The Role of Circulating Biomarkers in the Early Diagnosis of Ovarian Cancer

Ovarian cancer is the leading cause of gynecologic-related cancer death and epithelial ovarian cancer (EOC) is the most lethal sub-type. EOC is usually asymptomatic, and few screening tests are available. Diagnosis of ovarian cancer can be difficult because of the nonspecific symptoms. Despite the various diagnostic methods used, there is no reliable early diagnostic test and it needs to be developed. Specific biomarkers may have potential with the least possible invasive procedure. Biomarkers with a high sensitivity to ovarian cancer should be identified. Circulating biomarkers that are significant tools for non-invasive early diagnosis can be analyzed using circulating tumor cells, exosomes, and circulating nucleic acids. Protein, gene, metabolite, and miRNA-based biomarkers can be used for ovarian cancer diagnosis. As non-coding RNAs, MiRNAs may have an important role in ovarian cancer diagnosis due to their effects on mRNA expression levels. The most recent developments regarding the potential of circulating biomarkers to detect early ovarian cancer is presented in this chapter

Potential function of microRNAs in thoracic aortic aneurysm and thoracic aortic dissection pathogenesis.

Torasik Aortik Anevrizma (TAA) ve Torasik Aortik Diseksiyon (TAD) 'sessiz katiller' olarak bilinen aort hastalıklarıdır. TAA, normal aort çapının en az yarısı kadar bir büyüme ile karakterize edilirken, TAD, aort duvar katmanlarının kademeli olarak ayrılmasıyla sonuçlanan yalancı lümen oluşumuyla karakterize edilir. Bu çalışmada, dokuz adet TAA'lı, dokuz adet TAD'lı hasta ile on adet sağlıklı bireyden oluşan toplam 28 serum örneği incelenmiştir. Bu çalışmanın amacı, TAA ve TAD patogenezinden sorumlu olan aday miRNA'ları tanımlamak için, hsa-miR-143-3p ve hsa-miR22-3p’ün ekspresyon profillerini araştırmaktır. Aday miRNA'larının hedefledikleri mRNA’ların biyoinformatik araçlar ile tespit edilmesinin ardından, hedef mRNA'ların ekspresyon profilleri analiz edildi. Kirsten sıçan sarkoması viral onkojen homologu (KRAS), mitojenle aktive edilmiş protein kinaz (MAPK) 7, MAPK14 ve transgelin (TAGLN) mRNA ekspresyon profillerini kantitatif polimeraz zincir reaksiyonu ile tespit edildi. Rölatif karşılaştırma sonucu, hsa-miR-143-3p (P = 0.017) ve hsa-miR-22-3p (P = 0.03) aday miRNA'larının ekspresyon sevilerindeki artış, TAA grubunda istatistiksel olarak anlamlı iken, TAD grubunda istatistiksel olarak anlamlı değildir. Kontrol grubu ile karşılaştırıldığında, KRAS ve MAPK7 mRNA'larının ekspresyon seviyesi her iki grupta da azalmıştır. Kontrol grubu ile karşılaştırıldığında MAPK14'ün ekspresyon seviyesi TAD grubunda azalmış, ancak TAA grubunda arttmıştır. TAGLN mRNA ekspresyon seviyesi iki grupta da artmıştır. Hsa-miR-143-3p‘nin anlatımındaki istatistiksel olarak anlamlı fark, hedef mRNA olan KRAS ve MAPK7‘nin anlatımlarının azalması ve miRNA-mRNA arasındaki negatif korelasyon, TAA için potansiyel biyobelirteç olabileceğini düşündürmektedir. Bu miRNA’lar ve ilişkili genleri TAA oluşumunda önemli fonksiyonlara hizmet edebilir, anlatımı değişmiş miRNA'lar TAA patogenezinde önemli olabilir

Detection of fetal RHD pseudogene (RHD Psi) and hybrid RHD-CE-D-s from RHD-negative pregnant women with a free DNA fetal kit

Hemolytic disease of the newborn is a clinical condition in which maternal and paternal Rh blood group antigens are incompatible and the mother is negative for the antigen whereas the father is positive. Analysis of fetal cells recovered from maternal plasma can provide a highly sensitive prenatal diagnosis. The fetal RHD gene in plasma DNA is detected by real-time PCR amplification of two different segments of the RHD gene (exons 7 and 10). Each amplicon is revealed with specific probes. We examined 40 female blood samples to verify the specificity of RHD exons (7 and 10) amplified by real-time PCR. Thirty fetuses were predicted to be RHD-positive based on analysis of plasma DNA. Seven fetuses were predicted to be RHD-negative. One fetus was negative for RHD on exon 10, and positive for RHD on exon 7 (early gestation age); two fetuses were RHD-negative on exon 7, and RHD- positive on exon 10 (RHD-CE-D-s or RHD Psi), indicative of a maternal RHD allele. We conclude that it is necessary to analyze at least two exon regions in the RHD gene

LARGE SCALE PRE-DIAGNOSIS OF TOXOPLASMA GONDII DNA GENOTYPING BY REAL-TIME PCR ON AMNIOTIC FLUID

The antepartum and peripartum maternal infections cause great problems complicating pregnancy. The early diagnosis of the maternal infections causing different pathologies in the newborns is of great importance. For an appropiate judgement, the early diagnosis of Toxoplasma gondii infections acquired during pregnancy is critical for an effective prevention. The common characteristics of these infections are that they may cause abortion, still birth, prematurity, intrauterin growth retardation, congenital malformations, infection of the newborn or normal term live-births. We report here the development of real-time PCR-based assay for detection of T gondii. Investigation of 300 specimens was carried out using B1 gene region specific primers and probes after the extraction of T gondii DNA. T.gondii DNA, was found in 4 (1.3%) out of 300 specimens. DNA was not found in the speciemens of the remaining 296 patients. Real-time PCR analysis significantly improves the detection of T gondii in amniotic fluid

MicroRNA expression profiling in placenta and maternal plasma in early pregnancy loss

Early pregnancy loss (EPL), also termed early miscarriage, is determined as the unintentional expulsion of an embryo or fetus prior to the 12th week of gestation. EPL frequency is similar to 15% in pregnancies. Fetal development and growth is associate with placental function and vessel development; therefore, the placental genome would represent a useful miscarriage model for (epi)genetic and genomic studies. An important factor of placental development and function is epigenetic regulation of gene expression. microRNAs (miRNAs) are the primary epigenetic regulators which have an important role in placental development and function. In the present study, maternal plasma and villous tissue were collected from 16 EPL cases in 6th-8th gestational weeks (GWs) and 8 abortions (control group) in 6th-8th GWs. Detection of the differences in miRNA expression was performed using microarrays and dysregulated miRNAs were validated by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). miRNA microarray findings revealed that four miRNAs, including hsa-miRNA (miR)-125a-3p, hsa-miR-3663-3p, hsa-miR-423-5p and hsa-miR-575 were upregulated in tissue samples. In maternal plasma, two miRNAs (hsa-let-7c, hsa-miR-122) were upregulated and one miRNA (hsa-miR-135a) was downregulated. A total of 6 out of 7 dysregulated miRNAs were validated using RT-qPCR. The target genes of these dysregulated miRNAs were detected using the GeneSpring database. The aim of the present study was to detect dysregulated miRNAs in maternal plasma and villous cells and identify the target genes of dysregulated miRNAs and their associated pathways. The target gene analyses have revealed that the affected genes are primarily associated with cell migration, proliferation, implantation, adhesion, angiogenesis and differentiation and all are involved with EPL pathogenesis. Therefore, the present study may contribute to the understanding of the molecular mechanisms which lead to EPL