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Resume- Le cycle de replication du virus de l'ecthyma contagieux a ete identifie dans des etudes in vitro et un modele hypothetique a ete developpe. Pendant la premiere phase, qui dure a peu pres 5 heures, le virus penetre la cellule par le biais d'un processus de phagocytose, et perd ses enveloppes. La phase d'eclipse, pendant laquelle le virus est apparemment integrea l'ADN de l'hote, dure environ 8 a 10 heures. Pendant la phase finale, les virions se developpent dans des zones biens definies du viroplasme a partir desquelles les viriojns matures vont migrer jusqu'aux bords de la cellule. La, ils sortent soit par exocytose, soit a l'interieur de projections microvilleuses qui sont pincees a leur base, soit encore par desintegration de la cellule hote. [Onwuka, S.K., Jenkinson, D. Mc, Inglis, L., Pow, I., GRAY, E.W., Reid, H.W. Ultrastructural studies of orf virus infection and replication in fetal lamb fibrocytes (Etudes ultrasturcturales de l'infection par le virus de l'ecthyma contagieux et de sa replication dans les fibrocytes de foetus d'agneau). Resumen- Se identifico el ciclo de replicacion del virus del ectima contagioso en estudios temporales in vitro y se desarrolo un posible modelo experimental. Durante la primera fase, que dura unas 5 h, el virus penetra en la celulas por fagocitosis y se libera de la cubierta. La fase de "eclipse", con el virus presentandose como hebras de DNA, dura aproximadamente de 8 a 10 h. En la fase final los viriones se desarrollan dentro de zonas bien definidas en el viroplasma desde las cuales los viriones maduros migran hasta los limites celulares. A partir de alii parecen salir por exocitosis o en proyecciones de microvellosidades "pinzadas" hacia el exterior; tambien pueden ser liberados como consecuencia de la desintegracion de la celula huesped. [Onwuka, S.K., Jenkinson, D. Mc, Inglis, L., Pow, I., GRAY, E.W., Reid, H.W. Ultrastructural studies of orf virus infection and replication in fetal lamb fibrocytes (Estudios ultraestructurales de la infeccion por el virus del ectima contagioso y replicacion en fibrocitos fetales de carnero). Abstract- The cycle of replication of orf virus was identified in temporal in vitro studies and a putative model was developed. During the first phase, which lasts about 5 h, the virus enters the cells by a phagocytic process and uncoats. The "eclipse" phase, with the virus apparently present as strands of DNA, lasts for approximately the next 8-10 h. In the final phase virions develop within well-defined zones of viroplasm from which mature virions migrate to the margins of the cell. There they apparently exit either by exocytosis or within microvillous projections which are "pinched off"; they can also be released by disintegration of the host cell.

