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In humans and most animals, maternal inheritance of mitochondria and mitochondrial DNA (mtDNA) is considered as an universal assumption. Recently, several lines of evidence suggest that different species seem to employ distinct mechanisms to prevent the inheritance of paternal mtDNA. There are few studies in the literature on the molecular basis of sperm mtDNA elimination in mammals and paternal mtDNA transmission in humans. Endonuclease G (ENDOG) is a mitochondrial nuclease encoded by nuclear ENDOG gene. The critical importance of ENDOG gene on paternal mitochondrial elimination (PME) has been previously demonstrated in model organisms such as C. elegans and D. melanogaster. However, its mechanism in human is still unclear. Therefore, we aimed to evaluate whether nuclear ENDOG gene copy number could be a potential marker of paternal mtDNA transmission or not.Male factor infertility patients diagnosed with different infertility subgroups such as azoospermia, oligoteratozoospermia, astheno-teratozoospermia were included in this study: 13 infertile men and 25 healthy men as control group. Quantitative real-time polymerase chain reaction (qPCR) analysis and dual-color Fluorescence in situ hybridization (FISH) method were used to compare the groups. FISH method was applied to verify qPCR results and two signals were observed in nearly all patients. ENDOG gene copy number data were evaluated by comparing them with entire human mtDNA next-generation sequencing (NGS) analysis results obtained through bioinformatics and proteomics tools. Mitochondrial whole genome sequencing (WGS) data allowed determination of novel and reported variations such as single nucleotide polymorphisms (SNPs), multiple nucleotide polymorphism (MNP), insertion/deletion (INDEL). Missense variants causing amino acid substitution were filtered out from patients' mtDNA WGS data.Relative copy number of target ENDOG gene in male infertility patients [0.49 (0.31 - 0.77)] was lower than healthy controls [1.00 (0.66 - 1.51)], and statistical results showed significant differences between the groups (p 0.01). A total of 38 missense variants were detected in the genes encoding the proteins involved in the respiratory chain complex. Moreover, we detected paternal mtDNA transmissions in the children of these patients who applied to assisted reproductive techniques.In conclusion, this study reveals that ENDOG gene may be an important factor for the PME mechanism in humans. To the best of our knowledge, this is the first study in humans about this topic and assessment of ENDOG gene sequencing and gene expression studies in a larger sample size including patients with male factor infertility would be our future project.

Epithelial ovarian cancer (EOC) is the type of OC with the highest mortality rate. Due to the asymptomatic nature of the disease and few available diagnostic tests, it is mostly diagnosed at the advanced stage. Therefore, the present study aimed to discover predictive and/or early diagnostic novel circulating microRNAs (miRNAs or miRs) for EOC. Firstly, microarray analysis of miRNA expression levels was performed on 32 samples of female individuals: Eight plasma samples from patients with pathologically confirmed EOC (mean age, 45 (30-54) years), eight plasma samples from matched healthy individuals (HIs) (mean age, 44 (30-65) years), eight EOC tissue samples (mean age, 45 (30-54) years) and eight benign ovarian (mean age, 35 (17-70) years) neoplastic tissue samples A total of 31 significantly dysregulated miRNAs in serum and three miRNAs in tissue were identified by microarray. The results were validated using reverse transcription-quantitative PCR on samples from 10 patients with pathologically confirmed EOC (mean age, 47(30-54) years), 10 matched His (mean age, 40(26-65) years], 10 EOC tissue samples (mean age, 47(30-54) years) and 10 benign ovarian neoplastic tissue samples (mean age, 40(17-70) years). The 'Kyoto Encyclopedia of Genes and Genomes' (KEGG) database was used for target gene and pathway analysis. A total of three miRNAs from EOC serum (hsa-miR-1909-5p, hsa-miR-885-5p and hsa-let-7d-3p) and one microRNA from tissue samples (hsa-miR-200c-3p) were validated as significant to distinguish patients with EOC from HIs. KEGG pathway enrichment analysis showed seven significant pathways, which included 'prion diseases', 'proteoglycans in cancer', 'oxytocin signaling pathway', 'hippo signaling pathway', 'adrenergic signaling in cardiomyocytes', 'oocyte meiosis' and 'thyroid hormone signaling pathway', in which the validated miRNAs served a role. This supports the hypothesis that four validated miRNAs, have the potential to be a biomarker of EOC diagnosis and target for treatment.

It has been identified that endometrium specific microRNAs have different expression levels in endometrial tissues and maternal serum during endometrial cycle. The aim of this study was to analyze microRNA expression levels in recurrent implantation failure patients and healthy controls endometrial samples for enlightening the aetiopathogenesis of the disease. The second aim was to search for a potential noninvasive molecular biomarker in early diagnosis and treatment of Recurrent Implantation Failure (RIF) patients.Endometrium and serum samples in two different phases (PP; proliferative phase and SP; secretory phase) from the same cases (RIF;This study showed that has-miR-145, has-miR-23b, has-miR-31 and has-miR-30b were significantly up-regulated in PP and down-regulated in SP endometrium samples. In serum samples, has-miR-145 and hsa-miR-23b were significantly down-regulated in both of PP and SP. Target gene and pathway analysis for dysregulated miRNAs identified important, validated and predicted genes for the implantation process.This study is the first study to obtain endometrium and serum samples in two different phases from the same cases and measure the candidate miRNAs expression. Our finding suggests that expression level of four candidate miRNAs may be involved in RIF development in women. Furthermore, these miRNAs can be potential biomarker for early diagnosis of RIF patients.

