Genomisches Imprinting und Imprintingerkrankungen beim Menschen

Imprinting ist ein epigenetischer Prozess, über den die monoallelische Expression gewährleistet wird. Imprintingfehler führen zu Entwicklungsstörungen und einer veränderten Genexpression, was zu Imprintingerkrankungen führt. Das Prader-Willi Syndrom (PWS) wird durch den Funktionsverlust paternal exprimierter Gene in der 15q11q13 Region hervorgerufen, wobei allerdings nicht klar ist, welche Rolle die einzelnen Gene bei der phänotypischen Ausprägung des PWS spielen. In dieser Arbeit konnte durch eine Genotyp/Phänotyp Analyse von zwei Patienten mit PWS mit einer atypischen Deletion in 15q11q13 und einer Patientin mit einer unbalancierten Translokation t(X;15)(q28;q11.2) gezeigt werden, dass die paternal exprimierten Gene MKRN3, MAGEL2 und NDN keine wesentliche Bedeutung bei der Entstehung des PWS haben. Durch eine Kombination von Methylierungs-, Segregations- und Gendosisanalysen bei sieben Patienten konnte bestätigt werden, dass Imprintingfehler und Deletionen in der DLK1/MEG3 Region zu einem Phänotyp führen, der vergleichbar mit dem Phänotyp ist, der ursprünglich bei dem Vorliegen einer maternalen uniparentalen Disomie 14 beschrieben wurde. Demnach scheint der Funktionsverlust der paternal exprimierten Gene DLK1 und RTL2 für den upd(14)mat Phänotyp verantwortlich zu sein. In einem weiteren Teil dieser Arbeit wurde untersucht, ob Kinder, die durch ICSI gezeugt wurden und ein niedriges Geburtsgewicht hatten, abnormale Methylierungsmuster an geprägten Loci aufweisen. Dazu wurden sechs geprägten Loci bei 19 ICSI-Kindern und 29 gleichaltrigen Kontroll-Kindern (natürlich gezeugt; normales Geburtsgewicht) untersucht. Unter 19 ICSI-Kindern zeigte ein ICSI-Kind Methylierungsveränderungen am KCNQ1OT1 und am MEST Locus. Im Rahmen dieser Arbeit wurde zudem mittels ausgedehnten Methylierungsanalysen bei einem Patienten eine generalisierte Imprintingstörung nachgewiesen. Er zeigte eine Hypomethylierung sowohl an maternal als auch an paternal methylierten Loci. Die aberranten Methylierungsmuster liegen als somatisches Mosaik vor. Bislang wurde kein weiterer Fall mit einer Hypomethylierung an allen bekannten geprägten Loci beschrieben. Ausgehend von einer genomweiten Methylierungsanalyse dieses Patienten konnte gezeigt werden, dass im Intron 2 des RB1 Gens ein elternspezifisch methyliertes CpG island lokalisiert ist, das einen alternativen RB1 Promotor beinhaltet. Das neu identifizierte, alternative RB1 Transkript wird ausschließlich vom paternalen Allel exprimiert. Weitere Untersuchungen sind notwendig, um die Funktion dieses Transkripts und dessen klinische Relevanz zu klären

Fetal conotruncal heart anomalies: Is fourchamber view sufficient in the prenatal screening?