BACKGROUND: Maternal transplacental administration of sildenafil is being considered for a variety of fetal conditions. Clinical translation also requires evaluation of fetal safety in a higher species, such as the fetal lamb. Experiments with the pregnant ewe are curtailed by minimal transplacental transfer as well as limited access to the fetus. The EXTra-uterine Environment for Neonatal Development (EXTEND) model renders the isolated fetal lamb readily accessible and allows for direct fetal administration of sildenafil. METHODS: Five fetal lambs were placed on extracorporeal support in the EXTEND device and received continuous intravenous (IV) sildenafil (0.3-0.5-0.7 mg/kg/24hr) for a duration of one to seven days. Plasma sildenafil concentrations were sampled at regular intervals to establish the pharmacokinetic profile using population pharmacokinetic modeling. Serial Doppler ultrasound examination, continuous non-invasive hemodynamic monitoring and blood gas analysis were done to evaluate the pharmacodynamic effects and fetal response. FINDINGS: The target concentration range (47-500 ng/mL) was attained with all doses. Sildenafil induced an immediate and temporary reduction of pulmonary vascular resistance, mean arterial pressure and circuit flow, without change in fetal lactate levels and acid-base status. The duration of the systemic effects increased with the dose. INTERPRETATION: Immediate temporary pulmonary vascular and systemic hemodynamic changes induced by sildenafil were biochemically well tolerated by fetal lambs on extracorporeal support, with the 0.5 mg/kg/24 h dose balancing rapid attainment of target concentrations with short-lived systemic effects. RESEARCH IN CONTEXT: None. SEARCH STRATEGY BEFORE UNDERTAKING THE STUDY: A literature review was conducted searching online databases (Medline, Embase and Cochrane), using search terms: fetal OR prenatal OR antenatal AND sildenafil, without time-limit and excluding human studies. Where relevant, investigators were contacted in order to avoid duplication of work. EVIDENCE BEFORE THIS STUDY: Prenatal therapy with sildenafil, a phosphodiesterase-5 inhibitor with vasodilatory and anti-remodeling effects on vascular smooth muscle cells, has been considered for a variety of fetal conditions. One multicenter clinical trial investigating the benefit of sildenafil in severe intrauterine growth restriction (the STRIDER-trial) was halted early due to excess mortality in the sildenafil-exposed arm at one treatment site. Such findings demonstrate the importance of extensive preclinical safety assessment in relevant animal models. Transplacentally administered sildenafil leads to decreased pulmonary arterial muscularization, preventing or reducing the occurrence of pulmonary hypertension in rat and rabbit fetuses with diaphragmatic hernia (DH). Validation of these results in a higher and relevant animal model, e.g. fetal lambs, is the next step to advance clinical translation. We recently demonstrated that, in contrast to humans, transplacental transfer of sildenafil in sheep is minimal, precluding the in vivo study of fetal effects at target concentrations using the conventional pregnant ewe model. ADDED VALUE OF THIS STUDY: We therefore used the extracorporeal support model for fetal lambs, referred to as the EXTra-uterine Environment for Neonatal Development (EXTEND) system, bypassing placental and maternal metabolism, to investigate at what dose the target concentrations are reached, and what the fetal hemodynamic impact and response are. Fetal hemodynamic and metabolic tolerance to sildenafil are a crucial missing element on the road to clinical translation. This is therefore the first study investigating the pharmacokinetics, hemodynamic and biochemical effects of clinical-range concentrations of sildenafil in fetal lambs, free from placental and maternal interference. IMPLICATIONS OF ALL THE AVAILABLE EVIDENCE: We demonstrated self-limiting pulmonary vasodilation, a decrease of both systemic arterial pressures and circuit flows, induced by clinical range concentrations of sildenafil, without the development of fetal acidosis. This paves the way for further investigation of prenatal sildenafil in fetal lambs on extracorporeal support. A dose of 0.5 mg/kg/24 h offered the best trade-off between rapid achievement of target concentrations and shortest duration of systemic effects. This is also the first study using the EXTEND as a model for pharmacotherapy during pregnancy. ispartof: BIOMEDICINE & PHARMACOTHERAPY vol:143 ispartof: location:France status: published

Introduction: The surgically induced fetal lamb model is the most commonly used large animal model of myelomeningocele (MMC) but is subject to variation due to surgical technique during defect creation. Material and Methods: Thirty-one fetal lambs underwent creation of the MMC defect, followed by defect repair with either an extracellular matrix (ECM) patch (n = 10) or ECM seeded with placental mesenchymal stromal cells (n = 21). Postnatal hindlimb function was assessed using the Sheep Locomotor Rating (SLR) scale. Postmortem magnetic resonance imaging of the lumbar spine was used to measure the level and degree of spinal angulation, as well as cross-sectional area of remaining vertebral bone. Results: Median level of angulation was between the 2nd and 3rd lumbar vertebrae, with a median angle of 24.3 degrees (interquartile range 16.2–35.3). There was a negative correlation between angulation degree and SLR (r = –0.44, p = 0.013). Degree of angulation also negatively correlated with the normalized cross-sectional area of remaining vertebral bone (r = –0.75, p < 0.0001). Discussion: Surgical creation of fetal MMC leads to varying severity of spinal angulation in the ovine model, which affects postnatal functional outcomes. Postnatal assessment of spinal angulation aids in standardization of the surgical model of fetal MMC repair.