Recent studies have focused on the early detection of ovarian cancer (OC) using tumor materials by liquid biopsy. The mechanisms of microRNAs (miRNAs) to impact OC and signaling pathways are still unknown. This study aims to reliably perform functional analysis of previously validated circulating miRNAs' target genes by using pathfindR. Also, overall survival and pathological stage analyses were evaluated with miRNAs' target genes which are common in the The Cancer Genome Atlas and GTEx datasets. Our previous studies have validated three downregulated miRNAs (hsa-miR-885-5p, hsa-miR-1909-5p, and hsa-let7d-3p) having a diagnostic value in OC patients' sera, with high-throughput techniques. The predicted target genes of these miRNAs were retrieved from the miRDB database (v6.0). Active-subnetwork-oriented Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis was conducted by pathfindR using the target genes. Enrichment of KEGG pathways assessed by the analysis of pathfindR indicated that 24 pathways were related to the target genes. Ubiquitin-mediated proteolysis, spliceosome and Notch signaling pathway were the top three pathways with the lowest p-values (p < 0.001). Ninety-three common genes were found to be differentially expressed (p < 0.05) in the datasets. No significant genes were found to be significant in the analysis of overall survival analyses, but 24 genes were found to be significant with pathological stages analysis (p < 0.05). The findings of our study provide in-silico evidence that validated circulating miRNAs' target genes and enriched pathways are related to OC and have potential roles in theranostics applications. Further experimental investigations are required to validate our results which will ultimately provide a new perspective for translational applications in OC management.

Introduction In preeclampsia (PE), human decidua mesenchymal stromal cells (hDMSCs) are exposed to abnormally high levels of oxidative stress and inflammatory factors circulating in the maternal blood. MicroRNAs (miRNAs) have been shown to have a significant impact on the differentiation, maturation and function of mesenchymal stromal cells (MSCs). Our aim in the present study is firstly to investigate differentially expressed miRNA levels to be used as a biomarker in the early detection of PE and secondly to investigate whether those differentially expressed miRNAs in hDMSCs have an effect on the pathogenesis of PE. Methods This study covers miRNA expression analysis of hDMSCs from 7 PE patient and 7 healthy pregnant women and is a preliminary study to investigate putative biomarkers. After cell culture and cell sorting, total RNA including miRNAs were isolated from hDMSCs. Let-7b-3p, let-7f-1-3p, miR-191–3p, miR-550a-5p, miR-33b-3p and miR-425–3p were used for miRNA analysis and U6 snRNA was used for normalization of the samples. MiRNA analysis was performed by droplet digital polymerase chain reaction (ddPCR) method and obtained results were evaluated statistically. Results As a result of the analysis, it was observed that the levels of hsa-miR-33b-3p significantly (AUC: 0.93, p = 0.04, fold change: 4.5) increased in hDMSC of PE patients compared to healthy controls. However, let-7b-3p, let-7f-1-3p, miR-191–3p, miR-550a-5p, and miR-425–3p were not considered as significant because they did not meet the p Discussion Within the scope of the study, it is predicted that miR-33b-3p (p = 0.004, AUC = 0.93) can be used as a biomarker in detecting PE.

The aim of this prospective cohort study was to identify altered biologic processes in the endometrium that may be potential markers of receptive endometrium in patients with repeated implantation failure (RIF) as compared with fertile controls. The study was conducted in a university-affiliated in vitro fertilization (IVF) gynecology clinic and molecular biology and genetics laboratory. Healthy fertile controls (n = 24) and patients with RIF (n = 24) were recruited. Window of implantation gene profiling associated with RIF was performed. Six hundred forty-one differentially expressed genes were identified, and 44 pathways were found enriched. Upon clustering of the enriched pathways, 9 representative pathways were established. The important pathways that were identified included circadian rhythm, pathways in cancer, proteasome, complement and coagulation cascades, citrate cycle, adherens junction, immune system and inflammation, cell cycle, and renin-angiotensin system. The involvement of the circadian rhythm pathway and other related pathways may alter the endometrium's functioning to ultimately cause RIF. Furthermore, we found that the pathogenesis of RIF was multifaceted and that numerous processes were involved. We believe that a better understanding of the underlying mechanisms of RIF will ultimately give rise to better treatment opportunities and to better outcomes in IVF.

