Recombinant luciferase-expressing murine gammaherpesvirus 68 as a tool for rapid antiviral screening.

Murine gammaherpesvirus 68 (MHV-68) provides a valuable tool to screen novel therapeutic strategies against oncogenic gammaherpesviruses. The development and characterization of antiviral agents usually depend on appropriate screening assays. The aim of this study was to develop rapid and sensitive method for testing antiviral compounds against gammaherpesviruses. For this purpose, a recombinant MHV-68 expressing firefly luciferase (MHV-68/LUC) was constructed. The conditions for MHV-68/LUC infection in Vero cells suitable for novel antiviral screening assay in 96-well plate format were then optimized. The sensitivity of MHV-68/LUC to acyclovir (ACV) and ganciclovir (GCV) was measured by the optimized luciferase activity reduction assay. The 50% inhibition concentration (IC50) values for ACV and GCV were comparable to those determined by conventional plaque reduction assay. Therefore, the luciferase activity reduction assay can efficiently replace the plaque reduction assay. The great advantages of novel assay are represented by the significant reduction in assay time and rapid and objective measurement of the assay. In order to evaluate whether the luciferase activity reduction assay could be used as a screening system for novel antivirals, newly synthesized quinolone/quinoline derivatives were tested for their effects on the replication of MHV-68/LUC in vitro. The compound 2-(1-(b-D-Xylopyranosyl)-1,2,3-triazol-4-yl)-3,4-dibenzyloxy-quinoline showed significant antiviral activity and its IC50 against MHV-68/LUC was estimated to be 1,76 µg/ml. However, this compound was not suitable for in vivo testing due to its narrow selectivity index (SI = 11). Keywords: MHV-68; antiviral screening; luciferase; quinolone/quinoline derivatives.

[Pregnancy and delivery in a patient with pure 46,XY karyotype. Summary of actual knowledge about XY women]. / Tehotenství a porod u pacientky s cistým karyotypem 46,XY Souhrn dosavadních poznatku o XY zenách.

TYPE OF STUDY: Summary review and a case report. SETTINGS: GEST IVF, Centre of Reproductive Medicine, Prague. INTRODUCTION: In scientific literature there two syndrome have been described in the presence of pure 46,XY karyotype when an individual is phenotypically and psychosexually identified as a woman. Androgen insensitivity syndrome (AIS) and pure gonadal dysgenesis XY (GD XY, Swyer syndrome). Thanks to the presence of a uterus in Swyer syndrome we can treat this type of sterility with donated oocytes. METHOD: The paper describes both syndromes from prenatal, genetical, endocrinological, oncological, reproductive and perinatological points of view. A case study concerning a patient with pure gonadal dysgenesis XY, who successfully became pregnant through a donated oocytes programme, is also described. The pregnancy progressed physiologically, and a healthy boy, 3820g/52cm, was delivered in term by ceasarean section. DISCUSSION: In world scientific literature at least fifteen successful pregnancies with pure gonadal dysgenesis XY have been described. In spite of the expectation of diminished uterine capacity, children are born to term with a normal delivery weight. CONCLUSION: This article should be considered as a summary of all actual knowledge about these patients. This article should be available and usefull for clinicians who come across XY females. The case study provides evidence that even an individual with male genetic gender can be pregnant and deliver a healthy child.

[Monochorionic biamniotic twins with a common yolk sac in the first trimester ultrasound scan - is there a higher risk of a congenital defect?]. / Monochoriální biamniální gemini se spolecným zloutkovým váckem pri UZ vysetrení v I. trimestru -je zde zvýsené riziko vrozené vývojové vady?

TYPE OF STUDY: Case report. SETTING: GEST IVF, Centre of Reproductive Medicine, Prague. METHODS: The case report describes pregnancy after IVF + ET. The first trimester ultrasound scan showed monochorionic biamniotic twins with one common yolk sac (YS) only, which is possible in biamniotic twins, but very rare. Identical congenital defects such as anal, coccygeal atresia, and others, were diagnosed prenatally and postnatally in both twins. CONCLUSION: Yolk sac number in monochorionic twins can play an important role in embryogenesi. Any possible relation with certain congenital defects has not yet been described. It is necessary to describe more cases.