Waldbronn, Germany), under programmed temperature elution. Detection was performed with an ion trap mass analyzer, operating in the full scan mode in the m/z 50-350 range. 14 pairs of derivatives of D-and L-AAs were separated. The steps of sample preparation, derivatization and GC-MS conditions were optimized for both urine and standards. Several conditions affecting the SPE procedure, such as sorbent mass/volume ratio of the cartridge, sample dilution and pH, were optimized. Volume of reagents and solvents and reaction temperature and time were also tested for the derivatization. Regarding the GC-MS parameters, split ratio, temperature program and mass range were optimized. The final method was validated in terms of linearity, sensitivity, accuracy and precision for D-Ala, D-Pro, D-Ser, D-Met, D-Phe, D-Glu, D-Orn and D-Lys. Identification of AAs in urine samples was based on retention time and mass spectra. Urine from 20 women with GDM and 20 pregnant women with normal glucose tolerance (after 2-h 75-g oral glucose tolerance test), matched according to the week of gestation and age (22-28 week of gestation and age 24-37 years), were enrolled into the study. %D-Relative amounts were determined for Ala, Val, Thr, Ser, Leu, Asx (Asp+Asn), Glx (Glu+Gln), Met, Phe, Tyr, Orn and Lys. Statistically significant differences (p<0.05) were observed only for D-Phe and higher values were found in the GDM group. It is possible that D-Phe could be involved in metabolic/signaling pathways to compensate early stages of insulin resistance, although further work is necessary to confirm this hypothesis.
Asunto(s)
Aminoácidos/química , Aminoácidos/orina , Diabetes Gestacional/orina , Adulto , Femenino , Cromatografía de Gases y Espectrometría de Masas/métodos , Prueba de Tolerancia a la Glucosa/métodos , Humanos , Redes y Vías Metabólicas/fisiología , Embarazo , Transducción de Señal/fisiología , Adulto JovenRESUMEN
Gestational Diabetes (GDM) is causing severe short- and long-term complications for mother, fetus or neonate. As yet, the metabolic alterations that are specific for the development of GDM have not been fully determined, which also precludes the early diagnosis and prognosis of this pathology. In this pilot study, we determine the metabolic fingerprint, using a multiplatform LC-QTOF/MS, GC-Q/MS and CE-TOF/MS system, of plasma and urine samples of 20 women with GDM and 20 with normal glucose tolerance in the second trimester of pregnancy. Plasma fingerprints allowed for the discrimination of GDM pregnant women from controls. In particular, lysoglycerophospholipids showed a close association with the glycemic state of the women. In addition, we identified some metabolites with a strong discriminative power, such as LPE(20:1), (20:2), (22:4); LPC(18:2), (20:4), (20:5); LPI(18:2), (20:4); LPS(20:0) and LPA(18:2), as well as taurine-bile acids and long-chain polyunsaturated fatty acid derivatives. Finally, we provide evidence for the implication of these compounds in metabolic routes, indicative of low-grade inflammation and altered redox-balance, that may be related with the specific pathophysiological context of the genesis of GDM. This highlights their potential use as prognostic markers for the identification of women at risk to develop severe glucose intolerance during pregnancy. BIOLOGICAL SIGNIFICANCE: Gestational Diabetes Mellitus (GDM) is increasing worldwide and, although diabetes usually remits after pregnancy, women with GDM have a high risk of developing postpartum type 2-diabetes, particularly when accompanied by obesity. Therefore, understanding the pathophysiology of GDM, as well as the identification of potentially modifiable risk factors and early diagnostic markers for GDM are relevant issues. In the present study, we devised a multiplatform metabolic fingerprinting approach to obtain a comprehensive picture of the early metabolic alternations that occur in GDM, and may reflect on the specific pathophysiological context of the disease. Future studies at later stages of gestation will allow us to validate the discriminant power of the identified metabolites.
Asunto(s)
Diabetes Gestacional/fisiopatología , Adulto , Biomarcadores/metabolismo , Glucemia/metabolismo , Diabetes Gestacional/sangre , Diabetes Gestacional/orina , Femenino , Intolerancia a la Glucosa/etiología , Humanos , Fosfolípidos/sangre , Proyectos Piloto , Embarazo , Segundo Trimestre del Embarazo , Pronóstico , ProteómicaRESUMEN
OBJECTIVE: The aim of this study was to investigate the cerebral circulation in pregnancy complicated with oligohydramnios. MATERIALS AND METHODS: Comparative clinical studies evaluated Doppler curve middle cerebral artery separately in proximal (basal, MCA-P) segment and distal (cortical, MCA-D) anatomical segment. Doppler velocity waveforms analysis was performed on the base of systolic (Vs), mean (Vm) and end-diastolic (Ved) velocity of the blood wave, systolic/diastolic ratio (S/D), resistance index (RI) and pulsatile index (PI). Results were compared in the group with oligohydramnios (15 pregnancies) and the group with normohydramnios (15 pregnancies). RESULTS: We found statistically significant correlation between S/D MCA-P, S/D MCA-D, RI MCA-P. Other parameters did not show statistically significant differences. Especially we did not find statistically significant differences of parameters of the fetal cerebral circulations between pregnancies with oligohydramnios and with normohydramnios. CONCLUSIONS: On the basis of this correlation, the following conclusions were drawn: 1/ pregnancies complicated with oligohydramnios do not change fetal cerebral circulation, 2/ idiopathic oligohydramnios does not qualify for group of high risk pregnancy.
Asunto(s)
Circulación Cerebrovascular , Arteria Cerebral Media/diagnóstico por imagen , Oligohidramnios/diagnóstico por imagen , Ultrasonografía Doppler Transcraneal , Ultrasonografía Prenatal , Velocidad del Flujo Sanguíneo , Estudios de Casos y Controles , Femenino , Retardo del Crecimiento Fetal/diagnóstico por imagen , Monitoreo Fetal , Humanos , Embarazo , Tercer Trimestre del Embarazo , Flujo Pulsátil , Sensibilidad y EspecificidadRESUMEN
OBJECTIVE: To compare the effect of misoprostol (PGE(1)) versus dinoprostone (PGE(2)) on blood flow in uteroplacental circulation during labor induction. STUDY DESIGN: Eighty-four women with indications for induction of labor were assigned to receive either misoprostol 50 microg per vagina every 4 h as needed or 0.5 mg doses of dinoprostone given intra-cervically every 6 h by means of a randomization table generated by computer. Doppler velocimetry of umbilical, uterine and arcuate arteries was performed immediately before and 2-3 h after the administration of misoprostol or dinoprostone. The SAS system was used to perform statistical analysis. RESULTS: There were no significant changes of pulsatility index (PI), resistance index (RI) and systolic/diastolic (S/D) ratio in umbilical arteries after both prostaglandin compounds. Vaginal application of misoprostol significantly increased all ratios in arcuate artery and S/D ratio in uterine artery. Intra-cervically dinoprostone significantly increased PI, RI and S/D ratio in arcuate and uterine arteries. CONCLUSIONS: Our results indicate that vaginal misoprostol and cervical dinoprostone administration increases uteroplacental resistance but does not affect umbilical blood flow. Misoprostol would be as safe and effective agent as dinoprostone for cervical ripening and labor induction.