Randomized Control Trial of Postnatal rhIGF-1/rhIGFBP-3 Replacement in Preterm Infants: Post-hoc Analysis of Its Effect on Brain Injury.

Background: Postnatal insulin-like growth factor-1 (IGF-1) replacement with recombinant human (rh)IGF-1 and IGF binding protein-3 (rhIGF-1/rhIGFBP-3) is being studied as a potential treatment to reduce comorbidities of prematurity. We have recently reported on a phase II, multicenter, randomized, controlled trial comparing postnatal rhIGF-1/rhIGFBP-3 replacement with standard of care (SOC) in extremely preterm infants (NCT01096784). Maximum severity of retinopathy of prematurity was the primary endpoint of the trial and presence of GMH-IVH/PHI one of the pre-specified secondary endpoints. Infants therefore received serial cranial ultrasound scans (CUS) between birth and term age. In this post-hoc analysis we present a detailed analysis of the CUS data of this trial and evaluate the effect of postnatal rhIGF-1/rhIGFBP-3 replacement on the incidence of different kinds of brain injury in extremely preterm infants. Methods: This report is an exploratory post-hoc analysis of a phase II trial in which infants <28 weeks gestational age were randomly allocated to rhIGF-1/rhIGFBP-3 or SOC. Serial cranial ultrasounds were performed between birth and term-equivalent age. Presence of germinal matrix hemorrhage and intraventricular hemorrhage (GMH-IVH), periventricular hemorrhagic infarction (PHI), post-hemorrhagic ventricular dilatation, and white matter injury (WMI) were scored by two independent masked readers. Results: The analysis included 117 infants; 58 received rhIGF-1/rhIGFBP-3 and 59 received SOC. A trend toward less grade II-III GMH-IVH and PHI was observed in treated infants vs. SOC. A subanalysis of infants without evidence of GMH-IVH at study entry (n = 104) showed reduced progression to GMH-IVH in treated infants (25.0% [13/52] vs. 40.4% [21/52]; not significant). No effects of rhIGF-1/rhIGFBP-3 on WMI were observed. Conclusion: The potential protective effect of rhIGF-1/rhIGFBP-3 on the occurrence of GMH-IVH/PHI appeared most pronounced in infants with no evidence of GMH-IVH at treatment start

The clinical presentation of preterm cerebellar haemorrhage

The objective of this study was to evaluate clinical symptoms and findings on cranial ultrasound (CUS) in preterm infants with cerebellar haemorrhage through retrospective analysis of all preterm infants with a postnatal CUS or MRI diagnosis of cerebellar haemorrhage admitted in a tertiary care centre between January 2002 and June 2009. Fifteen infants were identified; median gestational age was 25 2/7 weeks and median birth weight 730 g. We discerned six types of haemorrhage: subarachnoid (n=3), folial (n=1), lobar (n=9, of which 4 bilateral), giant lobar (n=1, including vermis) and contusional (n=1). Especially in infants with lobar cerebellar haemorrhage, CUS showed preceding or concurrent lateral ventricle dilatation, mostly without intraventricular haemorrhage (IVH). Thirteen infants suffered from notable, otherwise unexplained motor agitation in the days preceding the diagnosis. In conclusion, motor agitation may be a presenting symptom of cerebellar haemorrhage in preterm infants. Unexplained ventriculomegaly can be a first sign of cerebellar haemorrhage and should instigate sonographic exploration of the cerebellum

Metabolic engineering to simultaneously activate anthocyanin and proanthocyanidin biosynthetic pathways in Nicotiana spp

