18 research outputs found
Dysregulated placental microRNAs in Early and Late onset Preeclampsia
Copyright © 2017. Published by Elsevier Ltd.INTRODUCTION: To determine the miRNA expression profile in placentas complicated by Preeclampsia (PE) and compare it to uncomplicated pregnancies. METHODS: Sixteen placentas from women with PE, [11 with early onset PE (EOPE) and 5 with late onset PE (LOPE)], as well as 8 placentas from uncomplicated pregnancies were analyzed using miRNA microarrays. For statistical analyses the MATLAB® simulation environment was applied. The over-expression of miR-518a-5p was verified using Quantitative Real-Time Polymerase Chain Reaction. RESULTS: Forty four miRNAs were found dysregulated in PE complicated placentas. Statistical analysis revealed that miR-431, miR-518a-5p and miR-124* were over-expressed in EOPE complicated placentas as compared to controls, whereas miR-544 and miR-3942 were down-regulated in EOPE. When comparing the miRNA expression profile in cases with PE and PE-growth restricted fetuses (FGR), miR-431 and miR-518a-5p were found over-expressed in pregnancies complicated by FGR. DISCUSSION: Since specific miRNAs can differentiate EOPE and LOPE from uncomplicated placentas, they may be considered as putative PE-specific biomarkers. MiR-518a-5p emerged as a potential diagnostic indicator for EOPE cases as well as for PE-FGR complicated placentas, indicating a potential link to the severity of the disease.Peer reviewe
Investigation of FANCA mutations in greek patients
Background: Fanconi anemia (FA) is a rare genetic disease characterized by considerable heterogeneity. Fifteen subtypes are currently recognised and deletions of the Fanconi anemia complementation group A (FANCA) gene account for more than 65% of FA cases. We report on the results from a cohort of 166 patients referred to the Department of Medical Genetics of Athens University for genetic investigation after the clinical suspicion of FA. Materials and Methods: For clastogen-induced chromosome damage, cultures were set up with the addition of mitomycin C (MMC) and diepoxybutane (DEB), respectively. Following a positive cytogenetic result, molecular analysis was performed to allow identification of causative mutations in the FANCA gene. Results: A total of 13/166 patients were diagnosed with FA and 8/13 belonged to the FA-A subtype. A novel point mutation was identified in exon 26 of FANCA gene. Conclusion: In our study 62% of FA patients were classified in the FA-A subtype and a point mutation in exon 26 was noted for the first time
Molecular diagnosis of Fragile X syndrome
Fragile X syndrome, the most prevalent inherited cause of mental
retardation, is related to hyperexpansion of a polymorphic CGG repeat of
the FMR1 gene. Expansion of 55-200 repeats are called premutations and
characterize carriers who usually have no mental impairment. The disease
causing full mutations exceed 200 CGG repeats, are hypermethylated and
lead to transcriptional silencing of the gene and absence of the Fragile
X mental retardation protein (FMRP). Diagnostic approaches involve
molecular and immunocytochemical techniques. Southern blot, which allows
mutations to be detected and methylation status to be determined in a
single test, remains the procedure of choice for most laboratories.
Modifications of PCR methods, including methylation specific PCR, are
also proposed but their implementation is still in question because of
inherent difficulties to amplify CGG repeats, distinguish between mosaic
patterns and interpret results in female individuals. The FMRP antibody
test is also suitable for large population screening and elucidation of
Fragile X syndrome cases with no CGG expansion, but it is not widely
applied. In search for novel diagnostic approaches, use of PCR as a
first prescreening test followed by Southern blot is considered the most
reliable procedure
Urine proteomic studies in preeclampsia
Preeclampsia (PE) is a multisystem disorder of pregnancy that develops
after 20 wk of gestation in previously normotensive women and
complicates 5-8% of pregnancies. This rapidly progressive syndrome is
usually diagnosed when the mother develops hypertension and proteinuria.
The only effective treatment is delivery of the baby although early
low-dose aspirin has been shown to significantly reduce the risk for PE.
Recent advances in proteomic methods of protein separation,
identification, and quantitation may allow for the identification of
proteins and peptides that could facilitate early detection of disease,
improve assessment of prognosis, and allow closer monitoring of women at
risk for PE. This review summarizes all currently available markers for
prediction and diagnosis of PE and presents urine proteomic studies
performed for the identification of novel biomarkers
Application of proteomics for diagnosis of fetal aneuploidies and pregnancy complications
Proteomic technologies represent new strategies towards high-throughput,
simultaneous analysis of thousands of biological molecules leading to
the discovery of biomarkers for early diagnosis, prognosis and
prediction of pregnancy outcome. Proteomics have additional relevance in
understanding pathophysiology and the development of molecularly
targeted therapeutics. Comparison of normal human amniotic fluid
proteome with that coming from pregnancies carrying fetuses with
chromosomal abnormalities facilitated the detection of panels of
potential biomarkers for prenatal detection of fetal aneuploidies.
