Cellular and viral microRNAs (miRNAs) are involved in many different processes of key importance and more than 10,000 miRNAs have been identified so far. In general, relatively little is known about their biological functions in mammalian cells because their phenotypic effects are often mild and many of their targets still await identification. The recent discovery that Epstein-Barr virus (EBV) and other herpesviruses produce their own, barely conserved sets of miRNAs suggests that these viruses usurp the host RNA silencing machinery to their advantage in contrast to the antiviral roles of RNA silencing in plants and insects. We have systematically introduced mutations in EBV's precursor miRNA transcripts to prevent their subsequent processing into mature viral miRNAs. Phenotypic analyses of these mutant derivatives of EBV revealed that the viral miRNAs of the BHRF1 locus inhibit apoptosis and favor cell cycle progression and proliferation during the early phase of infected human primary B cells. Our findings also indicate that EBV's miRNAs are not needed to control the exit from latency. The phenotypes of viral miRNAs uncovered by this genetic analysis indicate that they contribute to EBV-associated cellular transformation rather than regulate viral genes of EBV's lytic phase.

Non-invasive prenatal testing (NIPT) by random massively parallel sequencing of maternal plasma DNA for multiple pregnancies is a promising new option for prenatal care since conventional non-invasive screening for fetal trisomies 21, 18 and 13 has limitations and invasive diagnostic methods bear a higher risk for procedure related fetal losses in the case of multiple gestations compared to singletons. In this study, in a retrospective blinded analysis of stored twin samples, all 16 samples have been determined correctly, with four trisomy 21 positive and 12 trisomy negative samples. In the prospective part of the study, 40 blood samples from women with multiple pregnancies have been analyzed (two triplets and 38 twins), with two correctly identified trisomy 21 cases, confirmed by karyotyping. The remaining 38 samples, including the two triplet pregnancies, had trisomy negative results. However, NIPT is also prone to quality issues in case of multiple gestations: the minimum total amount of cell-free fetal DNA must be higher to reach a comparable sensitivity and vanishing twins may cause results that do not represent the genetics of the living sibling, as described in two case reports.

The Epstein-Barr virus (EBV) latency III program imposed by EBNA2 and LMP1 is directly responsible for immortalization of B cells in vitro and is thought to mediate most immunodeficiency-related (45), reflecting the transforming capacity of the virus and the destruction of the T-cell compartment of the host by the immunodeficiency (27). Thus, it is usually conceded that LCLs represent an in vitro model of PTLDs. EBV is also associated with various cancers, including BL, Hodgkin's lymphomas, T-cell lymphomas, and nasopharyngeal carcinomas.The EBV genome persists in the host cell in an episomal form. Three latency viral programs have been described for this virus both in vitro and in vivo. Two small nonpolyadenylated RNAs (EBER1 and EBER2) and a large transcription unit called BART (BamHI-A rightward transcript) or CST (complementary strand transcript), giving rise to a number of microRNAs, are expressed in all forms of latency. Latency I is characterized by the expression of the viral protein EBNA1

Late-breaking abstract presentation during the plenary session at the ISPD conference, Tuesday, 14 July 2015.Funding sources: This work was funded by Life Codexx AG. Conflicts of interest: WH is an employee and shareholder of LifeCodexx AG. SG, US and JB are employees of LifeCodexx AG. CvK is a member of the Supervisory Board of LifeCodexx AG and shareholder of LifeCodexx AG. The other authors declare no conflict of interest.Non-invasive prenatal testing (NIPT) for common fetal trisomies has been radically changing prenatal diagnosis.1 Several hundreds of thousands of tests are being performed worldwide every year. Until today the fetal fraction in cell-free maternal plasma DNA is the most critical determinant for a successful NIPT analysis which is affected by certain parameters such as maternal weight 2 or the presence of an aneuploidy. 3 No other pregnancy-related factors, crucial for a reliable NIPT analysis, have been identified yet. We provide the first report describing an effect of drugs on circulating plasma nucleic acids and subsequent NIPT results. Low molecular weight heparin (LMWH) is widely used as anticoagulants in placenta-mediated pregnancy complication. 4 We show that in pregnant women on LMWH medication the cell-free plasma DNA contains a higher proportion of small DNA fragments featuring an unusually high guanosin-cytosin (GC) content than in unaffected women. This apparently biases NIPT results so that they cannot be interpreted correctly and even may provide false results. For such cases our report provides guidance that blood sampling for NIPT should ideally occur right before the next application of LMWH when the LMWH level is lowest in blood. Within three months of the year 2014, we performed a total of 1614 routine NIPT analyses (Method in the Supplementary Appendix). In 12 samples (0.74%), results could not be interpreted correctly due to an elevated GC content of higher than 44.0% in comparison to the average GC content of 42.0% estimated for the majority of analyzed plasma samples (data not shown). In correlation with the elevated GC content the z-score of chromosome 18 increased whereas the z-scores for chromosome 13 and chromosome 21 decreased. Thus, not assessing the GC content of a sample as a quality criterion of NIPT leads to falsepositive results for trisomy 18 or false negative results for trisomy 13 or trisomy 21. Repeat analysis on the routine backup blood samples (taken at the same time of the first blood sample) failed for the same reason. The clinical records of the patients with failed analyzed samples revealed that nine of these twelve women were on LMWH prophylaxis. The indications for the LMWH medication were thrombosis prophylaxis, protein C deficiency, risk of pulmonary embolism and repeated miscarriages. For five of these women (Table 1 and Table S1 in the Supplementary Appendix), an additional blood sampling was carried out right before the next pending LMWH injection, when the plasma level from the previous injection was lowest. Then, routine NIPT of these pre-LM...

Our newly established non-invasive prenatal test allows detection of fetal trisomies 13, 18, and 21 with high accuracy in a population in Germany and Switzerland.

SummaryThe particular epidemiological features of Burkitt lymphoma (BL) in Tropical Africa, first described by Denis Burkitt in 1958, initiated the search for a virus that induces malignant B cell lymphomas in humans and is transmitted by arthropods. The herpes virus (Epstein-Barr virus, EBV) discovered by Epstein and collaborators in cell lines established from BL biopsies fulfilled some of these predictions. It drives primary B cells into unlimited proliferation, induces malignant B cell lymphomas in immunocompromised individuals (post-transplant lymphoproliferative disease, PTLD) in vivo, and footprints of the virus are generally detected in African BL biopsies supporting a causative role of the virus in the pathogenesis of BL. The virus is, however, not transmitted by arthropods and is spread ubiquitously amongst the human population through saliva. Furthermore, BL and EBV-induced PTLD are now recognized as pathogenetically distinct entities: BL involves MYC-immunoglobulin translocations in contrast to PTLD, and different patterns of viral genes are expressed in both diseases. Viral gene products expressed in BL are assumed to contribute to inhibition of apoptosis, although their precise mechanism of action is not fully understood. In the future, next generation sequencing is expected to shed more light on the contribution of EBV to the pathogenesis of BL.

The authors wish to make the following corrections to this paper [1]: On page 683 at the end of Section 3.2. lines 13–14, the word “no” is missing. The correct sentence should be: “There has been no evidence of false-negative NIPT results so far in the pregnancies included in this study.”[...]

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