Effizienz der Gentherapie mit adenoviralen Vektoren bei Ornithintranscarbamylse defizienten Mäusen in Abhängigkeit von OTC-Leadersequenz

Die Effizienz einer somatischen Gentherapie des Ornithintranscarbamylase-Mangels mit adenoviralen Vektoren wird am Modell der spfash-Maus untersucht. Spfash-Mäuse werden mit Vektoren transfiziert, die das komplette OTC-Transgen der Maus, des Menschen oder Hybride mit vertauschten Leadersequenzen enthalten. Auf ultradünnen Gefrierschnitten des Lebergewebes von Kontrollmäusen, unbehandelten und transfizierten spfash-Mäusen wird die intrazelluläre Verteilung von OTC, CPS I und ATPase(c) immunelektronenmikroskopisch dargestellt und morphometrisch quantifiziert. Die Ergebnisse dieser In-vivo-Studie zeigen, dass die transgene OTC sehr effizient in die Mitochondrien von spfash-Hepatozyten importiert wird und die Voraussetzung für eine metabolische Korrektur des OTC-Mangels gegeben ist. Die mitochondriale Importkapazität der transgenen OTC und damit der gentherapeutische Erfolg hängen jedoch entscheidend von spezies-spezifischen Unterschieden in der Leadersequenz ab. The efficiency of somatic gene therapy of OTC deficiency with adenoviral vectors is examined in the spfash mouse model. Spfash mice are transfected with vectors containing the complete murine OTC transgene, the complete human OTC transgene or hybrids with exchanged leadersequences. On ultrathin frozen sections of liver tissue of control mice, untreated and transfected spfash mice the intracellular distribution of OTC, CPS I and ATPase(c) is shown by immune electronmicroscopy and is morphometrically quantified. The results of this in-vivo study reveal a very efficient import of transgenic OTC into mitochondria of spfash hepatocytes, so that the precondition for a metabolic correction of OTC deficiency is fulfilled. However, the capacity of mitochondrial import of transgenic OTC and, therefore, of gene therapeutic success strongly depend on species specific differences in the leadersequence

Preterm prelabor rupture of membranes and outcome of very-low-birth-weight infants in the German Neonatal Network.

OBJECTIVE:It was the aim of our study to evaluate the independent effect of preterm prelabor rupture of membranes (PPROM) as a cause of preterm delivery on mortality during primary hospital stay and significant morbidities in very-low-birth-weight (VLBW) infants < 32 weeks of gestation. DESIGN:Observational, epidemiological study design. SETTING:Population-based cohort, German Neonatal Network (GNN). POPULATION:6102 VLBW infants were enrolled in GNN from 2009-2012, n=4120 fulfilled criteria for primary analysis (< 32 gestational weeks, no pre-eclampsia, HELLP (highly elevated liver enzymes and low platelets syndrome) or placental abruption as cause of preterm birth). METHODS:Multivariable logistic regression analyses included PPROM as potential risk factors for adverse outcomes and well established items such as gestational age in weeks, birth weight, antenatal steroids, center, inborn delivery, multiple birth, gender and being small-for-gestational-age. RESULTS:PPROM as cause of preterm delivery had no independent effect on the risk of early-onset sepsis, clinical sepsis and blood-culture proven sepsis, while gestational age proved to be the most important contributor to sepsis risk. The diagnosis of PPROM was associated with an increased risk for bronchopulmonary dysplasia (BPD; OR: 1.25, 95% CI: 1.02-1.55, p=0.03) but not with other major outcomes. CONCLUSIONS:The diagnosis of PPROM per se is not associated with adverse outcome in VLBW infants < 32 weeks apart from a moderately increased risk for BPD. Randomized controlled trials with primary neonatal outcomes are needed to determine which subgroup of VLBW infants benefit from expectant or intentional management of PPROM

Clinical characteristics of VLBW cohort categorized according to PPROM.

<p>p-values are derived from Fisher´s exact test or Mann-Whitney-U-test if indicated (*)</p><p>Clinical characteristics of VLBW cohort categorized according to PPROM.</p

Treatment differences of VLBW cohort categorized according to PPROM.

<p>p-values are derived from Fisher´s exact test or Mann-Whitney-U-test if indicated (*)</p><p>Treatment differences of VLBW cohort categorized according to PPROM.</p

Short-term outcome in infants born after PPROM categorized according to diagnosis of amniotic infection syndrome (AIS).

<p>p-values are derived from Fisher´s exact test or Mann-Whitney-U-test if indicated (*)</p><p>Short-term outcome in infants born after PPROM categorized according to diagnosis of amniotic infection syndrome (AIS).</p

Short-term outcomes categorized according to PPROM.

<p>p-values are derived from Fisher´s exact test or Mann-Whitney-U-test if indicated (*)</p><p>Short-term outcomes categorized according to PPROM.</p

Multivariable logistic regression analysis for mortality and morbidities including well known-risk factors.

<p>Multivariable logistic regression analysis for mortality and morbidities including well known-risk factors.</p