Amaç: Bu çalışmada hastanemiz perinatoloji ünitesinde konotrunkalkalp anomalisi tanısı alan olguların kayıtları incelenerek prenataldönemde konotrunkal kalp anomalisi tanısına yönlendiren faktörlerindeğerlendirilmesi ve prenatal konotrunkal kalp anomalisitanısı ile ilgili farkındalığın artırılması amaçlandı.Yöntem: Ocak 2015 – Aralık 2016 tarihleri arasında Kanuni SultanSüleyman Eğitim ve Araştırma Hastanesi Perinatoloji Ünitesindekonotrunkal kalp anomalisi tanısı alan olguların yönlendirilmenedenleri, eşlik eden kalp dışı anomali ve kromozom anomalisivarlığı, olguların perinatal sonuçları ve postnatal dönemde tanı-nın doğrulanma başarısı değerlendirildi.Bulgular: Tüm konjenital kalp anomalilerinin içinde konotrunkalkalp anomalisi sıklığı %20.4 idi. Çalışmaya dahil edilen 101 olgunun37’sinde (%36.6) gebelik terminasyonu gerçekleştirildi. ‹ntrauterinfetal ölüm 5 (%5) olguda gözlendi. Olguların %26.7’sindekromozom anomalisi ve %34.7’sinde kalp dışı ek yapısal anomalisaptandı. Canlı doğan 59 (%58.4) olgunun 52’sinde (%88.1) prenataltanı doğrulandı. Olguların yalnızca %27.7’sinde dört oda görünümününanormal olduğu saptandı.Sonuç: Dört oda görünümünün konotrunkal kalp anomalilerindesıklıkla normal olması nedeni ile bu anomalilerin prenatal tanı sıklığının artırılabilmesi için temel fetal kardiyak tarama programlarında üç damar ve üç damar trakea kesitleri rutin olarak görüntülenmelidir.Objective: In this study, we aimed to assess the factors leading to the diagnosis of conotruncal heart anomaly in the prenatal period by reviewing the records of the cases which were diagnosed with the conotruncal heart anomaly in our perinatology unit, and to raise the awareness of the diagnosis of prenatal conotruncal heart anomaly. Methods: The referral reasons, the presence of concomitant non-cardiac anomaly and chromosomal anomaly, perinatal outcomes of the cases and the confirmation success of the diagnosis at postnatal period of the cases which were diagnosed with the conotruncal hearth anomaly at the Perinatology Unit of Kanuni Sultan Süleyman Training and Research Hospital between January 2015 and December 2016 were evaluated. Results: Among all congenital cardiac anomalies, the incidence of conotruncal heart anomaly was 20.4%. The termination of pregnancy was performed in 37 (36.6%) of 101 cases included in the study. Intrauterine fetal death was observed in 5 (5%) cases. Chromosomal anomaly and non-cardiac additional structural anomaly were found in 26.7% and 34.7% of the cases, respectively. Prenatal diagnosis was confirmed in 52 (88.1%) of 59 (58.4%) cases which born alive. It was found that four-chamber view was abnormal in only 27.7% of the cases. Conclusion: Three vessels (3V) and three vessels trachea (3VT) views should be displayed routinely in basic fetal cardiac screening in order to increase the prenatal diagnosis frequency of these anomalies as four-chamber view is mostly normal in conotruncal heart anomaly

Применение микробиологических методов для повышения нефтеотдачи на примере нефтяного месторождения Мухто (Сахалинская область)

Подбор и применение микробиологического метода увеличения нефтеотдачи на месторождении РН-Сахалинморнефтегаз.Актуальность этого метода заключается в том, что этот метод позволяет извлекать трудноизвлекаемые запасы нефти, которые увеличиваются с каждым годом. В этой работе выбирается наиболее эффективный микробный агент, и рассчитывается рентабельность этого нововведения для компании.Selection and application of the microbiological method of increasing oil recovery at the RN-Sakhalinmorneftegaz.The relevance of this method lies in the fact that this method allows you to extract hard-to-recover oil reserves, which increase every year. In this paper, the most effective microbial agent is selected, and the profitability of this innovation for the company is calculated

The Human Retinoblastoma Gene Is Imprinted

Genomic imprinting is an epigenetic process leading to parent-of-origin–specific DNA methylation and gene expression. To date, ∼60 imprinted human genes are known. Based on genome-wide methylation analysis of a patient with multiple imprinting defects, we have identified a differentially methylated CpG island in intron 2 of the retinoblastoma (RB1) gene on chromosome 13. The CpG island is part of a 5′-truncated, processed pseudogene derived from the KIAA0649 gene on chromosome 9 and corresponds to two small CpG islands in the open reading frame of the ancestral gene. It is methylated on the maternal chromosome 13 and acts as a weak promoter for an alternative RB1 transcript on the paternal chromosome 13. In four other KIAA0649 pseudogene copies, which are located on chromosome 22, the two CpG islands have deteriorated and the CpG dinucleotides are fully methylated. By analysing allelic RB1 transcript levels in blood cells, as well as in hypermethylated and 5-aza-2′-deoxycytidine–treated lymphoblastoid cells, we have found that differential methylation of the CpG island skews RB1 gene expression in favor of the maternal allele. Thus, RB1 is imprinted in the same direction as CDKN1C, which operates upstream of RB1. The imprinting of two components of the same pathway indicates that there has been strong evolutionary selection for maternal inhibition of cell proliferation

Clinical relevance of postzygotic mosaicism in Cornelia de Lange syndrome and purifying selection of NIPBL variants in blood