BackgroundPreeclampsia (PE) is a multi-systemic disease of pregnancy and leading cause of maternal and fetal mortality and morbidity worldwide. The differential expression of circulating microRNAs (miRs) were reported in maternal plasma of pregnant women and they could be stabilized in plasma. The aim of this study was to characterize the molecular mechanism of PE development through miR-1183.Methods and ResultsPlasma samples were obtained between 26 and 32 weeks of gestation from early-onset PE cases (n=31) and women with normotensive pregnancies (controls, n=22). The expressions of miR-1183 and Churchill containing domain 1 (CHURC1) were measured in maternal plasma. CHURC1 3’ untranslated region was determined as the target of miR-1183 using in silico analysis. miR-1183 and CHURC1 expression levels were assessed by real-time polymerase chain reaction (RT-PCR). An inverse correlation was found between circulating miR-1183 and CHURC1 expression levels with, miR-1183 up-regulated (p=0.002) and CHURC1 down-regulated (p) in maternal plasma of preeclamptic women compared to controls. The accuracy of levels of miR-1183 and CHURC1 according to area under receiver operating curve (ROC) analysis were characteristic curve analysis: the area under the curve (AUC) were: 0.79; (CI: 0.62-0.91) and 0.96; (CI: 0.78-0.99), respectively.ConclusionThese findings not only characterize a new mechanism for the disease, but also provide potential therapeutic targets.

Objective: Benign ovarian cysts (BOC) are the most common tumors in women of reproductive age. Usually, these cysts are harmless, but, a small number of them occasionally progress to malignancy. Among ovarian malignancies, epithelial ovarian cancer (EOC) comprises 90\% and is the most important cause of gynecologic cancerrelated deaths. We aimed to identify dysregulated miRNAs in patients with benign ovarian cysts (n = 11) compared to EOC (n = 10) and to healthy individuals (HI) (n = 15). Methods: The serum samples from EOC and BOC patients were collected before operation. We studied three different sample groups (serum of EOC (n = 8), HI (n = 8), and BOC (n = 8) patients) that contained the highest-quality of RNA. Microarray data were analyzed according to expression of miRNAs and target genes by bioinformatics tools. Results: When compared to EOC samples, 75 miRNAs were dysregulated in BOC samples. Sixty-six miRNAs from BOC were dysregulated when compared to HI samples. Bioinformatics analysis of BOC vs. EOC and BOC vs. HI showed that 46 miRNAs were congruent and their expression alterations were similar (up- or down-regulated). Further analysis showed that these 46 miRNAs are associated to one of three pathways involved in cancer pathogenesis. Conclusion: Several miRNAs might playa role in BOC formation and/or malignant transformation. These dysregulated miRNAs could potentially be a biomarker to distinguish between a completely BOC and one that is malignant or has potential for malignant transformation.

Recurrent implantation failure (RIF) is diagnosed when good-quality embryos repeatedly fail to implant after transfer in several in vitro fertilization (IVF) treatment cycles. Different expression profiles in maternal mRNAs could be referring to many diseases including RIF. This study aimed to reveal significantly dysregulated selected genes expression between healthy fertile women and RIF patients in the implantation window days of the natural menstrual cycle. MME, WWC1, TNC, and FOXP3 genes were chosen as target genes regarding their possible relations with the implantation process. Pathways with these genes were identified and the relationship between these pathways and RIF was investigated. In this study, the endometrial biopsy samples were collected in the secretory phase (cycle day 20–24) of the menstrual cycle from RIF patients (n = 34) and healthy fertile controls (n = 34). After “Pathway and network-oriented GWAS analysis” (PANOGA) and “Kyoto Encyclopedia of Genes and Genomes” (KEGG) pathway analysis; “Membrane Metalloendopeptidase” (MME), “WW and C2 Domain Containing 1” (WWC1), “Tenascin C” (TNC) and “Forkhead Box P3” (FOXP3) genes were chosen as target genes by regarding their possible relation with implantation process. Detection of differences in mRNA expressions between the control group and RIF patients has been performed with the droplet digital PCR (ddPCR) method. Results of the study showed that MME and WWC1 genes expression levels are significantly (p

The people who make regular and professional sports have different structured and worked of hearts which is called the athlete’s heart...

Objective Aim of this study was to define the relationship between RIF (Recurrent Implantation Failure) and endometrial mtDNA copy number. Study Design A total of 50 women of reproductive age including twenty-five patients clinically diagnosed with RIF and twenty-five fertile women as healthy controls were recruited into the study. Endometrial biopsy samples were obtained with a pipelle at the 20–24 days of the menstrual cycle of each participant. Total genomic DNA samples were isolated from endometrial tissues; MT-ND1 (mitochondrially encoded NADH dehydrogenase I) and MT-CO2 (mitochondrially encoded cytochrome C oxidase II) target genes were amplified by droplet digital PCR (ddPCR). Nuclear GAPDH (Glyceraldehyde-3-Phosphate Dehydrogenase) gene was also used for study normalization. The study has been conducted between February 2019 and June 2019. Result(s) Droplet digital PCR results were analyzed in “QuantaSoft” software. The concentration amount (copies/μl) of each participant’s mitochondrial gene was normalized according to the GAPDH gene concentrations as nuclear reference. mtDNA amounts were compared between RIF patients and healthy controls. Normalized data was statistically evaluated using Mann-Whitney U test and ROC curve analysis. Conclusion(s) It was concluded that the mitochondrial target gene (MT-ND1 and MT-CO2) copy number amount of RIF patients was higher than the one obtained from the healthy group in endometrial tissues. It is thought that higher mtDNA copy number at the RIF group may be related to increased oxidative stress in the endometrium. This stress factors may influence receptivity negatively and cause implantation failure. The receptivity of the endometrium is associated with the number of mtDNA copies and difference can be used as a biomarker for receptivity analysis.