The possibility that prostaglandin E2 (PGE2) has a central action in the fetal lamb arises from studies by Ratter et al. and Robinson. The Prostaglandin E (PGE) release into the fetus may help to preserve fetal growth by decreasing nonessential fetal muscular activity, e.g., fetal breathing movements, thereby sparing glucose. PGE may also maintain a minimum glucose utilization by stimulating insulin release and enhancing pancreatic sensitivity to glucose. PGE2 concentrations in fetal plasma have been shown to increase during late gestation in sheep and this increase parallels the increase in fetal cortisol concentrations. The site of action of PGE2 has also been studied in newborn lambs in which plasma cortisol concentrations increased when PGE2 was infused into the brachiocephalic trunk, but not when infused into the descending aorta. Decreased placental perfusion, caused by placing a clamp on the common iliac artery, results in an increase in fetal cortisol concentrations, which may be secondary to increased fetal PGE2 concentrations.

We have developed a rechargeable fetal micropacemaker in order to treat severe fetal bradycardia with comorbid hydrops fetalis. The necessarily small form factor of the device, small patient population, and fetal anatomy put unique constraints on the design of the recharging system. To overcome these constraints, a custom high power field generator was built and the recharging process was controlled by utilizing pacing rate as a measure of battery state, a feature of the relaxation oscillator used to generate stimuli. The design and in vitro and in vivo verification of the recharging system is presented here, showing successful generation of recharging current in a fetal lamb model.

Recent fetal lamb data have suggested that the pulmonary trunk (PT) region displays a reservoir function and that a pharmacologically induced fall in pulmonary vascular resistance (PVR) increases and redistributes diastolic discharge from this central pulmonary reservoir toward the lungs, thereby producing a positive diastolic offset in the pulmonary arterial (PA) blood flow profile. As a similar offset in PA flow characteristically occurs after birth, this study tested the hypotheses that 1) central pulmonary reservoir discharge is both redistributed toward the lungs and increased in magnitude during the birth transition and 2) discharge from this reservoir constitutes a major component of increased PA diastolic blood flow after birth. Six anesthetized near-term fetal lambs were instrumented with PT, ductal and left PA transit-time flow probes, and aortic, PT and left atrial catheters. Hemodynamic data were recorded in fetuses and at regular intervals during 2-h mechanical ventilation following cesarean section delivery. Diastolic PA blood flow rose from near zero in fetuses to 468 ± 188 ml/min by 15 min ( P < 0.001). Central pulmonary reservoir discharge in fetuses (99 ± 44 ml/min) passed primarily right-to-left across the ductus. However, this reservoir discharge redistributed entirely to the lungs by 1 min after birth, and then doubled to a peak of 214 ± 167 ml/min at 15 min ( P < 0.001). Reservoir discharge subsequently stabilized at 151 ± 60 ml/min at 30–120 min, which comprised ∼50% of diastolic and ∼20% of mean PA blood flow. These findings suggest that enhanced diastolic central pulmonary reservoir discharge plays a major role in supporting an increased pulmonary perfusion after birth.

Birth asphyxia accounts for nearly one million deaths worldwide each year, and is one of the primary causes of early neonatal morbidity and mortality. Many aspects of the current neonatal resuscitation guidelines remain controversial given the difficulties in conducting randomized clinical trials owing to the infrequent and often unpredictable need for extensive resuscitation. Most studies on neonatal resuscitation stem from manikin models that fail to truly reflect physiologic changes or piglet models that have cleared their lung fluid and that have completed the transition from fetal to neonatal circulation. The present protocol provides a detailed step-by-step description on how to create a perinatal asphyxiated fetal lamb model. The proposed model has a transitioning circulation and fluid-filled lungs, which mimics human newborns following delivery, and is, therefore, an excellent animal model to study newborn physiology. An important limitation to lamb experiments is the higher associated cost.