Postzygotic mosaicism (PZM) in NIPBL is a strong source of causality for Cornelia de Lange syndrome (CdLS) that can have major clinical implications. Here, we further delineate the role of somatic mosaicism in CdLS by describing a series of 11 unreported patients with mosaic disease-causing variants in NIPBL and performing a retrospective cohort study from a Spanish CdLS diagnostic center. By reviewing the literature and combining our findings with previously published data, we demonstrate a negative selection against somatic deleterious NIPBL variants in blood. Furthermore, the analysis of all reported cases indicates an unusual high prevalence of mosaicism in CdLS, occurring in 13.1% of patients with a positive molecular diagnosis. It is worth noting that most of the affected individuals with mosaicism have a clinical phenotype at least as severe as those with constitutive pathogenic variants. However, the type of genetic change does not vary between germline and somatic events and, even in the presence of mosaicism, missense substitutions are located preferentially within the HEAT repeat domain of NIPBL. In conclusion, the high prevalence of mosaicism in CdLS as well as the disparity in tissue distribution provide a novel orientation for the clinical management and genetic counselling of families.Spanish Ministry of Health-ISCIII Fondo de Investigación Sanitaria (FIS) [Ref. PI19/01860, to F.J.R. and J.P.]; Diputación General de Aragón-FEDER: European Social Fund [Grupo de Referencia B32_17R / B32_20R, to J.P.]. A.L-P is supported by a “Juan de la Cierva-Incorporación” postdoctoral grant from MICIU (Spanish Ministry of Science and Universities

Generation of heterozygous and homozygous hESC H9 sublines carrying inactivating mutations in RB1

Inactivation of the tumor suppressor gene RB1 is causal for development of retinoblastoma, a tumor of the neural retina arising in children under the age of five. In addition, secondary RB1 mutations are found in many other tumor types. To investigate retinoblastoma formation in vitro, stem cells with inactivated RB1 can be differentiated into neural retina. To enable such studies, two sublines of hESC line H9 carrying mutations in RB1 exon 3 in heterozygous or homozygous state were generated and characterized. Homozygous mutation led to loss of RB1 protein expression.Resource tableUnlabelled TableUnique stem cell lines identifierWAe009-A-12WAe009-A-13Alternative names of stem cell linesC7 (homozygous deletion, WAe009-A-12)G12LS (heterozygous deletion, WAe009-A-13)InstitutionUniversity Hospital Essen, University Duisburg-Essen, Essen, GermanyContact information of distributorDr. Laura Steenpass, [email protected]. Deniz Kanber, [email protected] of cell linesESCOriginHumanCell SourceHuman ESC line H9 purchased from WiCellClonalityClonalMethod of reprogrammingN/AMultiline rationaleclones selected for deletion in heterozygous and homozygous stateGene modificationYESType of modificationIndels in RB1 exon 3Associated diseaseRetinoblastomaGene/locusRB1, chromosome 13q14.2Method of modificationCRISPR/Cas9 nucleaseName of transgene or resistanceN/AInducible/constitutive systemN/ADate archived/stock dateC7 12.05.2018G12LS 12.05.2018Cell line repository/bankN/AEthical approvalApproval obtained from the Robert-Koch Institute, Berlin, Germany (Az.3.04.02/0101) and from the local Ethical Review Board University Duisburg-Essen (16–7215-BO

CHRONIC INFLAMMATION MARKERS hs-CRP, sICAM-1, sVCAM-1 AND sE-SELECTIN LEVELS IN PATIENTS WITH POLYCYSTIC OVARY SYNDROME

Objective: Patients with polycystic ovary syndrome have many risk factors associated with development of cardiovascular disease development. Presence of low grade chronic inflammation has been indicated as a risk factor for cardiovascular disease. We investigated the levels of hs-CRP, sICAM-1, sVCAM-1 and sE-Selektin as markers of chronic inflammation

Comparison of fetal cardiac functions between small-for-gestational age fetuses and late-onset growth-restricted fetuses