Purpose: Preeclampsia (PE) is a pregnancy specific disease soon after 20 weeks of gestation where major symptoms are hypertension and proteinuria. The underlying pathology is believed to be...

Preeclampsia (PE) is one of the leading causes of maternal and fetal morbidity and mortality, occurring usually in the second half of pregnancy and affecting approximately 5–8% of pregnancies in the world. miRNAs play critical role in the regulation of placental development processes. We aimed to determine specific novel miRNAs for early diagnosis of preeclampsia which is one of the most dangerous pregnancy diseases. In this study 72 samples, maternal age 22 ≤ and ≤36, have been analyzed; maternal plasma and placental miRNAs were isolated from 18 severe preeclampsia (sPE) patients and 18 controls, respectively. Profiling of human miRNAs (1368 probe) was performed in samples with Agilent v16 microarrays for detection of the differences in miRNA expression between two groups. The results were validated by using TaqMan RT-qPCR method. The analysis indicated that 406 of these miRNAs in all placentas and 42 of these miRNAs in all maternal plasma were expressed. The relative expression analysis has shown that 12 miRNAs ( p 2-fold) in maternal plasma were differentially expressed in PE and control group. However, five miRNAs were validated by qRT-PCR. Once validated miRNAs have been searched in databases for their target genes and function, it has been shown that there are some preeclampsia related pathways as a target such as angiogenesis, cardiovascular, hypertension, placental abruption and preeclampsia disorders. Differentially expressed and validated plasma miRNAs might be used as notable biomarkers for non-invasive early diagnosis of preeclampsia and treatment of disease.

Thoracic aortic aneurysm (TAA) and thoracic aortic dissection (TAD) are aortic diseases known as 'silent killers'. While TAA is characterized by an enlargement of at least half of the normal aortic diameter, TAD is characterized by progressive pseudo‑lumen formation, which results in the gradual separation of the aortic wall layers. In the present study, a total of 28 serum samples from nine patients with TAA, nine patients with TAD and ten healthy individuals were studied. The aim of the present study was to investigate the expression profiles of hsa‑microRNA(miR)‑143‑3p and hsa‑miR‑22‑3p in TAA and TAD in order to identify candidate miRNAs that are responsible for the pathogenesis of the diseases. Following the detection of target mRNAs from candidate miRNAs by bioinformatic tools, the expression profiles of target mRNAs were analyzed. A quantitative polymerase chain reaction was performed to detect Kirsten rat sarcoma viral oncogene homolog (KRAS), mitogen‑activated protein kinase (MAPK) 7, MAPK14 and transgelin (TAGLN) mRNA expression profiles. The results of the comparison with control group demonstrated that the increase in the expression levels of hsa‑miR‑143‑3p (P=0.017) and hsa‑miR‑22 (P=0.03) candidate miRNAs were statistically significant in the TAA group, but not in the TAD group. The expression of KRAS and MAPK7 mRNAs decreased in the two groups compared with the control group. The level of expression of MAPK14 decreased in the TAD group, but increased in the TAA group compared with the control group. TAGLN mRNA expression level increased in the two groups. The statistically significant difference in the expression of hsa‑miR‑143‑3p suggests that hsa‑miR‑143‑3p may be a potential biomarker for TAA, as the expression of the target mRNAs KRAS and MAPK7 decreased and the miRNA‑mRNA association was negatively correlated. These miRNAs and their associated genes may serve important functions in TAA formation, the altered expression of which may be important in the pathogenesis of TAA.

Çevik, Nazife (Arel Author), PurposeOvarian cancer (OC) is first gynaecologic cancer that causes women death and epithelial ovarian cancer (EOC) is the most lethal ovarian cancer type. While treatment is commonly successful, some cases (10-20%) show resistance to chemotherapy which is followed by recurrence. MicroRNA (miRNA) based diagnosis methods are slightly important for recurrent ovarian cancer diagnosis. We aimed to detect novel circulating miRNAs to be used as an early diagnosis and prediction tools for recurrent EOC.MethodsIn this study, recurrent EOC serum samples and healthy control serum samples were compared for miRNA expression analysis by microarray. Microarray results were analyzed by bioinformatics tools and differentially expressed hsa-miR-1273g-3p was obtained. After microarray analysis, differentially expressed hsa-miR-1273g-3p was validated by Real-Time PCR (RT-qPCR). The relation between target genes of hsa-miR-1273g-3p and ovarian cancer were examined by Pathway Studio((R)) (v.11.4.0.8).ResultsThe expression of hsa-miR-1273g-3p was found to be significantly down-regulated by t test Bonferroni FWER corrected p2, in recurrence EOC compare with healthy controls groups. The RT-qPCR results confirmed that relative expressions of the serum hsa-miR-1273g-3p were significantly down-regulated in patients with recurrent EOC (p=0.0275). Serum hsa-miR-1273g-3p levels could discriminate patients with recurrent EOC from healthy controls, with a power area under the curve (AUC) of 0.7.ConclusionThis study suggested that hsa-miR-1273g-3p plays a significant role in regulation of related genes, which are TNF-alfa, COL1A1, MMP-2, MMP-9, with recurrent EOC outcome. hsa-miR-1273g-3p may be used as a prognostic marker for recurrent EOC after chemotherapy.