Candidate biomarkers for the early prediction of preeclampsis are also
available, while four biomarkers (defensins-2 and -1, calgranulin-C, and
calgranulin-A), which were called the “MR score”, can quickly and
accurately detect potentially dangerous infections and predict premature
birth. Researchers remain hopeful that proteomic studies will allow for
the identification of either one protein marker or of a panel of markers
for prenatal detection of fetal aneuploidies and pregnancy complications
that could be usefully employed for diagnostic purposes or improvement
of the current screening methods. For maximum predictive power however,
biomarkers should be selected for further comparative analysis of
expression and structural modifications in large numbers of samples from
chromosomally normal and abnormal pregnancies obtained from different
populations. (C) 2009 Elsevier B.V. All rights reserved
First Trimester Maternal Plasma Aberrant miRNA Expression Associated with Spontaneous Preterm Birth
Spontaneous Preterm Delivery (sPTD) is one of the leading causes of perinatal mortality and morbidity worldwide. The present case–control study aims to detect miRNAs differentially expressed in the first trimester maternal plasma with the view to identify predictive biomarkers for sPTD, between 320/7 and 366/7 weeks, that will allow for timely interventions for this serious pregnancy complication. Small RNA sequencing (small RNA-seq) of five samples from women with a subsequent sPTD and their matched controls revealed significant down-regulation of miR-23b-5p and miR-125a-3p in sPTD cases compared to controls, whereas miR-4732-5p was significantly overexpressed. Results were confirmed by qRT-PCR in an independent cohort of 29 sPTD cases and 29 controls. Statistical analysis demonstrated that miR-125a is a promising early predictor for sPTL (AUC: 0.895; 95% CI: 0.814-0.972; p < 0.001), independent of the confounding factors tested, providing a useful basis for the development of a novel non-invasive predictive test to assist clinicians in estimating patient-specific risk
Validation of Serum Biomarkers Derived from Proteomic Analysis for the Early Screening of Preeclampsia
Aim. To examine the potential value of previously identified biomarkers using proteomics in early screening for preeclampsia (PE). Methods. 24 blood samples from women who subsequently developed PE and 48 from uncomplicated pregnancies were obtained at 11–13 weeks and analysed after delivery. Cystatin-C, sVCAM-1, and Pappalysin-1 were quantified by ELISA. Maternal characteristics and medical history were recorded. Results. Median values of Cystatin-C, sVCAM-1, and Pappalysin-1 in the PE group as compared to controls were 909.1 gEq/mL versus 480.0 gEq/mL, P=.000, 832.0 gEq/mL versus 738.8 gEq/mL, P=.024, and 234.4 gEq/mL versus 74.9 gEq/mL, P=.064, respectively. Areas under the receiver-operating characteristic curves (AUC, standard error (SE)) for predicting PE were Cystatin-C: 0.90 (SE 0.04), VCAM-1: 0.66 (SE 0.074), and Pappalysin-1: 0.63 (SE 0.083). To discriminate between cases at risk for PE and normal controls, cut-off values of 546.8 gEq/mL for Cystatin-C, 1059.5 gEq/mL for sVCAM-1, and 220.8 gEq/mL for Pappalysin-1 were chosen, providing sensitivity of 91%, 41%, and 54% and specificity of 85%, 100%, and 95%, respectively. Conclusions. sVCAM-1 and Pappalysin-1 do not improve early screening for PE. Cystatin-C, however, seems to be associated with subsequent PE development, but larger studies are necessary to validate these findings
Proteomic analysis of amniotic fluid in pregnancies with Down syndrome
Proteomic analysis is widely used for the detection of diagnostic markers. In the present study amniotic fluid supernatants?(AFS) from pregnancies with Down syndrome?(DS) fetuses and from chromosomally normal fetuses in the 17th?week of gestation were analyzed by 2-DE. Gel comparison revealed significant differences in the two groups. Spots with different expression levels were excised and proteins were identified by MALDI-MS and nano-ESI-MS/MS. Splicing factor arginine/serine-rich?4 (SFRS4; Q08170) was present only in AFS from DS fetuses and completely absent in the control group. Quantitative differences were detected for alpha-1-microglobulin (AMBP; P02760), collagen alpha?1?(I) chain (CO1A1; P02452), collagen alpha?1?(III) chain (CO3A1; P02461), collagen alpha?1?(V) chain?d (CO5A1; P20908), and basement membrane-specific heparin sulfate proteoglycan core protein (PGBM; P98160). These proteins were increased in cases with DS, whereas protein?IBP-1 (P08833) was decreased by 40% compared with chromosomally normal fetuses. Four proteins, CO1A1, CO3A1, CO5A1, and PGBM, appeared as fragments. As differentially expressed proteins were present in all pregnancies with DS tested, they may represent useful potential markers for prenatal diagnosis. However, for protein biomarkers to be of any clinical utility, systematic analysis of the maternal serum should be conducted