Background: This study aimed to investigate fetal cardiac functions by spectral tissue Doppler imaging (s-TDI) in pregnancies complicated with late-onset fetal growth restriction (LO-FGR) and small-for-gestational age (SGA). Methods: Forty pregnancies complicated with late-onset FCR and 40 pregnancies complicated with SGA between the 34th and 37th weeks of gestation were enrolled in this study. Forty gestational age-matched pregnant women with no obstetrics complication were randomly selected as a control group. Small fetuses were classified as fetal growth restriction or SGA according to estimated fetal weight (EFW), umbilical artery pulsatility index (PI), cerebroplacental ratio (CPR) and uterine artery PI. s-TDI measurements were obtained at the right atrioventricular valve annulus. Results: SGA and LO-FGR fetuses had significantly lower A' and S' values, and higher E'/A' ratio than the control group (P < 0.001). In comparison to controls, significantly prolonged isovolumetric contraction time (ICT') and isovolumetric relaxation time (IRT') and, significantly shortened ejection time (ET') were observed in fetuses with SGA and LO-FGR. Increased myocardial performance index (MPI') values were also found in fetuses with SGA and LO-FGR compared to controls. Conclusion: The signs of cardiac dysfunction were observed both in fetuses with SGA and LO-FGR. The fetal cardiac function assessment with s-TDI could be a valuable method in the diagnosis of true growth restricted fetuses and in the management of these fetuses

Fetal bronchopulmonary malformations: Prenatal diagnosis and perinatal outcomes

Amaç: Bu çalışmada prenatal dönemde konjenital pulmoner hava yolu malformasyonu ve bronkopulmoner sekestrasyon tanısı alan olguların klinik özelliklerinin, prognozu etkileyen faktörlerin ve perinatal sonuçlarının değerlendirilmesi amaçlandı. Yöntem: Ocak 2013-Aralık 2016 tarihleri arasında hastanemiz perinatoloji ünitesinde konjenital pulmoner hava yolu malformasyonu ve bronkopulmoner sekestrasyon tanısı alan 59 olgunun kayıtları incelendi. Torakal kitlenin özellikleri ve doğal seyri ile olguların yönetimleri ve perinatal sonuçları incelendi. Bulgular: Otuz yedi olgu (%71.2) konjenital pulmoner hava yolu malformasyonu ve 15 olgu (%28.8) bronkopulmoner sekestrasyon tanısı aldı. Olguların %17.3’de eşlik eden yapısal anomali saptanırken, eşlik eden yapısal anomalisi olan olguların 4’ünde (%7.7) kromozom anomalisi mevcuttu. Olguların %75’inde canlı doğum gerçekleşti. Yenidoğan döneminde 23 olguda (%59) bronkopulmoner malformasyon varlığı radyolojik olarak gösterildi ve bu olguların 18’i (%78.3) yaşamın ilk bir yılında opere edildi. Sonuç: Eşlik eden anomali ve hidrops yokluğunda fetal bronkopulmoner malformasyon olgularının prenatal dönemde oldukça iyi seyrettiği ve özellikle spontan regresyon izlenen olguların postnatal dönemde operasyon gereksinimi olmadan sorunsuz yaşamlarını sürdürdüklerini saptadık. Fetal bronkopulmoner malformasyonlar ve diğer torakal anomalilerin prenatal tanı sıklığının artırılabilmesi amacıyla temel fetal anatomik tarama sırasında toraks transvers kesitlerden görüntülenmeli ve klinik şüphe varlığında çoklu sonografik planlar kullanılarak ayırıcı tanı yapılmalıdır.Objective: To evaluate the clinical features, prognostic factors and perinatal outcomes of fetuses with the diagnosis of congenital pulmonary airway malformation and bronchopulmonary sequestration. Method: The medical records of 59 fetuses with the diagnosis of congenital pulmonary airway malformation and bronchopulmonary sequestration were retrospectively analyzed. The characteristics and natural history of the thoracic mass, and the perinatal outcomes of cases were evaluated. Results: While 37 cases (71.2%) were diagnosed as congenital pulmonary airway malformation, 15 cases (28.8%) were diagnosed as bronchopulmonary sequestration. Associated structural anomalies were detected in 17.3 percent of cases and chromosomal anomalies were present in 7.7 percent of cases with associated anomalies. Live birth occured in 75 percent of cases. The presence of bronchopulmonary malformation was shown in 59 percent of cases in postnatal period, and 78.3 percent of these cases were operated in the first year of life. Conclusion: We observed that fetuses with the diagnosis of bronchopulmonary malformation had a good prognosis in the absence of associated anomaly and fetal hydrops. It is also observed that the cases with spontaneous regression of the thoracic mass did not need an operation. Thoracic transverse sections should be evaluated during basic fetal anatomic screening in order to increase the frequency of prenatal diagnosis of fetal thoracic anomalies