The aim of this study was to use proteomic and transcriptomic approaches to examine differences in protein and gene expression in maternal plasma between normal and preeclamptic pregnancies. Preeclampsia and control groups were compared with respect to the expression of CD34 and CD133 genes by quantitative polymerase chain reaction (qPCR) and heat shock protein (Hsp)27 and 70 by western blotting in blood samples from the pregnant women. Blood samples were obtained at gestational week 12-14 from 65 healthy pregnant women. Fetal DNA was isolated from the maternal blood and CD34 and CD133 were amplified by qPCR. Western blot analysis was used to examine the expression levels of Hsp27 and Hsp70 proteins. The analysis of CD133 by qPCR was unsuccessful in 7 women as the levels of fetal DNA were in the collected maternal blood samples were insufficient. Measurements of CD34 and CD133 were performed in 57 and 50 women, respectively. Preeclampsia developed in 6 (10.5%) of 57 women. qPCR results of 8 healthy pregnant women were used for the calibration of CD34 and CD133 levels, and the results for the remaining women were compared with the calibration values. CD34 expression was decreased in 30 (52.6%) and increased in 27 (47.4%) of 57 women. CD133 expression was decreased in 14 (28%) and increased in 36 (72%) of 50 women. CD34 expression was increased in 2 (33%) and 25 (49%) and decreased in 4 (66%) and 25 (51%) women with and without preeclampsia, respectively (P=0.467). CD133 expression was increased in 4 (66%) and 32 (72%); and decreased in 2 (33%) and 12 (28%) women with and without preeclampsia, respectively (P=0.756). Western blotting showed that the expression of Hsp27 and Hsp70 in the maternal serum of the preeclampsia group was significantly higher than that in the normal pregnancy group. CD34 and CD133 were found to be inadequate for use in the prediction of preeclampsia. However, it is noteworthy that CD133 levels were increased in 66 and 72% of women with and without preeclampsia, respectively. Hsps are expressed under various pathological conditions. These results suggest that conditions of oxidative stress increased the Hsp27 and Hsp70 protein levels.

MicroRNAs (miRNAs) are single-stranded RNA (20-23 bp) molecules which regulate expression of proteincoding gene by promoting mRNA degradation and translational inhibition in mammalians. The latest miRBase release (v20, June 2013) contains 24 521 microRNA loci from 206 species which is processed to produce 30,424 mature microRNA products. miRNAs are involved in several mechanisms such as cardiovascular, central nervous system processes and cancer. miRNAs can be produced by human placenta and are significant for placental development. Recently, regulation of miRNA expression is not exactly understood in human placenta, however many research demonstrated that oxygen tension, signaling molecules, environmental toxin and epigenetic regulation are playing critical roles in placental development. Placental miRNAs that are used for prenatal diagnosis have been detected in maternal plasma in recent years. Some complications of pregnancy such as preeclampsia which is also regulated by miRNA could be diagnosed by using miRNA techniques. In preeclamptic blood and tissue samples, upor down-regulated miRNA levels can be observed. In this study, topics such as miRNA biogenesis and regulation in pregnancy, pregnancy associated disorders are referred and future perspective of prenatal diagnosis using miRNA as biomarkers.

Recent developments in molecular genetics improved our knowledge on fetal genome and physiology. Novel scientific innovations in prenatal diagnosis have accelerated in the last decade changing our vision immensely. Data obtained from fetal genomic studies brought new insights to fetal medicine and by the advances in fetal DNA and RNA sequencing technology novel treatment strategies has evolved. Non-invasive prenatal diagnosis found ground in genetics and the results are widely studied in scientific arena. When Lo and colleges proved fetal genetic material can be extracted from maternal plasma and fetal DNA can be isolated from maternal serum, the gate to many exciting discoveries was open. Microarray technology and advances in sequencing helped fetal diagnosis as well as other areas of medicine. Today it is a very crucial prerequisite for physicians practicing prenatal diagnosis to have a profound knowledge in genetics. Prevailing practical use and application of fetal genomic tests in maternal and fetal medicine mandates obstetricians to update their knowledge in genetics. The purpose of this review is to assist physicians to understand and update their knowledge in fetal genetic testing from maternal blood, individualized prenatal counseling and advancements on the subject by sharing our experiences as İstanbul University Fetal Nucleic Acid Research Group.

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 ~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.

