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They revealed that potential biomarkers involved in idiopathic preterm labor and delivery included five SELDI peaks in amniotic fluid (AF) Q-profile, placenta villous hypermaturation and senescence-associated β-galactosidase activity in the placenta, and protein Z levels and FokI Vitamin D receptor polymorphism in maternal blood 6, 7, 8, 9. Few studies have been carried out to explore single or a few biomarkers as potential etiologic agents of idiopathic preterm labor and delivery. Despite the clinical relevance of idiopathic PTL for considering subfractions of prevalence, this area of study has received little attention from investigators involved in biomarker research mainly because the definition of idiopathic PTL subgroups using objective criteria has not yet been determined. While approximately 40% of SPTD cases are associated with mostly subclinical inflammations/infections, more than half of the SPTD cases are of unknown etiology (idiopathic, meaning that no identifiable cause is found) 5. Therefore, identifying biomarkers for biochemical changes that may be causally linked to SPTD in cases other than infection/inflammation is important for implementing mechanism-based treatment and prevention of PTL followed by SPTD. To date, the only PTL mechanism that has been causally linked to spontaneous preterm delivery (SPTD) is infection/inflammation 4, whereas the others are mostly implicated by associations suggested by epidemiologic, clinical, experimental, or placental pathologic observations 3.

PTL is regarded as a complex syndrome with multiple etiologies that include genetic predisposition, infection and/or inflammation, decidual hemorrhage and vascular disease, hormonal dysregulation, and uterine overdistension 3. Preterm labor with intact membrane (PTL) complicates 2–3% of pregnancies and accounts for approximately one-third of preterm births, causing significant neonatal mortality and morbidity 1, 2. The proteomic analysis data of AFs from women with PTL identified several novel biomarkers and specific protein pathways related to SPTD in the absence of IUI. Five principle pathways associated with the 77 DEPs were identified, including glycolysis, gluconeogenesis, and iron homeostasis. ELISA validation confirmed that women who had an SPTD before 34 weeks had significantly independently lower levels of VEGFR-1 and higher levels of lipocalin-2 and the Fc fragment of IgG binding protein in the AF. Proteomic analysis identified 77 differentially expressed proteins (DEPs, P < 0.05) in the AF from SPTD cases compared to term delivery controls. In the total cohort, an ELISA validation study was performed for seven candidate proteins of interest. A nested case–control was conducted using pooled AF samples (n = 20) analyzed via label-free liquid chromatography-tandem mass spectrometry. This was a retrospective cohort study of a total of 139 PTL women with singleton gestation (24 + 0 to 32 + 6 weeks) who underwent amniocentesis and who displayed no evidence of IUI. We sought to identify biomarkers in the amniotic fluid (AF) and specific signaling pathways related to spontaneous preterm delivery (SPTD, < 34 weeks) in women with preterm labor (PTL) without intra-uterine infection/inflammation (IUI).