[EN] Proanthocyanidins (PAs), or condensed tannins, are powerful antioxidants that remove harmful free oxygen radicals from cells. To engineer the anthocyanin and proanthocyanidin biosynthetic pathways to de novo produce PAs in two Nicotiana species, we incorporated four transgenes to the plant chassis. We opted to perform a simultaneous transformation of the genes linked in a multigenic construct rather than classical breeding or retransformation approaches. We generated a GoldenBraid 2.0 multigenic construct containing two Antirrhinum majus transcription factors (AmRosea1 and AmDelila) to upregulate the anthocyanin pathway in combination with two Medicago truncatula genes (MtLAR and MtANR) to produce the enzymes that will derivate the biosynthetic pathway to PAs production. Transient and stable transformation of Nicotiana benthamiana and Nicotiana tabacum with the multigenic construct were respectively performed. Transient expression experiments in N. benthamiana showed the activation of the anthocyanin pathway producing a purple color in the agroinfiltrated leaves and also the effective production of 208.5 nmol (-) catechin/g FW and 228.5 nmol (-) epicatechin/g FW measured by the p-dimethylaminocinnamaldehyde (DMACA) method. The integration capacity of the four transgenes, their respective expression levels and their heritability in the second generation were analyzed in stably transformed N. tabacum plants. DMACA and phoroglucinolysis/HPLC-MS analyses corroborated the activation of both pathways and the effective production of PAs in T0 and T1 transgenic tobacco plants up to a maximum of 3.48 mg/g DW. The possible biotechnological applications of the GB2.0 multigenic approach in forage legumes to produce "bloatsafe" plants and to improve the efficiency of conversion of plant protein into animal protein (ruminal protein bypass) are discussed.This work was supported by grants BIO2012-39849-C02-01 and BIO2016-75485-R from the Spanish Ministry of Economy and Competitiveness (MINECO) (http://www.idi.mineco.gob.es/portal/site/MICINN) to LAC and a fellowship of the JAE-CSIC program to SF. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.Fresquet-Corrales, S.; Roque Mesa, EM.; Sarrión-Perdigones, A.; Rochina, M.; López-Gresa, MP.; Díaz-Mula, HM.; Belles Albert, JM.... (2017). Metabolic engineering to simultaneously activate anthocyanin and proanthocyanidin biosynthetic pathways in Nicotiana spp. PLoS ONE. 12(9). https://doi.org/10.1371/journal.pone.0184839Se018483912

Brain imaging in preterm infants at term equivalent age : Value and comparison of MRI and ultrasound

Cranial ultrasound (cUS) and magnetic resonance imaging (MRI) are the two most commonly used brain imaging techniques in preterm infants. cUS can be performed at the bedside and detects all major brain abnormalities (haemorrhages, infarctions, cysts, dilatation of the lateral ventricles) that are associated with severe neurodevelopmental disability. However, there are more subtle brain abnormalities that can be visualized with cUS, such as signs of brain atrophy and periventricular echodensities, for which the clinical relevance is less clear. Furthermore, there is no consensus how to quantify the lateral ventricular size on cUS, a procedure that is often necessary to monitor the progression of posthaemorrhagic ventricular dilatation or grade the extent of ventriculomegaly at term age. In comparison to MRI, cUs has been shown to be less sensitive for the detection of non-cystic white matter (WM) injury, the most common form of WM injury in preterm infants. Indeed, MRI at term age has been reported to be superior to cUS in detecting white matter (WM) abnormalities and predicting outcome in preterm infants. However, in that study cUS was performed during the first 6 weeks only and no late scan in parallel to MRI at term age was included. Furthermore MRI data from large population-based cohorts of extremely preterm infants are missing. Therefore, the aim of the present thesis was to study the relevance of subtle brain abnormalities on cUS (brain atrophy and periventricular echodensities) for neurodevelopmental outcome, to acquire and compare MRI and cUS data at term age in a population-based cohort of extremely preterm infants (< 27 weeks) and to determine which 2D linear cUS measurement of the lateral ventricular size is the best to estimate total ventricular volume using 3D MRI as the golden standard. Our main findings were, firstly, that sonographic signs of brain atrophy at term age, but not persistent periventricular echodensities, are related to neurodevelopmental outcome at 3 years of age, secondly that in the cohort of extremely preterm infants the incidence of brain abnormalities detected with MRI was unexpectedly low compared to previously published numbers from other regions, thirdly, that the 2D cUS measurements of the frontal horn and ventricular midbody correlate best to lateral ventricular volume, and finally that 40% of extremely preterm infants had a normal cUs at term age and that in these infants MRI at term age added no or only marginal clinically relevant information to the cUS result. In conclusion, our findings underline the important clinical role of cUs as a bedside imaging technique, but also highlight the potential of MRI as a complementary imaging tool in preterm infants