Genetic polymorphism is considered to be associated with human physical performance. The angio-tensin-Iconverting enzyme insertion/deletion (ACE I/D) and the α-actinin-3 gene (ACTN3) R577X polymorphisms have been widely investigated for such associations, and functional ACE I/D and ACTN3 R577X polymorphisms have been associated with sprinter performance. The aim of this study was to determine the effect of these polymorphisms on sport performance among 37 elite athletes and 37 healthy controls. The ACE II genotype was identified in 32.43% of the control group and 8.11% of elite athletes, the DD genotype in 37.84% of the control group and 51.35% of the elite athletes, and the ID genotype in 29.73% of the control group and 40.54% of the elite athletes. With regard to the ACTN3 gene, the XX geno-type, which confers an advantage for endurance activities, was identified in 10.81% of the control group and 35.14% of the elite athletes. The XX genotype was observed more frequently than the RR genotype (advantageous for sprinting), which was iden -tified in 2.70% of the control group and 10.81% of elite athletes. The RX genotype (observed in 86.48% of the control group and in 54.05% of the elite athletes) was the most common geno-type of the individuals in the present study. The study showed that ACTN3 and ACE gene polymorphisms have an effect on muscle power; however, larger studies are required.

Aim of the Study: The purpose of this study was to identify specific circulating microRNAs (miRNAs) and investigate expression level of their target genes for evaluation of pathogenesis of epithelial ovarian cancer (EOC). Materials and Methods: In this study, we have studied on EOC patients' serum and whole blood, healthy control (HC) serum, and whole blood samples. Sixteen serum samples were collected to compare miRNA expression analysis through microarray. According to microarray results, one of the dysregulated miRNA in serum, hsa-let-7d-3p, was validated by RT-qPCR for discriminate two groups. The hsa-let-7d-3p is one of the tumor suppressive let-7d family members. Let-7d is downregulated in numerous types of cancer, including ovarian cancer and directly targets various oncogenes. We analyzed the let-7d targets, which are High Mobility Group A2 (HMGA2) and (Kirsten Rat Sarcoma Viral Oncogene Homolog), as the oncogenes that are associated with EOC. The relation between target genes of hsa-let-7d-3p and EOC has been examined by Pathway Studio. Twenty serum and whole blood samples collected to analyze expression level of target genes were analyzed by real-time PCR. Results: 31 significantly dysregulated miRNAs were identified by microarray in serum. Hsa-let-7d-3p has been selected for the validation, according to P-value and dysregulated level. RT-qPCR results showed that hsa-let-7d-3p could discriminate EOC patients from HC (P = 0.0484, AUC = 0.7). Furthermore, we identified hsa-let-7d-3p's target genes (HMGA2, KRAS) by bioinformatic analysis. The expression level of genes could discriminate patients with EOC from HC, with a power area under the ROC curves (AUC) of 62 and 64.2, respectively. Conclusion: HMGA2 and KRAS could be translationally downregulated by the hsa-let-7d-3p, and the loss of hsa-let-7d-3p expression led to the progression of EOC related to the tumorigenesis, invasion, and metastasis.

Preeclampsia is associated with increased perinatal morbidity and mortality. There have been numerous efforts to determine preeclampsia biomarkers by means of biophysical, biochemical, and spectroscopic methods. In this study, the preeclampsia and control groups were compared via band component analysis and multivariate analysis using Raman spectroscopy as an alternative technique. The Raman spectra of serum samples were taken from nine preeclamptic, ten healthy pregnant women. The Band component analysis and principal component analysis-linear discriminant analysis were applied to all spectra after a sensitive preprocess step. Using linear discriminant analysis, it was found that Raman spectroscopy has a sensitivity of 78% and a specificity of 90% for the diagnosis of preeclampsia. Via the band component analysis, a significant difference in the spectra of preeclamptic patients was observed when compared to the control group. 19 Raman bands exhibited significant differences in intensity, while 11 of them decreased and eight of them increased. This difference seen in vibrational bands may be used in further studies to clarify the pathophysiology of preeclampsia.

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 RHDΨ), indicative of a maternal RHD allele. We conclude that it is necessary to analyze at least two exon regions in the RHD gene.

Hemolytic disease of the newborn (HDN) is a clinic phenomenon which occurs during pregnancy due to the Rhesus (Rh) D alloimmunization between a Rh (-) pregnant woman, who has become sensitive to RhD antigens, and her Rh (+) fetus. As a result of the attack of maternal RhD antibodies on fetal RhD antigens, fetal anemia, HDN and fetal death may occur. % 40 of Rh (-) pregnant women carry Rh (-) fetus. However, all Rh (-) pregnant women are offered anti-D Immunoglobulin (Anti-D Ig) at 28 weeks' gestation in case of fetomaternal haemorrhage, so the pregnant women carrying Rh (-) fetus are exposed to blood products unnecessarily. Although the RhD of fetus can be detected, methods used for prenatal diagnosis recently are invasive tests and they can result in abortion in a certain percentage. The discovery of circulating cell-free fetal nucleic acids in maternal plasma has opened up new possibilities for non invasive prenatal diagnosis. The aim of this study was to detect prenatal RhD by analysing the presence of the RhD gene of fetal DNA in maternal blood.Total free DNA was isolated from the blood of 19 Rh (-) pregnant women, who had RhD alloimmunization with their husbands, in the 11-14 th week of their pregnancy. The existence of a gene in isolated DNA was investigated with TaqMan prob and "Real-time PCR" method by using primers belonging to exon 7 of RhD gene.Using a quantitative real-time PCR assay, the presence of RhD gene sequences was evaluated in the serum of patients at the onset of pregnancy. We have analyzed 19 Rh (-) pregnant women. Twelve of them were Rh (-) and the rest of them were 7 Rh (+). After birth the baby's blood groups were concordant with our results.The results obtained by RhD primer were analysed. The possibility of detection of fetal RhD gene in maternal blood contributed to noninvasive prenatal diagnosis.Yeni doğanın hemolitik hastalığı, daha önce “Rhesus” (Rh) D antijenleri ile duyarlı hale gelmiş Rh (−) anne ile Rh (+) çocuğu arasındaki RhD uyumsuzluğuna bağlı olarak hamilelik sürecinde ortaya çıkan klinik bir tablodur. Maternal RhD antikorlarının fetal RhD antijenlerini yok etmesiyle, fetusta kansızlık, yenidoğanda sarılık veya fetal ölüm gerçekleşebilir. Rh (−) gebelerin yaklaşık % 40′ı Rh (−) fetus taşımaktadır. Buna rağmen tüm gebelere olası bir fetal kanama riskine karşı hamileliklerinin 28. haftasında anti D immünglobulin (Anti-D Ig) uygulanmakta ve Rh (−) fetus taşıyan gebeler gereksiz aşı ürününe maruz kalmaktadır. Doğum öncesi tanı ile fetusun RhD tayini yapılabilse de, günümüzde kullanılan doğum öncesi tanı metodları girişimsel tetkikler olup belli oranlarda gebelik kayıplarına neden olabilmektedir. Anne kanında fetusa ait nükleik asitlerin hücre dışında serbest dolaşabileceklerinin keşfi noninvazif prenatal tanı için yeni olanaklar açmıştır. Bu çalışmada da maternal kandan fetal DNA’da RhD geninin varlığına bakılarak doğum öncesi RhD tayini yapılabilmesi amaçlanmıştır.11–14. gebelik haftalarında bulunan ve eşleriyle aralarında RhD uyumsuzluğu olan 19 Rh (−) gebeden kan alınarak total serbest DNA izolasyonu yapılmıştır. İzole edilen DNA’larda RhD geninin ekson 7 bölgesine ait primerler kullanılarak, TaqMan prob yöntemi ile ve “Real-time PCR” aracılığıyla genin varlığı araştırılmıştır.“Real-time PCR” kullanılarak gebeliklerinin başında olan hastaların serumlarında RhD gen dizilerinin varlığı değerlendirildi. Analiz edilen 19 Rh (−) gebenin 12′sinin (Rh−), 7′sinin Rh (+) olduğu belirlendi. Doğum sonrasında bebeklerin kan grupları ile bizim sonuçlarımız birebir uyumlu çıkmıştır.RhD primeri ile alınan sonuçlar analiz edilmiştir. Fetüse ait RhD geninin maternal kanda belirlenebilmesi girişimsel olmayan doğum öncesi tanı için önemli bir katkı sağlamıştır.

Geranylgeranyl pyrophosphate synthase (GGPPS) is a key enzyme in carotenoid biosynthetic pathway, catalyzing the synthesis of its C20 precursor. The gene encoding GGPPS from bell pepper (Capsicum annuum) has been cloned in Schizosaccharomyces pombe and its heterologous expression analyzed in this model organism for eukaryotes. We aim to partially direct the ergosterol pathway to carotenoid biosynthesis in this yeast. This yeast is not able to synthesize carotenoids naturally, but it produces farnesyl pyrophosphate (FPP), an intermediate metabolite for both ergosterol and geranylgeranyl pyrophosphate (GGPP) biosynthesis. S. pombe transformants carrying the pepper GGPPS cDNA were named KTG. In one of the transformant (KTG1), heterologous expression of GGPPS gene has been analyzed. The GGPPS gene was shown by dot and Northern hybridization to be successfully transcribed. When water soluble proteins isolated from the transformant cells have been analyzed on SDS-denaturating gels, a ~37 kDaprotein band...

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 gre...

Basically, diagnosis inherited diseases in gestation is very important. But amplication of the progress in early stage of pregnance is the serious and great aim for rapid application of risk assesment for both and fetus. Fetal genetic tissues collected through techniques such as amniocentesis and chorionic villus sampling (CVS) for prenatal diagnosis of fetal genetic diseases. These procedures are associated with a risk of fetal loss (1%) and two week needed for cultivation (Chiu et al., 2010). Rapid methods for prenatal diagnosis of fetal chromosomal aneuploidies have been developed. This method is multicolor fluorescence in situ hybridization (FISH). FISH is very reliable but requires intact cells and it can only be used on fresh samples (Klinger et al.,1992) The other method is quantitative fluorescent polymerase chain reaction PCR anaylsis (real-time PCR) (Lo et al., 1997). Real-time PCR is a powerful tool for quantifying gene expression combining both high sensitivity and specificity with efficient signal detection (Kubista, 2008). A major advantage of real-time PCR is that it can be used to determine the amount of initial temlates (Heidi et al., 1996). In addition, by making use of a closed whereby the samples are analyzed directly in PCR reaction vessel during the amplification. Furthermore, the assay is less prone contamination (Zimmerman, 2006). Real-time PCR has found wide spread applicability in the analysis of gene expression measurement and cell-free nucleic acids in body fluids. The assay is readily amenable to automation, and by making use of the current real-time PCR 96-384 well formats. The discovery of cff DNA in maternal plasma in 1997 has opened non-invasive prenatal diagnosis (NIPD). This research area is a rapidly developing and dynamic field. NIPD using cell-free fetal DNA (and RNA) is likely to become increasingly available within the next few years. There have been a lot of reported applications, including fetal rhesus-D genotyping (Bombard et al.,2011), fetal sexing for X-linked disorders (Costa et al., 2002), paternally inherited genetic diseases, and pregnancy-associated conditions such as preeclampsia (Zhong et al., 2001). Diagnosing by using free DNA a variety of limitations due to some of the features of this resource. CffDNA analyses would be applicable only when the involved target sequence is in the fetus but absent from mother, such as paternally

Preeclampsia continues to be a mortal disease of pregnant women throughout the world. Recently, geneticists, allied with obstetricians, have opened new frontiers. MicroRNAs (miRNAs) are members of a class of small, noncoding RNA molecules. They are critical posttranscriptional regulators of gene expression. We extracted circulating miRNA from maternal plasma and quantified mir-152 and mir-210. We found up-regulated miR-210 levels as well as down-regulated mir-152 levels in preeclampsia patients.We propose that detection of increased mir-210 levels in maternal serum could be used to improve prediction methods for noninvasive prenatal diagnosis of preeclampsia.

Aim Quantitative changes of cell-free fetal DNA in maternal plasma as an indicator for impending pre-eclampsia was reported in different studies. Cell-free fetal nucleic acids can be detected in maternal circulation during pregnancy. Our aim was to determine the higher rate of fetal DNA levels in maternal blood in pre-eclampsia compared to normal pregnancies and the clinical use of real-time polymerase chain reaction (PCR) in the Turkish population as a marker. Material and Methods According to their gestational ages, the plasma levels of 30 pre-eclamptic women at 26–40 weeks of pregnancy were matched with 18 healthy pregnant women. Cell-free fetal DNA levels in maternal plasma were compared using real-time PCR technology. For the quantitative measurement of fetal DNA from maternal blood, the relative quantification PCR process was applied to all samples, using SRY and GAPDH genes. These patients were classified as pre-eclamptic and control groups and were matched according to weeks of pregnancy. Results Free fetal DNA levels of 30 pre-eclamptic patients were compared to healthy pregnant women and an average 3.06-fold increase was observed. During the second trimester, free fetal DNA levels were 1.5 times higher in pre-eclamptic patients. This increase was 3.5-fold during the third trimester. The DNA increase of pre-eclamptic patients was 4.1-fold and 3.4-fold during 29th–33rd and 34th–40th weeks, respectively. Conclusions Cell-free fetal DNA in maternal blood could be used as a marker for identifying subjects at increased risk of developing pre-eclampsia.

Hemolytic disease of the newborn is the clinical condition in which Rh blood group antigens in couples are incompatible with each other and mother is negative for the antigen, whereas father is positive. Although RHD antigen encoded by RHD gene that is localized on chromosome 1 determines person's Rh genotyping, this incompatibility can lead to delivery as anemia, jaundiced, or dead in mother's uterus. In recent years, improvements have occurred in the prenatal diagnosis of Rh incompatibility. Quantitative real-time polymerase chain reaction (Real-time PCR) has been improved and determining rapidly, reliably, and sensitively has been possible. In this study, the determination of RHD genotyping was investigated using fetal DNA obtained from amniotic fluid and SYBR Green I and TaqMan probe methods were compared, and reliability in prenatal diagnosis of these methods was determined. We studied 35 pregnant women in the second trimester of pregnancy. "SYBR Green I" and "TaqMan" probes results for RHD gene of genomic DNA extracted from total 35 different amniotic fluid samples acquired from 10 RHD (-) and 25 pregnant women randomly were analyzed. DNA extracted from amniotic fluid was analyzed for RHD gene with real-time PCR and the results were then compared with the RHD fetal genotype determined on RHD phenotype of the red blood cells of the infants at birth. The results of RHD TaqMan probes PCR analysis of amniotic fluid DNA were completely concordant with the fetal blood group analysis after birth. Real-time PCR using the TaqMan probes has proven to be more sensitive, accurate, and specific for RHD gene than SYBR Green I method.